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BIOLOGY 4710: LIMNOLOGY

Course Outline

“Yes, as everyone knows, meditation and water are wedded forever … Why did the old Persians hold the

sea holy? Why did the Greeks give it a separate deity, and own meaning. And still deeper the meaning ofthat story of Narcissus, who because he could not grasp the tormenting, mild image he saw in thefountain, plunged into it and was drowned. But that same image, we ourselves see in all rivers andoceans. It is the image of the ungraspable phantom of life; and this is the key to it all. ”

- Moby Dick. Herman Melville

TEXT REQUIRED Limnology 3 rd ed., R. W. Wetzel. 2001. Academic Press. San Diego,CA.

LECTURE TOPIC

1. Importance of Limnology Chapter 12. Lake Formation Chapter 33. Light Chapter 54. Heat Budgets of Lakes Chapter 65. Water Movement Chapter 76. Oxygen Chapter 97. Carbonate Cycle Chapter 118. Chemical Constituents of Lakes Chapter 12,13,149. Phytoplankton and Zooplankton Ecology Chapter 1510. Productivity of Aquatic Systems Chapter 24, 25

LABSPlease refer to the Lab Manual.

Note: A weekend field trip is required for the second lab.

MARK BREAKDOWN Final (50%)Midterm (15%)Lab (35% - Includes a major write up for the field trip lab)

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BIOLOGY 4710- Limnology Laboratory Schedule Winter 2006

Lab: Description: Date:

1 Field Trip Preparation Jan .92 Map production (assignment) Jan. 16

3 Field I rip (Kingfisher Lake) Jan. 23

4 Water Quality Analysis – Alkalinity Jan. 30

5 & 6 Total Kjeldahl Nitrogen and Phosphorus Feb. 6(Combined Lab - only 1 report req'd)

7 Chlorophyll a Feb. 13

8 Lake Capacity/Heat Budgets Feb. 27

9 Water Quality Assessment Mar 6

10 Quality Assurance/Quality Control Mar 13

11 LC50 Quantal Test Mar. 20

12 TBA Mar. 27

LAB MARK DISTRIBUTION

Formal Report • May be completed to groups• Entitled "A Report on the Status of Kingfisher Lake"• Using data collected from labs 1,2,3,5&6 - 15%

Laboratories 4 and up - individual reports - 20%

Marking Key: Formal Report Labs Abstract 2 1Introduction 4 2Results 5 3Discussion/Conclusions 4 4Maps 3Presentation 2

Total 20 10

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FORMAL REPORT OUTLINE

"A Report on the Status of Kingfisher Lake"

A report should be completed on the status of Kingfisher lake, that includes a

brief Abstract which outlines the location of the lake, the time period that the study wascarried out over and a summary of the major findings, including the morphological datagained from the field trip preparation and water quality results obtained from the fieldtrip.

The report should also include all of the information requested in the laboratorymanual, but not details of the use of equipment. Equipment may be discussed if morethan one method was used to measure a lake parameter and a comparison was made.

The report should be comprehensive, but concise and probably not be morethan 10-15 pages, including maps. The reports can be a group report, which means

that the reports should be of a high standard, with carefully constructed maps with cleartitles, keys and scales.

Reports should follow the outline below:

Table of Contents

List of Figures/Tables (including maps)

Abstract - Brief summary of the report

Introduction - This section should include the purpose for carrying out limnologicalstudies of lakes and the design of the survey, i.e. which tests were carried out, whatdata was collected (where, when, and why). This section should also include adescription of the lake, including the location and access, and should be no more thantwo pages plus a map.

Results - This section should be the major section in the report and contain maps ordiagrams with some discussion. This section should be divided into MorphologicalAttributes, Water Quality and Physical Attributes.

Discussion/Conclusions - This should be a brief section which ties together all of the

data collected on Kingfisher Lake and makes some comment on the overall status, i.e. whether it has been highly developed or not and for what reason. Is the lake sufferingnoticeable signs of deterioration or are there any problems with water quality. Thissection should not need to be more than two pages.

References – This should follow the guidelines from the journal Limnology &Oceanography

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INTRODUCTION

The collection of accurate limnological data from water bodies is necessary foran understanding of the status of wetlands. Lake classification systems based ontrophic status, aid the management of these important resources. Limnological data

includes morphometric measurements; calculations of maximum length, width, area,volume; mean and relative depth; shore line and shore line development; measurementof physical characteristics such as temperature profiles, colour and turbidity; andchemical parameters such as pH, dissolved oxygen, alkalinity, concentrations ofinorganic and organic compounds and the composition and biomass of micro flora andfauna.

By the completion of this course, students will have the skills to:

• Plan a successful limnological data collection field trip.•

Identify and use suitable limnological field equipment.• Carry out suitable calculations to estimate morphometric parameters of lakes• Construct maps showing the major morphological parameters of a lake.• Identify suitable chemical analyses techniques to measure major inorganic and

organic fractions of water samples.• Estimate the trophic status of a lake from a collection of limnological data.

The practical section of this course will involve a field trip to a lake near Thunder Bay.The trophic status of the lake will be classified by assessment of morphological andwater quality parameters. This will also be an opportunity to practise water andsediment sample collection techniques.

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LABORATORY 1: Field Trip Preparation

Morphometry is defined as the methods of measuring and analyzing the physicaldimensions of a lake or stream (Coke 1994). In order to carry out limnological analysesof a lake, a detailed knowledge of the morphometry should be established, with

particular emphasis on the volume characteristics (Wetzel and Likens 1979). Criticalcomponents of detailed analyses of biological, chemical and physical properties of freshinclude depth analyses (including area measurements of sediments and water strata atvarious depths), volumes of strata and shoreline characteristics (Wetzel and Likens1979). For example, morphometric parameters are required to evaluate erosion,nutrient loading rates, chemical mass, heat content and thermal stability, biologicalproductivity, effectiveness of growth and other structural and functional components ofthe ecosystem (Wetzel and Likens 1979). Detailed knowledge of morphometry and flowcharacteristics of freshwater bodies is relied upon in management techniques, such asthe loading capacity for effluents and the selective removal of undesirable componentsof the biota (Wetzel and Likens 1979).

Detailed hydrographic maps of lakes and streams are often unavailable to thelimnologist. Those available from governments or other sources must be checked foraccuracy, as the morphometry of a body of fresh water changes with time. It istherefore of great importance for the student to have a general understanding of theconstruction of bathymetric (contour) maps and the computation of morphometryparameters (Wetzel and Likens 1979)

Before any field trip to survey the limnological parameters of a water body, a number ofpreparatory steps have to be completed:

1) Obtain the necessary maps, aerial photographs and gazetteer.

2) Locate and identify the water body.

3) Obtain access information for the water body.

4) Prepare suitable copies of a work (transect) map.

5) Calculate surface area and shoreline length including island shoreline.

6) Decide which limnological parameters are to be measured on-site and which areto be measured from collected samples.

7) Choose and familiarize yourself with the equipment needed to collect the dataand samples. Make sure the equipment is in working order and that there are

spare batteries for meters.

1.1. Identifying and Defining Water Body Location

1.1.1. Name and Location: Identifying and defining the location of a water bodymay require the use of the Gazetteer, Topographical maps, NationalWatershed Code Maps and Forest Resource Inventory Maps (FRI).

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The Gazetteer is the official source of information in Canada giving thename of the lake that corresponds to the geographical coordinates foundfrom a Topographical map The latest edition is 1974, with subsequentsupplements.Write down the name of the lake that you will be surveying.

The 1:50,000 Topographical Map is the most common and availablesource of information for identifying and distinguishing water bodies. Besure to check the scale of the map you are working from!

Write down the geographical position, general location and extent ofthe lake.

1.1.2. Elevation: The elevation is measured as the height above sea level of ftwater body. The elevation is often indicated on Topographical Maps,however smaller lakes are sometimes not and the elevation of these has

to be estimated from the surrounding land contours and known elevationsof connecting lakes. Elevation in feet has to be converted to metersGive the elevation of the lake.

1.1.3. Watershed Code: The Watershed Code Maps, number watersheds anddrainage systems m Canada. In Ontario, there are three main drainagesystems: The St. Lawrence (2), The Hudson Bay (4) and Lake Winnipeg(5). Each system has been divided into Principal Watersheds andWatershed Units An alpha-numeric code identifies each watershed unit,Identify the Watershed Unit Code for the study lake.

1.1.4. Access: Access describes the manner and direction one takes n reachinga water body. The access description should be short but precise.Give an access description for the study lake.

