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 of
that story of Narcissus, who because he could not grasp the tormenting, mild image he saw in the
fountain, plunged into it and was drowned. But that same image, we ourselves see in all rivers and
oceans. 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 3rd ed., R. W. Wetzel. 2001. Academic Press. San Diego,
CA.
LECTURE TOPIC
1. Importance of Limnology
2. Lake Formation
3. Light
4. Heat Budgets of Lakes
5. Water Movement
6. Oxygen
7. Carbonate Cycle
8. Chemical Constituents of Lakes
9. Phytoplankton and Zooplankton Ecology
10. Productivity of Aquatic Systems

Chapter 1
Chapter 3
Chapter 5
Chapter 6
Chapter 7
Chapter 9
Chapter 11
Chapter 12,13,14
Chapter 15
Chapter 24, 25

LABS
Please 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)

BIOLOGY 4710- Limnology Laboratory Schedule Winter 2006

Lab:

Description:

Date:

Field Trip Preparation

Jan .9

Map production (assignment)

Jan. 16

Field I rip (Kingfisher Lake)

Jan. 23

Water Quality Analysis Alkalinity

Jan. 30

5&6

Total Kjeldahl Nitrogen and Phosphorus

(Combined Lab - only 1 report req'd)

Feb. 6

Chlorophyll a

Feb. 13

Lake Capacity/Heat Budgets

Feb. 27

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
Abstract
2
Introduction
4
Results
5
Discussion/Conclusions
4
Maps
3
Presentation
2
Total

20

Labs
1
2
3
4

10

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 was
carried out over and a summary of the major findings, including the morphological data
gained from the field trip preparation and water quality results obtained from the field
trip.
The report should also include all of the information requested in the laboratory
manual, but not details of the use of equipment. Equipment may be discussed if more
than 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 more
than 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 clear
titles, 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 limnological
studies of lakes and the design of the survey, i.e. which tests were carried out, what
data was collected (where, when, and why). This section should also include a
description of the lake, including the location and access, and should be no more than
two pages plus a map.
Results - This section should be the major section in the report and contain maps or
diagrams with some discussion. This section should be divided into Morphological
Attributes, 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 suffering
noticeable signs of deterioration or are there any problems with water quality. This
section should not need to be more than two pages.
References This should follow the guidelines from the journal Limnology &
Oceanography

INTRODUCTION
The collection of accurate limnological data from water bodies is necessary for
an understanding of the status of wetlands. Lake classification systems based on
trophic 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; measurement
of physical characteristics such as temperature profiles, colour and turbidity; and
chemical parameters such as pH, dissolved oxygen, alkalinity, concentrations of
inorganic and organic compounds and the composition and biomass of micro flora and
fauna.
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 and
water quality parameters. This will also be an opportunity to practise water and
sediment sample collection techniques.

LABORATORY 1: Field Trip Preparation

Morphometry is defined as the methods of measuring and analyzing the physical
dimensions of a lake or stream (Coke 1994). In order to carry out limnological analyses
of a lake, a detailed knowledge of the morphometry should be established, with
particular emphasis on the volume characteristics (Wetzel and Likens 1979). Critical
components of detailed analyses of biological, chemical and physical properties of fresh
include depth analyses (including area measurements of sediments and water strata at
various depths), volumes of strata and shoreline characteristics (Wetzel and Likens
1979). For example, morphometric parameters are required to evaluate erosion,
nutrient loading rates, chemical mass, heat content and thermal stability, biological
productivity, effectiveness of growth and other structural and functional components of
the ecosystem (Wetzel and Likens 1979). Detailed knowledge of morphometry and flow
characteristics of freshwater bodies is relied upon in management techniques, such as
the loading capacity for effluents and the selective removal of undesirable components
of the biota (Wetzel and Likens 1979).
Detailed hydrographic maps of lakes and streams are often unavailable to the
limnologist. Those available from governments or other sources must be checked for
accuracy, as the morphometry of a body of fresh water changes with time. It is
therefore of great importance for the student to have a general understanding of the
construction of bathymetric (contour) maps and the computation of morphometry
parameters (Wetzel and Likens 1979)
Before any field trip to survey the limnological parameters of a water body, a number of
preparatory 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 are
to be measured from collected samples.
7) Choose and familiarize yourself with the equipment needed to collect the data
and 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 body
may require the use of the Gazetteer, Topographical maps, National
Watershed Code Maps and Forest Resource Inventory Maps (FRI).