1.2. Map Preparation

Before a field trip, at least 4-5 maps should be produced for the following data maps:

1) Sounding transect map

2) Shoreline cruise map

3) Sampling map (indicating sampling/data collection locations with description of

what was collected)

4) Rough draft contour map

5) Final contour map*

6) One extra copy

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Lake Outline MapGazetteer name Jones Lake

Local name Jones Lake

MNR District Chatham

Township Elgin

Lat. and Long. 42°17’ - 81°19’

Area conversion factor 1.01

1.2.2. Lake Shoreline and Island Shoreline

Once the lake has been located on a FRI map the surface water area and perimeterof the lake and islands should be measured and recorded. These measurements

are done first because tracings and enlargements introduce error

The Map Measurer

This unit measures linear distance in centimetres or inches. Theindicator needle is set at zero by rotating the tracing wheel.Measure the shoreline by tracing the outline of the figure and thenread the result from the scale. Care must be taken so that theindicator needle travels In a positive direction. Also on a large lakethe needle may travel more than once around the circumference ofthe dial. Therefore, the operator should keep close watch as to thenumber of revolutions.

Use a start/finish line on the map to eliminate most of theseproblems.

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1.2.3. Measuring the water body area: The most accurate measurement of

lake surface area is by computer analysis of a digital image. Surface areacan be estimated from a dot grid overlay but for our purposes, we will usethe digital planimeter, which is more accurate.

Calculate the surface area of the study lake using the Digital Planimeter.Remember to set the planimeter to read in km 2.

Using the Digital Planimeter (Calculation of Lake Area)

1) Choose starting point on the map - centre the tracer lens on thispoint. Press ON.

2) Press UNIT key until km 2 is highlighted.

3) Enter the 1 : N scale value of your map by typing in N. i.e . If yourscale is 1:20000 then type in 20000 then press SCALE

4) Press START (the instrument will beep and zero).

5) Trace the lake outline in a clockwise direction until you reach yourstarting position. Once you are back at your starting position pressHOLD .

6) Move the tracer lens to the start position on any islands in the lakeand press HOLD .

7) Trace the island outline counter-clockwise to subtract the island

area from the lake area.

8) Press HOLD when you have finished tracing the island area.9) Press HOLD again.

10) Press END to store the first area calculation.

11) Repeat the above steps (8-10) two more times and record theareas. Remember to press END after each tracing.

12) Calculate the average area.

NOTE: For later calculations you will need to remember the following:

1 km 2 = 100 ha

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LABORATORY 2: Field Activities

Three exercises have to be completed on the field trip. The data that is collected will beused in a report to be handed in on Friday November 9. This report will includeinformation gathered from the first four laboratory sessions.

1) Echo sounding

2) Water Quality Data

3) Shoreline Cruise

2.1 Echo Sounding

The Contour Map

The Contour Map sometimes called abathymetric or

hydrographic chart is a graphic representation of the Lake

Bottom or lakebed as determined from depth soundings.

The Contour Map reveals the extent of shallows, the

degree of inclination of the lake bottom, the shoals and the

deep holes. To prepare the Contour Map, the various

depths of the water basin have to be measured by an echosounder.

PROFILE OF LAKE BOTTOM

The Echo Sounder

There are a number of models and types of echo sounders,

but all operate on the same principle - 'sonar'. This word is

derived from sound, navigation and ranging . Simply, sonar

measures and records time between the transmission of

Head Lake

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sound through water and the reception of an echo. A schematic diagram of the

principle is shown.

Furuno models are the more common instruments used in the program at this time,

although some Ferrograph models may be used.

Note: It is the responsibility of the operator to become familiar with the unit in use byreading the appropriate Instruction Manuals. Only some common features areexpressed in this manual.

FROM VIEW OF FG- 1/200 MARK-3 FURUNO

The Transducer must be completely submerged in water

to record properly. The face is normally secured to a

bracket mounted on the gunwale of the boat. Signals will

also penetrate unpainted aluminum and clear ice, if the

face of the transducer is submerged in water – alcohol

mixture for winter operation. Polyethylene bags hold the

liquid in these cases. For the purposes of our field trip,

the boats have an unpainted aluminum bottom, so the

transducer is placed in a polyethylene bag filled with water

The power supply for both the Furuno and Ferrograph

units is a 12V battery (wet or dry cell). Each unit

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normally has a voltmeter dial to indicate the power supply

strength. Furuno models should operate as efficiently

with eight transistor dry cell batteries as with twelve-volt

wet cells. You can expect problems though if you use

ordinary flashlight batteries,

The Ferrograph operates more efficiently on a 12-volt

wet cell or for short periods on two 6-volt dry cells wired

in series. The unscrewing of the scale-illumination bulb

from its holder will maintain the necessary voltage for

longer periods.

The signals received by the transducer are converted to

electrical energy, magnified by the amplifier, and

recorded on the electro-sensitive recording paper by the

rotating stylus.

The strength of the pulse that makes the mark on the

recording paper depends on the echo that produces it.Obviously, the strongest echo will be produced by the

true lakebed, but it is not unusual for multiple echoes of

the lakebed, as well as echoes of other objects,

including fish, to be recorded.

The resistance of the lake bottom to sound, and

consequently the number of multiple echoes, decreases

at this point with the degree of reflecting power in a

rock, sand or mud bottom. Objects with higher solidity

will generate fewer echoes.

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The resistance to sound also decreases with the inclination of the lakebed, i.e. a flat

lake bottom will give a better echo than a sloping one.

The degree of amplification influences both the information of multiple echo

recordings and the extent of the marking on the recording paper. Adjust the gain control

and refine the recording of the true lake bottom.

2.1.1 Echo Sounding ProceduresEcho sounding, i.e. running the sounding

transect line, is a flexible and schematic

procedure in collecting the required

depth data for the production of a

representative Contour Map.

Transects must be straight runs

from and to visible shoreline features

which can be identified on the field map,

i.e. projections, centre of indentations,

landmarks, etc.

Each transect must be run at a constant outboard motor speed. Speeds may

vary from transect to transect, but never during the transect. If variation occurs, return

to the starting point and restart the sounding run.

Line number, direction of run and distance from shoreline(s) of the sounding

recording terminals must be indicated on the transect map and on the tape. This

eliminates any doubt in the interpretation of the sounding tape.

It is important to map the extent of all reefs and shoals as these are favourite

feeding and/or spawning sites. This can be done by running extra sounding lines in

deeper water or sounding with a weighted hand line in shallower water that is

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hazardous for echo sounding. A paddle with marked graduations is a handy tool for

spot sounding in shallow water, i.e. behind small islands. Mark the depths directly on

the 'transect map'. If the starting and finishing distance from shore is constant, then this

can be marked on the map in some prominent position. This remedies needless

duplication of effort. Otherwise, mark distance to shore at the start and stop of each

transect on the map.

Transect line numbers on the transect map correspond with numbers marked on

the tape. Note the Date, Lake Name, District, Latitude and Longitude, and Sounder

Tape Type (A or B) on each sounding tape, if there is more than one.

2.1.2 The ‘E’ Line

The ‘E’ Line is an exploratory line and will always be the initial line run. If the

basin shape is simple, i.e. reasonably oval or rectangular without indentations, then one

‘E’ Line will normally suffice. If the basin shape is more complex, then more than one

‘E’ Line is required. ‘E’ Lines will run the length of the basin or segment, midway

between the shorelines. It is not required to be a straight line. It will be shown as

illustrated on the transect map.

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The running of the number 1 sounding line will commence at or near the finish

terminal of the ‘E’ Line. ‘E’ Lines dictate sounding density: The more irregular the

bottom profile, the greater the number of transect lines required. It is mandatory that

the lake be adequately covered by sounding lines, but sampling too many wastes time

and energy.

2.1.3 Transects

Transects are always run in a zigzag fashion across the breadth of the basin(s).

Eyeball the ‘E’ Line to determine the number and location of transects, so that

underwater formations can be isolated.

Crossing transect lines should be avoided where possible, as this often causes

confusion when drawing the Contour Map. Also, transect lines should always be kept

as short as possible and perpendicular to shore. In this way, there is less chance of

going off course and a constant speed can be maintained.

a) On one of your maps, decide where your ‘E’ line is going to be and draw it in. Next,

decide on your transect lines and mark them on the same map, starting at number one.

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2.2 Water Quality Data

When you have read the section on water quality, familiarize yourself with the

equipment that you will be using on the field trip. Make sure that you know how to set

up the water samplers and obtain a sample from them. Know how to use the secchi

disc and the min/max thermometer. Also, make sure that you are familiar with the

meters that will be used on the field trip.

After completion of the echo sounding (at least partially), the water chemistry

tests are carried out. These tests combine measurements of physical and chemical

parameters. However, since they are done at the concurrently, they will be treated as

one unit in this manual. A complete chemistry test consists of all the following

measurements:

1) Secchi disc

2) Cloud cover

3) Water surface

4) Water colour

5) Air temperature

8) Water temperature profile7) Dissolved oxygen

8) pH

9) Alkalinity

10) Conductivity

11) Cell temperature

12) Time of day

For the purposes of our field trip, we will attempt all of the above tests except for

alkalinity (see Section 2.2.1 to 2.2.8 ). Water quality data will be obtained after transects

are completed. This testing will be done at the deepest part of the lake. Each group of

students will obtain water samples for further analysis. Water temperature profile,

dissolved O 2, pH and conductivity will be done as one large group.