The Gazetteer is the official source of information in Canada giving the

name of the lake that corresponds to the geographical coordinates found
from a Topographical map The latest edition is 1974, with subsequent
supplements.
Write down the name of the lake that you will be surveying.
The 1:50,000 Topographical Map is the most common and available
source of information for identifying and distinguishing water bodies. Be
sure to check the scale of the map you are working from!
Write down the geographical position, general location and extent of
the lake.
1.1.2.

Elevation: The elevation is measured as the height above sea level of ft

water 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 elevations
of connecting lakes. Elevation in feet has to be converted to meters
Give the elevation of the lake.

1.1.3.

Watershed Code: The Watershed Code Maps, number watersheds and

drainage systems m Canada. In Ontario, there are three main drainage
systems: The St. Lawrence (2), The Hudson Bay (4) and Lake Winnipeg
(5). Each system has been divided into Principal Watersheds and
Watershed 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 reaching
a 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

*NOTE: You may combine 5) with 1), 2), and 3) H the final map produced is a clear and
neat presentation. You should have a separate rough draft contour map.
1.2.1.

Tracing the outline Map: The most accurate production of maps is by

the digital scanning of aerial photographs or through direct access of
digital satellite images that can be geocoded by means of a Global
Positioning System (GPS). For our purposes, the accuracy of tracing an
outline from a Forest Resource Inventory (FRI) map, followed by enlarging
from an overhead protector, is adequate. The FRI maps are traced from
aerial photographs at four inches to one mile or 1:16,000 and are thus
more accurate than the Topography maps, which are 1:50,000.
Place a piece of tracing paper over the FRI map and carefully trace
an image of the study lake.
Other methods of enlarging the traced image include the use of a
pantograph (hinged parallelogram) however, for our purposes; the use of
an overhead projector is adequate Place the traced image on an overhead
projector with a scab and north pointer. Note the enlargement ratio from
the projected scale and draw the projected image on to map paper.*
*NOTE: This step may not be required if the map being used is at a
1:20000 scale.

Lake Outline Map

Gazetteer name

Jones Lake

Local name

Jones Lake

MNR District

Chatham

Township

Elgin

Lat. and Long.

4217 - 8119

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 perimeter
of 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. The

indicator needle is set at zero by rotating the tracing wheel.
Measure the shoreline by tracing the outline of the figure and then
read the result from the scale. Care must be taken so that the
indicator needle travels In a positive direction. Also on a large lake
the needle may travel more than once around the circumference of
the dial. Therefore, the operator should keep close watch as to the
number of revolutions.
Use a start/finish line on the map to eliminate most of these
problems.

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 area
can be estimated from a dot grid overlay but for our purposes, we will use
the 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 km2.

Using the Digital Planimeter (Calculation of Lake Area)

1) Choose starting point on the map - centre the tracer lens on this
point. Press ON.
2) Press UNIT key until km2 is highlighted.
3) Enter the 1 : N scale value of your map by typing in N. i.e. If your
scale 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 your
starting position. Once you are back at your starting position press
HOLD.
6) Move the tracer lens to the start position on any islands in the lake
and 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 the
areas. Remember to press END after each tracing.
12) Calculate the average area.
NOTE: For later calculations you will need to remember the following:
1 km2 = 100 ha

LABORATORY 2: Field Activities

Three exercises have to be completed on the field trip. The data that is collected will be
used in a report to be handed in on Friday November 9. This report will include
information 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
Head Lake

deep holes. To prepare the Contour Map, the various

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

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

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 by
reading the appropriate Instruction Manuals. Only some common features are
expressed 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

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.

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 Procedures
Echo 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

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.

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.

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 profile
7) 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 O2, pH and conductivity will be done as one large group.

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 the
fast 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.

2.2.1 Time of Day

Water chemistry measurements in lakes are performed during the solar mid-day
hours (i.e. between 1130 hours and 1400 hours).
2.2.2 Water Transparency
Transparency describes the extent of light penetration into water. As light plays
an important part in many biological processes, this measurement must be performed
accurately. Transparency is always measured at the time of every chemistry test. If the
chemistry tests are done before noon, then transparency tests are performed upon
completion of the chemistry tests. If the chemistry tests are done in the afternoon,
transparency is done first, to get the reading as close to high noon as possible.
2.2.3 Secchi Disc

Transparency in lakes is measured with a weighted metal disc, twenty

centimetres in diameter, with alternating white and black quadrants. This instrument is
known as a secchi disc. A line, graduated in metres and centimetres is attached to the
disc.
To obtain a Secchi disc reading:
1)

Lower the disc into the water on the shady side of the boat and note the
depth at which the disc disappears from view in descent.