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Choosing Chemistry Stations

The selection of water chemistry stations is very important. The station should

reflect as much as possible the situation in the lake at the time of the survey and if

future sampling occurs, that this information can be used as time sequence data. Too

few stations tell little about the lake, while too many can provide little additional

information and contribute to wasted time. Each station is assigned a number; the

station at the deepest part of the lake should be labelled number one, regardless of the

sampling order.

Basin Differentiation

Surveyors should keep in mind that the inventory program must describe for thefast time many characteristics of the water body. In order to undertake this, the

following criteria are recommended as guidelines in establishing chemistry stations:

1) The number one chemistry station should be conducted in the deepest

basin of the water body. If the echo sounding tapes indicate more than one

basin of equal depth (with +- 5 metres), then sample each basin at the deepest

part.

2) If there is a basin within proximity to an inlet(s) and/or an outlet(s), a

sample station should be set up. However, do not sample where there is

movement of water, since this data will reflect the inlet characteristics and not

those of the lake. In addition, if the discharge rate is less than 0.5m 3 /s, a station

is not required.

3) If a basin has a depth 75% of the deepest basin maximum depth, a

sample station should be set up.

4) If there are bays that are isolated from the main body of water, it is

advisable to set up a station.

5) As a rule of thumb, there should be at least one chemistry station- for

every 250 hectares of water surface-area.

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LABORATORY 3: Map Preparation

After completing the field activities, you are responsible for the following:

3.1 Physical Features and Contours: Construct maps of the physical features and

contours. The following instructions will show you how to produce your maps

3.2 Morphometric Calculations: Perform the calculations listed below (instructionsfor completion of calculations are given in the following pages).

1) Areas of the surface and each contour interval at depth z (contour areas for eachcontour)

2) Lake Volume

3) Mean Depth

4) Shoreline Development Factor

5) Maximum Depth

3.1 PRODUCING THE CONTOUR MAPThe contour map is prepared by transcribing the depths from the echo sounding tapes

to the corresponding transact map. This can be done using one of two methods. Thepreferred method is using proportional dividers. Due to the lengths of the soundingtapes, we will only do a couple of transects using the dividers. A ratio method

Proportional Dividers

Short Tips

Long Tips

Setting Knob

AdjustingKnob

Fulcrum

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(described further on) will be used for the remaining transects. It will not be as accuratebut will serve our purposes. Proportional dividers are instruments made of crossed double pointed metal arms.They are made in such a way (as the name suggests) that they can be adjusted tomeasure ratios or proportions.

The tips on each end of the arms are of different lengths. One end has long tips andthe other end has short tips (See diagram.) The fulcrum also can be adjusted andmoved towards either end by loosening the adjusting knob and turning the setting knob.The arms must he closed together when moving the fulcrum.

When transcribing depths from the tape to the map one transect is covered at a time,the dividers must then be adjusted so that the ends with the longer tips extend thelength of the longer line and at the same time the ends with the shorter tips extend tothe length of the shorter line. (See diagram).

NOTE: Adjusting the dividers to the correct position can be tricky.See the following instructions.

Adjusting the DividersTo find the right position for the fulcrum of the dividers, there are two methods:

1) Measure accurately the length of the lines and calculate the ratio. The dividers canthen be set using the scale on the instrument and by following the supplied formula;each model type has a different formula. This method involves time-consumingmeasurements and calculations and therefore is not recommended.

2) Set the dividers by trial and error. This method is quick and involves nomeasurements or calculations

Trial and Error Method

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a) Decide which line is longest (i.e. either the transact line on the map or the length of

the sounding run on the tape).

b) Place the dividers with the longer tips on the ends of the longer line (usually the

transect line on the map) as in the previous diagram.

c) With the dividers in the same position, place the dividers with the shorter tips onto the

ends of the shorter line (usually the sounding tape).

d) If the distance between the shorter tips is less than the length of the line then

proceed with step e . If the distance between the shorter tips is greater than the length

of the line then proceed with step f.

e) Close the dividers and adjust the fulcrum by moving it towards the long points. (See

the following diagram) Then repeat steps b , c , d , and a until a correct setting is

obtained. Close the dividers and adjust the fulcrum by moving it towards the shorter

points.

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f) Close the dividers and adjust the fulcrum by moving it towards the shorter points (See

the following diagram). Repeat steps b , c , d , e and f until the correct setting is

obtained.

Dividers Closed

Adjusting the Fulcrum

1) Loosen the adjusting knob.

2) Turn the setting knob.

Note: Usually the correct setting can be obtained in 2 - 3 tries. Speed will increase withexperience.In addition, if the same general boat speed has been maintained for each soundingtransect, then very little adjustment of the proportional dividers will be necessary foreach transect, as the ratio will be consistent.

When lines are too long for the dividers, divide the lines into equal sections and doseparately.

Transcribing Depths

Once the proportional dividers have been adjusted the depths can be transcribed from

the sounding tape to the transect map.

Note: Adjust the dividers for each transect (run).

1) If the length of the sounding run on the tape is shorter than the map transect line,

then place the shorter ends of the dividers on the tape. I.e. for the 2 m depth place one

tip on the start line of the run at the 2 m level. The other tip is placed on the 2 m line at

the point that the bottom echo reaches 2 m in depth. See diagram opposite.

Fulcrum

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2) Remove the dividers, making sure that the setting is not disturbed

3) Place the long points on the corresponding transect line. One long point should be at

the beginning of the run while the other will cut the line as in the diagram. This point is

then marked and the appropriate depth is marked. E.g. 2 m.

4) This technique is repeated for all the depths along the transect line.

5) Each transect line is transposed as described above.

Note: Be sure that the depths are plotted from the start of the run on the transect map.

6) For both the beginning and end of transect lines, allow for distance from shore.

Ratio Method

This method will be used for the rest of the transect lines from your sounding

runs (This is a time saving measure). Usually, we use this method when the transect

lines are too large to use the proportional dividers. Long transects may be broken up

for use with the dividers, but this consumes too much time.

To calculate the ratio:

1) Measure the length of a transect line on your map.

2) Measure the length of the corresponding transect on your sounding tape.

3) Divide 1) by 2). This produces your ratio, i.e.

Map distance (1 transact line) = RatioTape distance (1 transect line)

4) Multiply the ratio by the distance from the edge at each depth interval (4 m, 8 m, 12

m, etc.)

E.g. ratio x 4 m

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Required Contour Depths

The more depth contours on a map, the more accurate the calculations for

volume and mean depth. However, discretion is important when plotting depths; the

contours could become too closely spaced, blurred and meaningless. On the other

hand, too few lines will give a less than accurate description.

As a rule of thumb, the prescribed contours to be drawn (in metres) are 1, 2, 4, 6,

8 and thereafter by multiples of 2. If the lake is very deep and has a steeply sloping

lakebed, multiples of 4 m can be used. i.e . 2, 4, 8, 12, etc

Note: Do not use odd numbers, i.e. 1, 3, 5, 7, 9, etc.

In shallow lakes of less than 10 m in depth, contour intervals of 1 m may be more

appropriate, i.e. 1, 2, 3, 4, etc.

Drawing the Contours

Once all the depths have been transcribed from the sounding tapes to the

transect map, then join the points of equal depth on the map.

Join up the depths as accurately as possible. Sometimes when steep drop-offs

occur, it may be necessary to run contours into each other or into the shoreline. This is

quite common when drawing on the one-metre contour. See example contour map

illustrated.

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The Final Contour Map

Once the initial contour map is completed, it is traced to produce the final map which willnot show the transect lines. A sample map appears at the end of this section.

3.2 Morphometric Calculations

3.2.1. Measuring Contour Areas

Measure the surface area from the contour map using the Digital Planimeter (for

instructions on the use of the digital planimeter, see Laboratory 1 ). Trace the

contours at each depth level.

Note: When measuring the contour areas, measure all of the water area within

the appropriate contour. I.e. The 2 m contour area includes all the water within

the 2 m including the 4, 6, 8, etc.

You will need the following conversions:

1 ha = 10 4 m 2

1 km 2 = 100 ha

3.2.2. Volume Calculation

INITIAL CONTOUR MAP

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The volume of the basin is the sum of the strata volumes at successive

depths, from the surface to the point of maximum depth (Wetzel and Likens,

1979). To calculate the volume of a lake, measure the surface area of each

contour. Then, use the formula for Total Lake Volume for the calculation.

3.2.3. Mean Depth

Mean depth, z in metres, is calculated by dividing the volume ( V) of the

lake by the area ( A).

(Surface area of the lake at zero depth).

z = V(10 4m3)A(ha)

3.2.4. Shoreline Development Factor (S. D.F.)

Shoreline Development Factor describes the irregularity of the shoreline

by relating shoreline length (perimeter) to the length of the circumference

of a circle with the same area as the lake – Island shoreline is not part of

this calculation.