2)

Raise the disc and note the depth at which the disc reappears in ascent.

3)

Calculate and record the arithmetic mean of these two readings. This is
the secchi disc reading.

4)

Record water surface conditions cloud cover and time as listed below.

Note: Do Not Wear Sunglasses when taking secchi disc readings.

2.2.4 Water Surface Conditions

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 (instructions
for completion of calculations are given in the following pages).
1) Areas of the surface and each contour interval at depth z (contour areas for each
contour)
2) Lake Volume
3) Mean Depth
4) Shoreline Development Factor
5) Maximum Depth

3.1

PRODUCING THE CONTOUR MAP

The contour map is prepared by transcribing the depths from the echo sounding tapes
Short Tips

Setting Knob

Adjusting
Knob

Fulcrum

Proportional Dividers

Long Tips

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

(described further on) will be used for the remaining transects. It will not be as accurate
but 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 to
measure ratios or proportions.
The tips on each end of the arms are of different lengths. One end has long tips and
the other end has short tips (See diagram.) The fulcrum also can be adjusted and
moved 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 the
length of the longer line and at the same time the ends with the shorter tips extend to
the length of the shorter line. (See diagram).
NOTE: Adjusting the dividers to the correct position can be tricky.
See the following instructions.
Adjusting the Dividers
To 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 can
then 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-consuming
measurements and calculations and therefore is not recommended.
2) Set the dividers by trial and error. This method is quick and involves no
measurements or calculations
Trial and Error Method

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.

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.
Fulcrum

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 with
experience.
In addition, if the same general boat speed has been maintained for each sounding
transect, then very little adjustment of the proportional dividers will be necessary for
each transect, as the ratio will be consistent.
When lines are too long for the dividers, divide the lines into equal sections and do
separately.

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.

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) = Ratio
Tape 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

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.

INITIAL CONTOUR MAP

The Final Contour Map

Once the initial contour map is completed, it is traced to produce the final map which will
not 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 = 104 m2
1 km2 = 100 ha
3.2.2. Volume Calculation

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(104m3)
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.

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. =

2A
P = shoreline perimeter (km)
A = area (km2)
= 3.14
3.2.5. Maximum Depth
Maximum depth in metres is the greatest depth recorded for the whole
lake.

LABORATORY 4: WATER QUALITY ANALYSIS

Chemical Analysis
Precipitation falling upon the surface of the earth, as either rain or snow, contains
a variety of dissolved gases. Additionally, incorporation of aerosols or dust particles
occurs along the way. Therefore, by the time the water reaches the earth's surface, it is
no longer pure. As it flows over, or penetrates into plants and soil, water dissolves more
gases. Notably, carbon dioxide and various mineral substances with which it has come
into contact. CO2 dissolves in water, forming carbonic acid and lowering the pH:
H2O + CO2 H2CO3
Most minerals are only slightly soluble in water. For example, limestone (calcium
carbonate, CaCO3) is slightly soluble in pure water. However, limestone is more soluble
in water containing carbonic acid; the conversion of CaCO3 to calcium bicarbonate
allows more limestone to dissolve:
CaCO3 + H2CO3 Ca(HCO3)2
(Insoluble)
(Soluble)
The dissolved bicarbonate has a marked effect on the chemical properties of the

+
water. Ca(HCO3)2, for example, dissociates into Ca2 and HCO3 ions. The HCO3
+
ions react with H (which originated from the natural dissociation of H2O) to form
carbonic acid (H2CO3). The H2CO3, in turn, dissociates into soluble CO2, which is often
in equilibrium with CO2 from the air and H2O. Therefore, the bicarbonate changes the
pH 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 an
important factor in the treatment of the water for domestic and industrial use.
In addition to bicarbonates, carbonates, and hydroxides, other moderately
soluble minerals are silica, chlorides, sulfates, and nitrates of calcium, magnesium,
sodium and potassium. With increased emission of sulfur dioxide (SO2) into the

atmosphere from industrial activities, SO4 is becoming the dominant anion in

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

In natural waters, inorganic carbon as dissolved CO2 and HCO3 is the primary
carbon source for photosynthesis by algae and larger aquatic plants. In addition to

respiratory production of CO2 by most organisms, influxes of CO2 and HCO3 from
incoming water and from the atmosphere balance this utilization. The amounts of
bioavailable inorganic carbon are adequate in most natural fresh waters; only under
special conditions, such as soft waters and intensely productive situations, does
inorganic carbon become a limiting factor to photosynthesis.