E.g. the S.D.F. of a perfectly circular lake is 1.0. Therefore, S.D.F. is

always more than 1.0.

As the length of the shoreline becomes more irregular, the shoreline

development deviates more and more from the minimum S.D.F. value of

one. Only a few lakes approach this circular shape. E.g. Crater Lake in

Oregon, and a few Kettle lakes.

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Many sub circular and elliptical lakes have a S.D.F. of about 2. Lakes of

flooded river valleys have much larger S.D.F. values. Shoreline

development is of interest because it reflects the potential for development

of littoral communities, which are usually highly productive.

S.D.F. = P .

2√π A

P = shoreline perimeter (km)

A = area (km 2)

π = 3.14

3.2.5. Maximum Depth

Maximum depth in metres is the greatest depth recorded for the whole

lake .

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LABORATORY 4: WATER QUALITY ANALYSIS

Chemical Analysis

Precipitation falling upon the surface of the earth, as either rain or snow, containsa variety of dissolved gases. Additionally, incorporation of aerosols or dust particlesoccurs along the way. Therefore, by the time the water reaches the earth's surface, it isno longer pure. As it flows over, or penetrates into plants and soil, water dissolves moregases. Notably, carbon dioxide and various mineral substances with which it has comeinto contact. CO 2 dissolves in water, forming carbonic acid and lowering the pH:

H2O + CO 2 ↔ H2CO 3

Most minerals are only slightly soluble in water. For example, limestone (calciumcarbonate, CaCO 3) is slightly soluble in pure water. However, limestone is more solublein water containing carbonic acid; the conversion of CaCO 3 to calcium bicarbonateallows more limestone to dissolve:

CaCO 3 + H 2CO 3 ↔ Ca(HCO 3)2 (Insoluble) (Soluble)

The dissolved bicarbonate has a marked effect on the chemical properties of thewater. Ca(HCO 3)2, for example, dissociates into Ca 2

+ and HCO 3 –

ions. The HCO 3 –

ions react with H + (which originated from the natural dissociation of H 2O) to formcarbonic acid (H 2CO 3). The H 2CO 3, in turn, dissociates into soluble CO 2, which is oftenin equilibrium with CO 2 from the air and H 2O. Therefore, the bicarbonate changes thepH of the water, increases the alkalinity of the water, and imparts hardness to the water.The amount of dissolved salts in water is important to the maintenance of life and is animportant factor in the treatment of the water for domestic and industrial use.

In addition to bicarbonates, carbonates, and hydroxides, other moderatelysoluble minerals are silica, chlorides, sulfates, and nitrates of calcium, magnesium,sodium and potassium. With increased emission of sulfur dioxide (SO 2) into theatmosphere from industrial activities, SO 4

– is becoming the dominant anion in

precipitation in large geographical areas. Consequently, serious alteration of thegeochemical relationships can occur.

In natural waters, inorganic carbon as dissolved CO 2 and HCO 3 – is the primarycarbon source for photosynthesis by algae and larger aquatic plants. In addition torespiratory production of CO 2 by most organisms, influxes of CO 2 and HCO 3

– from

incoming water and from the atmosphere balance this utilization. The amounts ofbioavailable inorganic carbon are adequate in most natural fresh waters; only underspecial conditions, such as soft waters and intensely productive situations, doesinorganic carbon become a limiting factor to photosynthesis.

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Natural waters exhibit wide variations in relative acidity and alkalinity, not only in

pH values, but also in buffering capacity. The concentrations of compounds and theratios of one to another determine the observed pH and the efficiency of buffering of agiven body of water. The lethal effects of most acids appear when pH < 5.0 and of most

bases near pH 9.5, although the tolerances of many organisms are considerably morerestrictive. Therefore, the capacity of natural waters to resist changes in pH is veryimportant to the maintenance of life.

Alkalinity of fresh waters refers to the measure of the capacity of water toneutralize acids. The main contributors to alkalinity in water are bicarbonates,carbonates, and hydroxides; and less frequently by borate, silicate, and phosphate.Since CO 2 is relatively abundant in both gaseous and dissolved form, and bicarbonatesand carbonates are common in primary minerals over wide areas of the earth,carbonate anions usually dominate the buffering system of fresh waters. Directcontributions to alkalinity by hydroxides are rare in nature, except in very nutrient-poor

waters.The interrelationships between carbon dioxide and the other major components ofalkalinity are as follows:

Free CO 2 in the air is often in equilibrium with dissolved CO 2 in the water. Thisequilibrium, together with the other equilibria taking place in the water, isrepresented by the following equation:

CO 2 (air) ↔ CO 2 (dissolved) + H 2O ↔ H2CO 3 ↔ H+ + HC0 3

– ↔ H+ + CO 32

–

The concentration of CO 2 in the atmosphere averages about 0.03 percent, butvaries with location. Photosynthesis and respiration by aquatic organisms substantiallyinfluences the quantity of CO 2 in water at any given time and place. After equilibrium isestablished in the water, the resulting condition is exemplified by the equations:

CO 2 + 2H 2O ↔ HCO 3 – + H 3O+ (1) HCO 3

– + H 2O ↔ H2CO 3 + OH – (2)

H2CO 3 ↔ H2O + CO 2 (3)

The hydroxyl ions (OH –

) formed (equations 1 and 2) show why waters with highcarbonate content are alkaline. Acid (source of H +) must be added to bring the water tothe point of neutrality, i.e. where equal quantities of hydronium and hydroxyl ions arepresent.

The equilibria shown by equations 1, 2, and 3 explain the buffering capacity ofalkaline waters. That is, the water tends to resist change in pH as long as these

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equilibria are in existence. Addition of H + (Acid) reacts with the OH –

formed in equation2; in response, reaction of carbonate with water forms more OH

– , as long as excess

carbonate is present. Therefore, the pH remains unchanged until the available supplyof carbonate and bicarbonate is exhausted. Similarly, addition of OH - initiates thefollowing reaction:

HCO 3 –

+ OH –

↔ CO 3 – + H 2O (4)

The terms alkalinity, total alkalinity, alkaline reserve, titratable base, or acid-binding capacity are frequently used to express the total quantity of base, in equilibriumwith carbonate or bicarbonate, that can be determined by titration with a strong acid.Alkalinity is the equivalent concentration of titratable base and is determined by titrationwith a standard solution of a strong acid, e.g. , 0.02N H 2SO 4, to certain equivalencepoints as given by indicator solutions. The indicator phenolphthalein is commonly usedfor the measurement of that portion of the alkalinity contributed by hydroxyl and

carbonate ions, while an indicator responding in the pH range below 5 is used tomeasure the alkalinity contributed by bicarbonate (See figure below).

To facilitate calculations, all alkalinity sources are expressed as eithermilliequivalents per Litre or milligrams calcium carbonate per Litre. The latter is termedTotal Alkalinity as calcium carbonate . From a practical standpoint, this method ofexpressing alkalinity is satisfactory and is used extensively in limnology.

Relation between pH and the relative proportions of inorganic carbonspecies of CO 2, HCO 3

– , and CO 32 – in solution (From Wetzel, 1975)

The measurement of chemical variables is the basis for most water qualitymonitoring programs. Water quality can have significant social, economic andenvironmental implications because the quality of water determines its usefulness. Theprominent chemical variables in aquatic wetlands are pH, dissolved oxygen,conductivity, alkalinity and inorganic nutrients.

pH

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The pH of a water body is a measure of the hydronium ions present in solution. Inlakes, the pH is affected by the amount of carbon dioxide dissolved in solution,biological activity and the pH of incoming waters in the catchments. pH is usuallymeasured directly in the field because it is readily altered by biologically activity andtemperature.

Conductivity

Conductivity is a measure of the electrical resistivity of a solution. Conductivity dependson the presence and concentration of ions, particularly the simple inorganic ions Na +,Ca 2+, Mg 2+, K+, CI -, HCO -, and sulfur based oxoanions (SO xy). Higher concentrations ofions mean lower electrical resistivity and therefore, higher conductivity. Additionally,conductivity can measure salinity, because salinity is the total concentration of ions insolution (including the ions mentioned previously).

Since conductivity is generally stable, with respect to biological activity, then

measurements can be made directly in the field or with samples in the laboratory later.Measurements with a portable meter give readings in micro-Siemens per centimetre.

Total Dissolved Solids (TDS)

TDS is a measure of organic and inorganic materials in water and is related to salinity.TDS is determined from the weight of remaining material following evaporation, at adefined temperature – usually 180 °C –, of a filtered sample of known volume. Inaddition, conductivity can estimate TDS via multiplication by a conversion factor – usually 0.6.

Dissolved oxygen

The distribution of O 2 in natural waters provides a measure of organic production anddecomposition, and is a basis for most methods of measuring primary productivity. TheWinkler titration method is routinely used for measuring oxygen concentration. It is asimple oxidation-reduction reaction involving manganese, iodine and thiosulfate.