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 the
ratios of one to another determine the observed pH and the efficiency of buffering of a
given 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 more
restrictive. Therefore, the capacity of natural waters to resist changes in pH is very
important to the maintenance of life.
Alkalinity of fresh waters refers to the measure of the capacity of water to
neutralize acids. The main contributors to alkalinity in water are bicarbonates,
carbonates, and hydroxides; and less frequently by borate, silicate, and phosphate.
Since CO2 is relatively abundant in both gaseous and dissolved form, and bicarbonates
and carbonates are common in primary minerals over wide areas of the earth,
carbonate anions usually dominate the buffering system of fresh waters. Direct
contributions to alkalinity by hydroxides are rare in nature, except in very nutrient-poor
waters.
The interrelationships between carbon dioxide and the other major components of
alkalinity are as follows:
Free CO2 in the air is often in equilibrium with dissolved CO2 in the water. This
equilibrium, together with the other equilibria taking place in the water, is
represented by the following equation:
+

CO2 (air) CO2 (dissolved) + H2O H2CO3 H + HC03

H+ + CO32

The concentration of CO2 in the atmosphere averages about 0.03 percent, but
varies with location. Photosynthesis and respiration by aquatic organisms substantially
influences the quantity of CO2 in water at any given time and place. After equilibrium is
established in the water, the resulting condition is exemplified by the equations:

CO2 + 2H2O HCO3 + H3O

HCO3 + H2O H2CO3 + OH

H2CO3 H2O + CO2

(1)
(2)

(3)

The hydroxyl ions (OH ) formed (equations 1 and 2) show why waters with high
+
carbonate content are alkaline. Acid (source of H ) must be added to bring the water to
the point of neutrality, i.e. where equal quantities of hydronium and hydroxyl ions are
present.
The equilibria shown by equations 1, 2, and 3 explain the buffering capacity of
alkaline waters. That is, the water tends to resist change in pH as long as these

equilibria are in existence. Addition of H (Acid) reacts with the OH formed in equation

2; 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 supply
of carbonate and bicarbonate is exhausted. Similarly, addition of OH- initiates the
following reaction:

HCO3 + OH

CO3 + H2O

(4)

The terms alkalinity, total alkalinity, alkaline reserve, titratable base, or acidbinding capacity are frequently used to express the total quantity of base, in equilibrium
with 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 titration
with a standard solution of a strong acid, e.g., 0.02N H2SO4, to certain equivalence
points as given by indicator solutions. The indicator phenolphthalein is commonly used
for 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 to
measure the alkalinity contributed by bicarbonate (See figure below).
To facilitate calculations, all alkalinity sources are expressed as either
milliequivalents per Litre or milligrams calcium carbonate per Litre. The latter is termed
Total Alkalinity as calcium carbonate. From a practical standpoint, this method of
expressing alkalinity is satisfactory and is used extensively in limnology.

Relation between pH and the relative proportions of inorganic carbon

2
species of CO2, HCO3 , and CO3 in solution (From Wetzel, 1975)

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

The pH of a water body is a measure of the hydronium ions present in solution. In

lakes, 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 usually
measured directly in the field because it is readily altered by biologically activity and
temperature.
Conductivity
Conductivity is a measure of the electrical resistivity of a solution. Conductivity depends
on the presence and concentration of ions, particularly the simple inorganic ions Na+,
Ca2+, Mg2+, K+, CI-, HCO-, and sulfur based oxoanions (SOxy). Higher concentrations of
ions mean lower electrical resistivity and therefore, higher conductivity. Additionally,
conductivity can measure salinity, because salinity is the total concentration of ions in
solution (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 a
defined temperature usually 180 C , of a filtered sample of known volume. In
addition, conductivity can estimate TDS via multiplication by a conversion factor
usually 0.6.
Dissolved oxygen
The distribution of O2 in natural waters provides a measure of organic production and
decomposition, and is a basis for most methods of measuring primary productivity. The
Winkler titration method is routinely used for measuring oxygen concentration. It is a
simple oxidation-reduction reaction involving manganese, iodine and thiosulfate.
Alkalinity
Alkalinity is the common method for determining carbonate content and the buffering
capacity of water. Additionally, alkalinity is an index to the rock contents within a
drainage basin and the degree to which they are weathered. Limestone present in
drainage basins will increase the carbonate content through dissolution of CaCO3 and
therefore 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 bromcresol
green - methyl red).