AlkalinityAlkalinity is the common method for determining carbonate content and the bufferingcapacity of water. Additionally, alkalinity is an index to the rock contents within adrainage basin and the degree to which they are weathered. Limestone present indrainage basins will increase the carbonate content through dissolution of CaCO 3 andtherefore increase the alkalinity.

To determine alkalinity, a sample is titrated with a standardized acid – usually sulfuric –to the equivalence point of appropriate indicators (phenolphthalein and bromcresolgreen - methyl red).

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In environmental literature, two kinds of alkalinity are usually reported: phenolphthaleinalkalinity and total alkalinity. Phenolphthalein alkalinity (P) measures the bufferingaction of bases as strong as, or stronger than, the carbonate ion (CO 32

– ).

Phenolphthalein alkalinity exists when the pH is greater than 8.3. Whenphenolphthalein is used as the titration indicator, the color of the water sample will

change from pink to colorless when the pH of the sample has decreased to 8.3. Thisrepresents all of the hydroxide alkalinity, 1/2 of the carbonate alkalinity, and 1/3 of thephosphate and any other alkali producing material present in the sample above a pH of8.3. There is usually no hydroxide alkalinity in water of pH less than 9.2. If the samplewater is initially below pH 8.3, the phenolphthalein alkalinity is zero.Sample reactions that occur during the titration include:

OH –

+ H + (from sulfuric acid) → H2OCO 32

– + H+ → HCO 3

–

Adding bromcresol green-methyl red indicator to the water sample will turn it a blue-green color. Bromcresol green-methyl red measures the buffering action of bases asstrong as, or stronger than, the bicarbonate ion (HCO 3

– ). Adding acid to the sample will

change the colour to pinkish-purple when a pH of 4.3 is reached. The following reactiontakes place during Bromcresol green-methyl red alkalinity determination:

HCO 3 –

+ H+ (from sulfuric acid) → CO 2 + H 2O

Total (T) alkalinity is the sum of the phenolphthalein alkalinity and the bromcresol green-methyl red alkalinity. This represents all of the hydroxide, all of the carbonate, and 2/3of the phosphate and other alkali producing material present in the sample above a pHof 4.3. The pH of natural waters is normally less than 8.3 so there is no P alkalinity.They also do not normally have a pH below 4.3 so they do not contain strong mineralacids.

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Samples were collected from a pond, creek and a well, all in the same geographicallocation. Determine the alkalinity of each sample using the method below. Replicatethe analysis for each sample and evaluate the results. In your discussion, be sure tomake note of any differences in the results for each sampling source and betweenreplicates and possible reasons for those differences. What are the sources of error?

APPARATUS: 50 mL burette stand and holder250 mL Erlenmeyer flask funnelPasteur pipette

REAGENTS:0.02 N H2SO 4 Phenolphthalein indicatorBromcresol green-methyl red indicator

METHOD:

1) Assemble the titration apparatus Rinse the burette with a few mL of 0.02NSulfuric acid and then fill the burette to the 50 mL mark. Use caution anda funnel.

2) Pour 50 mL of the sample into the Erlenmeyer flask (250 mL). Take apiece of white paper and place it underneath the Erlenmeyer.

3) Add 4 to 5 drops of phenolphthalein indicator to the sample. If a pinkcolour appears, add the sulfuric acid slowly until the pink colourdisappears upon swirling, Note the volume of acid used, this volume isrequired to calculate the phenolphthalein alkalinity.

4) Then add 3 to 4 drops of bromcresol green-methyl red indicator to thesame sample. Slowly continue to add the sulfuric acid to the appropriateequivalence point (faint pink). The colour change will be green to clear tofaint pink. Remember to swirl the flask after each addition of acid andwhen you are approaching the endpoint, add the acid drop-wise. Note thetotal volume of acid used; this is required for the calculations of totalalkalinity.

NOTE: Faint pink means barely visible to the naked eye

5) Replicate the analysis for the sample. Compare the results.

6) Repeat this procedure for all three samples. (Six analyses in total)

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7) Calculate the phenolphthalein and total alkalinity for the water samplesusing the following equations:

Phenolphthalein alkalinity as mg/L CaCO 3

Volume of acid used to first endpoint (mL) x (normality of acid x 50000)Volume of sample (mL)

Total alkalinity as mg/L CaCO 3

Total volume of acid used to second endpoint (mL) x (normality of acid x 50000)Volume of sample (mL)

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LABORATORY 5: TOTAL KJELDAHL NITROGEN

Compounds of nitrogen and phosphorus are major cellular components oforganisms. Since the availability of these elements may be less than biologicaldemand, then environmental sources can regulate or limit the production of organisms

in freshwater ecosystems.

Concentrations of nitrogen and phosphorus compounds are highly dynamicbecause they may be utilized, stored, transformed and excreted rapidly and repeatedlyby various aquatic organisms. Measurements of these elements are complicated by thechemical form in which they occur. Ionic concentrations are often very low, requiringcare in collection and analysis of water samples, to avoid contamination.

Total Kjeldahl Nitrogen

Total Kjeldahl Nitrogen (TKN) includes the organic nitrogen compounds plus theammonia fraction but does not include NO 3 and NO 2 present in a water sample. Theprocedure has been developed by L.U.E.L. and is as follows:

WTKN: Total Kjeldahl NitrogenDetermination by the difference of WTOTN and WNOXWTKN = WTOTN - WNOX

WTOTN: Total Nitrogen (UV digestible)Using the Skalar autoanalyzer system, the sample is mixed with a potassiumperoxodisulfate/sodium hydroxide solution and heated to 90°C. The solution is thenmixed with a borax buffer and all nitrogen species are converted by UV radiation tonitrate. Colorimetric determination follows WNOX. This method accounts for nitrogen inthe form of nitrate, nitrite and a variety of other forms.

WNOX: Nitrate and NitriteUsing the Skalar autoanalyzer system, the nitrate is reduced to nitrite in a commerciallypacked cadmium column treated with copper sulfate. All nitrite is then determined bydiazotizing with sulfanilamide and coupling with N-(1-naphthyl)-ethylenediaminedihydrochloride to form a highly coloured azo dye (Griess reaction). The absorption ismeasured at 540 nm.

The environmental lab will process the Kingfisher lake samples using the Skalar

autoanalyzer. You will be provided with the WTOTN and WNOX values and will have tocalculate the value for Kjeldahl Nitrogen for your report.

Discuss TKN to an appropriate extent in your report and be sure to include a discussionon the reasons for variability in your results (if any) between the three depths sampled.

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LABORATORY 6: TOTAL PHOSPHORUS

Phosphorus plays a major role in metabolism in the biosphere and is of significantinterest ecologically. There is a relatively rich supply of other major nutritional andstructural components of the biota (C, N, O, and S); however, phosphorus is the leastabundant of the components and consequently limits biological productivity. Unlikenitrogen, phosphorus does not have a large immediate storage reservoir, theatmosphere.

Phosphorus occurs predominantly as phosphates in both natural waters andwastewaters. There are three classifications of phosphates: condensed phosphates,ortho-phosphates and organic phosphates. Phosphates occur in solution, in adsorbedto particles or detritus, or within the bodies of organisms.

These different forms of phosphates arise from a variety of sources:

A) Condensed Phosphates Small quantities periodically added to water suppliesduring treatment

Larger quantities added when the water fromlaundering or cleaning (major constituents of manycommercial cleaning compounds)

B) Ortho-phosphates Applied agricultural or residential fertilizers,transported to surface waters via storm runoff andspring melt

C) Organic Phosphates Formed primarily by biological processes, form a partof sewage through body wastes and food residue

May also be formed from orthophosphates inbiological treatment processes or by receiving waterbiota

The stimulated growth of photosynthetic micro- and macro-organisms to nuisancequantities, in phosphate limited aquatic systems, stems from the discharge of raw ortreated wastewater, agricultural drainage or industrial waste into the system.

Sample Digestion

Phosphorus analysis is generally a two-step procedure:

1) Conversion of the phosphorus to dissolved orthophosphate

2) Colorimetric determination of dissolved orthophosphate

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Samples collected from Kingfisher Lake have undergone a sulfuric acid digestionprocedure prior to the lab. This digestion oxidizes organic matter, to releasephosphorus as orthophosphate. The digested samples are ready for total phosphorusanalysis using a colorimetric determination of dissolved orthophosphate. The intensity

of colour, indicated by absorbance (ABS) values, is proportional to the amount ofphosphate present in a sample. A standard curve and regression equation is calculatedby performing a linear regression analysis, on the ABS values obtained for knownstandards using a spectrophotometer. The regression equation determines unknownconcentrations of phosphorus in your sample as a function of the sample ABS values.

Determination of Phosphorus (Phosphorus in water by the Ascorbic AcidMethod)

To create the standard curve, four phosphorus standards are used.

Standard Values in mg/L

S0 = 0.0S1 = 0.020S2 = 0.040S3 = 0.060C

NOTE: Safety glasses MUST be worn when working with acids (the CombinedReagent includes acid) .

Combined Reagent:

30 ml Ammonium Molybdate solution100 mL 5N H2SO 4 60 ml Ascorbic Acid Solution10 ml Potassium Antimony Tartrate

To each of the Standard Solutions and digested samples, add 4.0 mL of CombinedReagent using the pipette provided. Cover the test tube with Parafilm and mix gently,by inversion. It is important to avoid vigorous mixing of your samples, becausedissolved gas interferes with the absorbance measurement. After 10 minutes, butwithin 2 hours, pipette the solution into a cuvette and read the absorbances.

Absorbance at 880 nm (A 880 ) is measured on the CARY 50 Spectrophotometer with 10mm cuvettes. This instrument is a single beam spectrophotometer that measures thedifference between a reference solution and a sample solution.

The CARY is zeroed using a 0.0 mg P/L reference solution of degassed DDW. Oncethe CARY has been zeroed, the standards and samples can be measured.

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Measure the Absorbance (A 880 ) of each standard, from lowest to highest concentration.Rinse the cuvette with degassed DDW between each sample and discard the rinsate inthe waste container. Be sure to touch only the frosted sides of the cuvettes.Fingerprints will alter your readings. Wipe the clear sides of the cuvettes with Kim-wipes only. Continue the analytical run with your samples.

Once you have obtained your A 880 values for the standards and samples, perform alinear regression analysis to calculate the concentration of phosphorus, as a function ofthe absorbance units.

To create a standard curve, use the concentration of phosphorus as your X-axis(independent variable) and the A 880 as your Y-axis (dependent variable); plot the datafor the phosphorus standards. Once you have plotted your regression line, you candetermine the concentration of Total Phosphorus for your samples. By plotting yourdata you can then estimate on your regression line what the concentrations should befor your samples or simply, enter your values into the least square's line that you will

have calculated and solve for X (TP conc. for each sample).TP = (ABS - intercept)/slope

Use all the obtained data. Averaging the results from each group for the three thermallayers will give results that are more accurate. Include your regression line andcalculated Total Phosphorus results as an appendix to your report. The results from theSkalar Autoanalyzer should be used for your report as these results will be the mostaccurate. The method used on Skalar is similar to the procedure used in today's lab.This procedure has been developed by LULL and is as follows:

WP04: Reactive Phosphorus (P04-P)For the determination of dissolved reactive phosphorus, filtration of the sample througha 0.45mm filter is performed either in the lab or in the field. Using the Skalarautoanalyzer system, an inline reaction, of the ortho-phosphate ions with an acidicsolution containing molybdate and antimony ions, forms phosphomolybdic acid.Reduction of the phosphomolybdic acid by ascorbic acid forms an intensely bluecomplex. Measurement at 880 nm determines the concentration of reactedphosphorus.

WHP04: Hydrolysable PhosphorusFor the determination of dissolved hydrolysable phosphorus, filtration of the samplethrough a 0.45mm filter is performed either in the lab or in the field. Concentrations ofpolyphosphate and some organic phosphorus compounds are determined byconversion to ortho-phosphorus using acid hydrolysis. The Skalar autoanalyzer systemis used for inline hydrolysis using sulfuric acid added to the sample stream and heatedto 97°C. Colorimetric determination follows method WP04 , so WHPO4 includesreactive phosphorus.

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WTOTP: Total Phosphorus (UV Digestible)For the determination of dissolved total phosphorus, filtration of the sample through a0.45mm filter is performed either in the lab or in the field. Following hydrolysis(WHPO4 ), the sample undergoes further digestion with peroxodisulfate under UVradiation. Colorimetric determination follows method WPO4 .

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LABORATORY 7: Chlorophyll A

Trichromatic Method for Chlorophyll Analysis: (Strickland and Parsons,1968)

There are several methods of obtaining an estimate of primary productivity. Thefour most significant methods are counts, dry weights, turbidity and chlorophyll analysis.Each of these methods has advantages and disadvantages. Counts are generallyconsidered most satisfactory since only with this method are the species of algaeactually differentiated. However, the method is very slow and a great deal of theaccuracy depends on the operator. Dry weights and turbidity are both very fastmethods but it is difficult to distinguish the algae from the debris. Chlorophyll analysishas the advantage of being both fast and specific. Since it measures a component ofthe living cytoplasm, it gives an idea of the potential for growth of the algae.Unfortunately, it is difficult to correlate chlorophyll determinations with dry weightmeasurements or aerial standard unit counts or other assessments of productivity.

Spectrophotometric Determination of Chlorophyll a, b, c, and PlantCarotenoids:

The plant pigments of algae consist of the chlorophylls and carotenoids(carotenes and xanthophylls). The three major chlorophylls, a, b, and c absorb lightmaximally at specific wavelengths when dissolved in organic solvents. From theseabsorption characteristics, an estimate can be made of the concentrations of thepigments. Chlorophyll a is by far the most dominant chlorophyllous pigment and occursin greatest abundance. Therefore, chlorophyll a alone can be used to estimate algalbiomass. The spectrophotometric estimate of chlorophyll c concentration in thetrichromatic method below is only approximate. Using a more elaborate extractionmethod yields higher precision measurements of chlorophyll c (see Strickland andParsons, 1968); Chlorophyll concentrations are expressed in µg/L (or mg/m 3).Estimates of plant carotenoid pigments are reported collectively in pigment units thatapproximate mg/m 3 (Wetzel and Likens 1979).

A little more information:

Knowing the mass of chlorophyll a is very close to knowing primary production(Cole, 1975). Ryther and Yentsch (1957) experimentally demonstrated that a relativelyconstant relationship exists between chlorophyll and photosynthesis at any given lightintensity. Chlorophyll a is the master pigment in blue-green and eukaryotephotosynthesis. In a living cell, chlorophyll a absorbs light in two peaks - one between670 and 680 nm and the other at 435 nm. The longer wavelengths (670 – 680 nm) areabundantly present in shallow water while the shorter wavelengths penetrate deeper,thus allowing photosynthesis to occur at many levels (Cole 1975). Since plants containassortments of accessory pigments. Other light waves that travel vertically in the lakealso play a part in photosynthesis. Accessory pigment molecules absorb energy quantafrom light waves, become ‘exerted' and energy-rich, and pass their excitation energy on

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sequentially (Cole 1975). The energy eventually reaches chlorophyll a, the final energyrecipient found in all photosynthesizing plants. Chlorophyll b, c, and d have absorptionpeaks near but different from chlorophyll a. The result of all the organic pigments is thatenergy from light waves (range 400 nm to at least 700 nm) can be used in primaryproduction by higher plants (Cole 1975).

APPARATUS:25 mL graduated cylinder10 mL graduated cylinderPlastic filtration apparatus w/hosePlastic filter membranesHomogenizer unit with homogenizer tubes (Pyrex)Pipettes10 mL graduated centrifuge tubes

CentrifugeSpectrophotometerQuartz spectrophotometric cuvettes

MATERIALS AND REAGENTS:Algae culture (incubated for approx. 1 week)90% Acetone

METHOD:

1) Set up the filtration unit and using the fine tweezers provided, carefully place onecircle of the plastic membrane on the filter base. DO NOT touch the plasticmembrane with your fingers; it will disintegrate.

2) Connect the filtration unit to the vacuum and open the vacuum very slowly andslightly.

3) Swirl the sample culture vigorously and transfer 25 mL to a graduated cylinder.

4) Pour the 25 mL sample into the filtration unit.

5) Rinse the graduated cylinder and the unit with 1 - 2 mL of distilled water andallow the filter to dry (about one minute).

6) Using the tweezers, remove the plastic membrane from the filter base and placethe membrane in a homogenizer tube. Place the membrane as close to thebottom of the tube as possible.

7) Add 8 ml of 90% acetone to the homogenizer tube and macerate the membranecompletely using the homogenizer. In the process, the algal cells are dissociated

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and the pigments extracted. When maceration is complete, transfer the contentsto a 15 mL graduated centrifuge tube.

8) Rinse the homogenizer tube with 4 mL of 90% acetone and transfer the contentsto the same centrifuge tube.

9) Centrifuge the tube at approx. 2500 rpm (speed setting 1-2) until clear, about 20minutes.

10) Using a pipette, transfer about 5 mL of the supernatant to a quartzspectrophotometer cuvette and read the absorbance at 663, 645 and 630 nm onthe spectrophotometer in the instrumentation laboratory.

11) Record your observations and calculate the Chlorophyll a concentration in termsof mg CH a /L using the following equations:

Acorr = An - A750 (turbidity correction)

Correct for turbidity for each absorbance value first and then put the correctedvalues into the following equation:

A = (11.64 A 663 - 2.16 A 645 + 0.1 A 630 ) x 12.0 mL /0.025 L

For your report calculate the mg CH a /L for each group’s results and explain anydifferences observed. In addition, give a brief discussion of the advantagesand disadvantages of this technique

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LABORATORY 8: DETERMINATION OF HEAT BUDGET FORMIRROR LAKE

One of the most important and interesting characteristics of a lake is the thermalstructure. The heat content of a body of water is of vital importance in limnology.Directly related to the temperature of the aquatic environment are the metabolism,physiology and behaviour of aquatic organisms. Extreme temperatures restrict thegrowth and distribution of plants, animals and microbes (Wetzel and Likens 1979).Large volumes of water change temperature relatively slowly because of the highspecific heat of water. Therefore, large lakes tend to moderate local climates andprovide longer growing seasons for aquatic life, and serve as integrated recorders ofrecent climatic phenomena. These are some of the reasons that the thermal structureand heat content of a body of water must be known, with some degree of accuracy, inlimnological studies.

Determining a heat budget for 'Mirror Lake' requires a calculation of the heatcontent of the entire lake at the two specified time intervals.

Equation for the Heat Content of Lake Water (in calories):

The total heat content (storage) of the water on any sampling date may be determinedfrom:

zm θw = Σ tz Az h z

zo

θw = heat content of the lake water in calorieszo = surface of the lakezm = maximum depth of the laketz = average temperature in °C of a unit layer of water of thickness h z (in cm, with the midpoint at depth z)Az = the area at depth z in cm 2

(The heat content is usually expressed on a unit area basis, θw /Ao in cal/cm 2 where A o = surfacearea in cm 2)

Since the specific heat of water is one cal g -1°C -1 and since one cm 3 of water has amass of about one g, it is convenient to use a constant area of one cm 2 in calculations

of heat content for water, i.e. Az = 1 cm2

. However, because most natural lakes do nothave basins with perpendicular walls, it is necessary to correct for heat content thatvaries with depth. This may be done by using a ratio of the area at depth z to the areaof the surface of the lake. The ratio is usually obtained from a hypsographic orhypsometric curve.

Use the Sample data sheet below to calculate the heat content of Minor Lake for thetwo dates. Answer the questions below:

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Sample Data Sheet for the Heat Content Calculation:

* Total Column #6 to obtain the caloric (heat) content of the entire lake in cal/cm 2

Calculate the total heat content of Mirror Lake for the 17 th of May 1970 and 24 th of June 1970.What was the heat budget for Mirror Lake ( i.e. the difference in heat content over some timeinterval) over this interval? Did Mirror Lake gain or lose heat over this period?

4 5Reference Thickness of Weighting Average Caloric WeightedDepth (m) Layer (cm) Factor from Temperature (heat) Caloric (heat)

Hypsographic of Layer ( °C) Content Content/LayerCurve “Must be (cal/cm 2 of(% expressed calculated lake surface)as decimal T 1+T2/2 (Temp. x [(3) x (5)]fraction) (Temp. at Thickness)"For Mirror each end of [(2) x (4 )]Lake thecalculationsuse Table 4-2area %”

61 32

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N

E

S

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LABORATORY 9: WATER QUALITY ASSESSMENT

One of the many effects that organic wastes have on the aquatic environment isto influence the type of algae that will grow there. A survey made by Palmer (1969)revealed that more than 1000 algal taxa have been reported as pollution-tolerant forms.

The BOD (Biological Oxygen Demand) of the water measures organic “enrichment”, orpollution. The BOD describes how much oxygen is required for the biological oxidation(biodegradation) of the organic substances in the water. A high BOD indicates that therate of oxygen removal by bacteria and protozoa is high. Algal species tolerant oforganic pollution are significant in the recovery of a stream or lake because they addoxygen to the water during photosynthesis and incorporate organic and inorganicnutrients into their cells, thus removing pollutants from the water.

Algae is often used as a biological indicator species in determining the type anddegree of pollution m a body of water Three concentrated water samples will beanalyzed for their pollution levels as indicated by these species. To create the water

samples, 1 L of lake water was filtered for each 100 mL of concentrate (the concentrateallows the researcher to observe a large number of algae in a small sample). Theobjective is to count each of the algae in a known volume of sample, and then applyPalmer's (1969) pollution index to determine the level of pollution of the samples.Thirteen different types of algae have been identified in the samples provided. Youshould be familiar with all of them before leaving the lab today (Oil immersion will not benecessary for our purposes).

You will need:

One microscopeThree microscope slidesThree cover slips (25 mm x 25 mm)A keen eyeLots of enthusiasm!

You may work in groups of two, but no larger.

Follow this procedure for each sample:

1) Write your name and the name of your partner where indicated.

2) Indicate the sample number. You can get this from the bottle.

3) Examine the sample bottle. Pipette 0.1 mL of solution onto a slide and cover with thecover slip (Place the pipette as close to the bottom as possible). Try to avoid the largegreen masses of algae, as you will have a difficult time identifying individual species inthese clumps. Air bubbles are not allowed either.

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3) Determine the Pollution Index Value for each water sample. Palmer (19139)identified a list of organic pollution tolerant algae, to which he assigned PollutionIndex Values (Table 1). The Index can only be applied to those genera m agiven sample that have a concentration of > 50 individuals per mL of lake watersample (not the concentrate) Add up the Pollution Index Values for each of the

three samples. Record your results on Chart 2.A PI value of > 20 indicates high organic pollution. Midrange values (15-19)indicate that pollution is moderate. Low scores (

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Testing of Environmental Samples Using the LC50 QuantalTest

Toxicology studies, routinely carried out in the field of Limnology, are becomingincreasingly important for environmental protection. This lab exercise will introduce youto the LC50 test, a quantal test to determine the Lethal Concentration causing death in50 % of the test subjects, using neonate Daphnia spp. (

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Quantal Tests:

The example of a concentration-effect curve (or concentration-response curve) inFigure 1 represents results from an aquatic toxicology test. Figure l relates the intensityof the biological effect to the concentration in the test medium, and represents a lethality

test} this is a quantal test (all-or-none) in which each organism either shows the effect,or does not. The tested concentrations should produce several partial effects near themiddle of the percent scale, since such values have the least variability and the mostvalue or "weight" in estimating the nearby endpoint for 50% effect. There should alsobe a pronounced gradient of effect, ranging from little or no effect at a low concentrationto complete or nearly complete effect at a high concentration. Those very low and veryhigh percent effects have reduced weight in fitting the line, but they help to anchor it andestablish the slope, which is important in calculating confidence limits.

Figure 1 Results of a LC50 Test Plotted on a Logarithmic-probability Scale. A straight linehas been fitted to the data. The LC50 has been estimated graphically as about 5.5 mg/Lby the line drawn across from 50% effect, then vertically down from the intersection withthe fitted line. Probit analysis on computer confirmed the LC50 as 5.6% and estimatedthe 95% confidence limits shown by the horizontal bar at 50% effect.

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The median effect has the greatest precision as an endpoint for quantal tests.Familiar examples are the median lethal concentrations (LC50) and the medianeffective concentration (EC50). Half of the test organisms would have shown the effectat concentrations lower than the endpoint (C). The other half of the organisms wouldonly show effects at higher concentrations. The LC50's and EC50's always have a test

duration associated with them (e.g. 96-h LC50). For an EC50, the particular effectbeing tabulated must also be stated.

In quantal experiments, the endpoint is estimated by fitting a line to theconcentration-effect data. The LC50 could be read directly from an eye-fitted line(Figure 1). More formally, the estimation is made by probit analysis or by otherspecialized line-fitting procedures which use all of the data generated by the test.Accordingly, a line such as the one shown in Figure 1 is often referred to as a probitline. The mathematical model of the concentration-effect relationship also describes theassociated error term and estimates the precision of the endpoint, customarilyexpressed as the 95% confidence limits of the LC50 (or EC50).

Exposure time is an important component of all environmental toxicology tests,and quantal tests are no exception. If the results shown in Figure 1 were taken to bethose of all acute lethality tests, a series of tests with different exposure times would beexpected to result in a series of different probit lines. Short exposures would requirehigher concentrations to manifest mortalities from 0 - 100%. Long exposures would beexpected to require lower concentrations. However, in acute testing, there is often anexposure time at which the maximum acute effect has been achieved. In other words,prolonging the exposure would not increase the magnitude of the acute effect.

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Toxicology in Aquatic Environments

1. Define Toxicity; distinguish between acute toxicity and chronic toxicity.

2. Describe how non-polar substances differ from polar substances in theirbehaviour in the environment and organisms.

3. What is an LC50?

4. Explain the difference between a high K ow and a low K ow.

5. Define Bioaccumulation, Biomagnification, and Bioconcentration.

6. What are three organisms commonly used in aquatic toxicity testing?

7. Why is a plot of log-transformed dose vs. probit-transformed response usedrather than dose vs. response in determining an LC50 or LD50.

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Sections:

Title

The title of a report/article is the first part of your report that will be read – andpossibly the only part read for journal articles. Therefore, the title must serve twopurposes ; a title informs the reader about the subject of the study, anddistinguishes your work from similar studies in the literature.

Titles are not necessarily complete sentences; titles are pared down to containonly the words essential to conveying the important details . There should be aclear relationship between all of the details mentioned in the title, and althoughthe title should distinguish your work from others, trying to cram too much detailinto a title will make unreadable. Browsing scientific journals – especially thosein your field – will give you a good feel for a properly constructed title.

Abstract:

The abstract is a summary of the report, and should give the reader the followinginformation – usually presented in this order:

The subject of the report

The main objectives

Brief description of the methods

Summary of the most important results

The major conclusions and significance

Abstracts are usually under 300 words in length and must be concise ; do notinclude references or citations in an abstract and only include information foundin the body of the report. Readers of scientific journals will typically only read thetitle and the abstract – if the title was appealing enough –, so a good abstract isimportant for getting your article read.

Introduction

This section sets the stage for the rest of the report. The introduction informs thereader about the scope, objectives, limitations, structure and importance ofthe study/report. Any jargon or specialized terms should be introduced in the

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introduction – acronyms used should be written out in full followed by theacronym in parentheses – as well as any concepts discussed later in the report. Do not fill the Introduction with definitions from the glossary.

There are usually a large number of literature citations in the introduction –

usually half of the citations are in this section. Students usually cite fewerreferences than is appropriate, but it is a writing skill that will improve withexperience – more discussed in the References section.

The length of the introduction will vary with the material introduced; theintroduction should be concise, but should be long enough to fully prepare thereader for the reported study.

Results and Discussion

The results and discussion of a report/article are where you get to present yourfindings. The previous sections of the report prepare the reader for what youshow them in this section; the readers are now familiar with the background andsubject of the study, the techniques used, and why the study was performed. Allthat remains for this section of the report is for you to point out the importantfinding s and discuss their significance within the scope of the study.

There are two common styles for this section; the results and discussion can becombined into one large section or they can be written as two separate sections. If writing in the combined style, the results can be interpreted as theyare presented – if interpretation is not dependent on a latter result. Otherwise,

the findings are presented in the results section, but with no interpretation, which is done in the discussion section.

Regardless of the chosen style, the results must include text . Readers mustbe guided through the results section and directed to the important details in theillustrations - simply telling the reader to "See Table (Figure)..." is not sufficient.Readers need to be told : what to look at, what it means, why it is important, andthe implications of the findings – when interpretation is done.

Interpretation includes discussing the significance and implications of the resultswithin the scope of the report, and relating your findings to those of other studies

in the literature. Discrepancy between your results and those of other studiesneed to be explained - . Do not make generalized statements that are not basedon your data or known fact, however.

There is some choice for how the findings are presented; results can be part ofthe text , displayed in tabular form, or compiled into a figure/graph . Thepresentation choice is determined by what you are trying to say about the dataand how appropriately the text/table/figure gets your point across. All

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Do not use footnotes; place your references in the literature cited section. Inscientific papers, full quotes are not used; material is paraphrased and areference is cited at the appropriate place. Citations are usually placed at theend of the sentence, but if several bits of information are used from the samesource, then the citation can be placed at the end of the paragraph. It is also

appropriate to include the citation as part of the sentence, e.g. it was noted bySmith et al. (2003) that the sky is blue.

Note: For a book, the total number of pages is listed, but for a journal article thearticle page numbers are listed.

Why Use References?

A common problem with student papers is plagiarism, i.e. presenting the ideasand findings of others as if they were your own without credit/references. Usuallythis is unintended plagiarism; inexperienced students may not recognize the

material needs a reference. Unless something is common knowledge (e.g.birds can fly) or it is your own information – unlikely unless you have had aresearch career prior to the course –, you must give credit to the authors .

Plagiarism also applies to copying another student’s paper and submitting it asyour own. Additionally, you cannot resubmit a paper you wrote for another courseor in another year.

Plagiarism is a serious academic offence . Plagiarism is defined in UniversityRegulation IX Academic Dishonesty as:

1. Plagiarism of ideas as where an idea of an author or speaker is incorporatedinto the body of an assignment as though it were the writer's idea, i.e. no credit isgiven the person through referencing or footnoting or endnoting.

2. Plagiarism of words occurs when phrases, sentences, tables or illustrations ofan author or speaker are incorporated into the body of a writer's own, i.e. noquotations or indentations (depending on the format followed) are present butreferencing or footnoting or endnoting is given.

3. Plagiarism of ideas and words as where words and an idea(s) of an author orspeaker are incorporated into the body of a written assignment as though they

were the writer's own words and ideas,i.e.

no quotations or indentations(depending on format followed) are present and no referencing or footnoting orendnoting is given.

From University Regulations – IX Academic Dishonestyhttp://calendar.lakeheadu.ca/current/contents/regulations/univregsIXacdishon.html

Additionally, plagiarism is considered misconduct under Code of StudentConduct and Disciplinary Procedures (http://policies.lakeheadu.ca/policy.php?pid=60 ).

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The minimum penalty for a student found guilty of plagiarism is a mark of zero for the work concerned. Serious or repeated plagiarism can result inexpulsion from the University and a mark of zero for the course – a mark that isoften used by Universities to note academic dishonesty.

Some Tips

1. Please, please, pretty please PROOF-READ!!

2. Read your work out loud, does it make sense?

3. Scientific names are in Latin/Greek, you must Italicize or Underline!!!

4. You can refer to an organism by Genus first letter, period, species name, e.g.

G. species , after you have written it out in full once .

5. Alternatively you could refer to an organism by its common name as long as

you associate it with the proper scientific name first. E.g. Rainbow trout

(Oncorhynchus mykiss ).

6. G enus is Capitalized, s pecies is n ot.

7. Check for correct use of a ffect/ e ffect.

8. Also check for the ir /the re /the y’re .

9. And for It ’s (It is, It has)/Its.

10. And for lose/lo o se (trust me; it happens more than you think).11. Don’t use contractions.

12. Proof-Read!

MarkingThe marking scheme for reports will be as follows:

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Appendix 2

Worksheets for Laboratory 9

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ALGAL SPECIES FOUND NOT FOUND

CLOSTERIUM

SYNEDRA

MICRASTERIAS

CHLORELLA

STAUSTRUM

PHACUS

EUGLENA

CHLAMYDOMONAS

PEDIASTRUM

SCENEDESMUS

PANDORINA

OSCILLATORIA

BULBOCHAETE

STIGEOCLONIUM

NAME:___________________________________________

NAME OF PARTNER:_______________________________

SAMPLE NUMBER:_________________________________

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NUMBERALGAL SPECIES

STRIP 1 STRIP 2 STRIP 3 STRIP 4

CLOSTERIUM

SYNEDRA

MICRASTERIAS

CHLORELLA

STAUSTRUM

PHACUS

EUGLENA

CHLAMYDOMONAS

PEDIASTRUM

SCENEDESMUS

PANDORINA

OSCILLATORIA

BULBOCHAETE

STIGEOCLONIUM

NAME:___________________________________________

NAME OF PARTNER:_______________________________

SAMPLE NUMBER:_________________________________

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ALGAL SPECIES FOUND NOT FOUND

CLOSTERIUM

SYNEDRA

MICRASTERIAS

CHLORELLA

STAUSTRUM

PHACUS

EUGLENA

CHLAMYDOMONAS

PEDIASTRUM

SCENEDESMUS

PANDORINA

OSCILLATORIA

BULBOCHAETE

STIGEOCLONIUM

NAME:___________________________________________

NAME OF PARTNER:_______________________________

SAMPLE NUMBER:_________________________________

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NUMBERALGAL SPECIES

STRIP 1 STRIP 2 STRIP 3 STRIP 4

CLOSTERIUM

SYNEDRA

MICRASTERIAS

CHLORELLA

STAUSTRUM

PHACUS

EUGLENA

CHLAMYDOMONAS

PEDIASTRUM

SCENEDESMUS

PANDORINA

OSCILLATORIA

BULBOCHAETE

STIGEOCLONIUM

NAME:___________________________________________

NAME OF PARTNER:_______________________________

SAMPLE NUMBER:_________________________________

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ALGAL SPECIES FOUND NOT FOUND

CLOSTERIUM

SYNEDRA

MICRASTERIAS

CHLORELLA

STAUSTRUM

PHACUS

EUGLENA

CHLAMYDOMONAS

PEDIASTRUM

SCENEDESMUS

PANDORINA

OSCILLATORIA

BULBOCHAETE

STIGEOCLONIUM

NAME:___________________________________________

NAME OF PARTNER:_______________________________

SAMPLE NUMBER:_________________________________

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NUMBERALGAL SPECIES

STRIP 1 STRIP 2 STRIP 3 STRIP 4

CLOSTERIUM

SYNEDRA

MICRASTE