In environmental literature, two kinds of alkalinity are usually reported: phenolphthalein

alkalinity and total alkalinity. Phenolphthalein alkalinity (P) measures the buffering

action of bases as strong as, or stronger than, the carbonate ion (CO32 ).
Phenolphthalein alkalinity exists when the pH is greater than 8.3. When
phenolphthalein 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. This
represents all of the hydroxide alkalinity, 1/2 of the carbonate alkalinity, and 1/3 of the
phosphate and any other alkali producing material present in the sample above a pH of
8.3. There is usually no hydroxide alkalinity in water of pH less than 9.2. If the sample
water is initially below pH 8.3, the phenolphthalein alkalinity is zero.
Sample reactions that occur during the titration include:

OH + H (from sulfuric acid) H2O

CO32 + H

HCO3

Adding bromcresol green-methyl red indicator to the water sample will turn it a bluegreen color. Bromcresol green-methyl red measures the buffering action of bases as

strong as, or stronger than, the bicarbonate ion (HCO3 ). Adding acid to the sample will
change the colour to pinkish-purple when a pH of 4.3 is reached. The following reaction
takes place during Bromcresol green-methyl red alkalinity determination:

HCO3 + H (from sulfuric acid) CO2 + H2O

Total (T) alkalinity is the sum of the phenolphthalein alkalinity and the bromcresol greenmethyl red alkalinity. This represents all of the hydroxide, all of the carbonate, and 2/3
of the phosphate and other alkali producing material present in the sample above a pH
of 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 mineral
acids.

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

APPARATUS:
50 mL burette stand and holder
250 mL Erlenmeyer flask funnel
Pasteur pipette

REAGENTS:
0.02N H2SO4
Phenolphthalein indicator
Bromcresol green-methyl red indicator

METHOD:
1)

Assemble the titration apparatus Rinse the burette with a few mL of 0.02N
Sulfuric acid and then fill the burette to the 50 mL mark. Use caution and
a funnel.

2)

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

3)

Add 4 to 5 drops of phenolphthalein indicator to the sample. If a pink

colour appears, add the sulfuric acid slowly until the pink colour
disappears upon swirling, Note the volume of acid used, this volume is
required to calculate the phenolphthalein alkalinity.

4)

Then add 3 to 4 drops of bromcresol green-methyl red indicator to the

same sample. Slowly continue to add the sulfuric acid to the appropriate
equivalence point (faint pink). The colour change will be green to clear to
faint pink. Remember to swirl the flask after each addition of acid and
when you are approaching the endpoint, add the acid drop-wise. Note the
total volume of acid used; this is required for the calculations of total
alkalinity.
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)

7)

Calculate the phenolphthalein and total alkalinity for the water samples
using the following equations:
Phenolphthalein alkalinity as mg/L CaCO3
Volume of acid used to first endpoint (mL) x (normality of acid x 50000)
Volume of sample (mL)
Total alkalinity as mg/L CaCO3

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

LABORATORY 5: TOTAL KJELDAHL NITROGEN

Compounds of nitrogen and phosphorus are major cellular components of
organisms. Since the availability of these elements may be less than biological
demand, then environmental sources can regulate or limit the production of organisms
in freshwater ecosystems.
Concentrations of nitrogen and phosphorus compounds are highly dynamic
because they may be utilized, stored, transformed and excreted rapidly and repeatedly
by various aquatic organisms. Measurements of these elements are complicated by the
chemical form in which they occur. Ionic concentrations are often very low, requiring
care in collection and analysis of water samples, to avoid contamination.

Total Kjeldahl Nitrogen

Total Kjeldahl Nitrogen (TKN) includes the organic nitrogen compounds plus the
ammonia fraction but does not include NO3 and NO2 present in a water sample. The
procedure has been developed by L.U.E.L. and is as follows:
WTKN: Total Kjeldahl Nitrogen
Determination by the difference of WTOTN and WNOX
WTKN = WTOTN - WNOX
WTOTN: Total Nitrogen (UV digestible)
Using the Skalar autoanalyzer system, the sample is mixed with a potassium
peroxodisulfate/sodium hydroxide solution and heated to 90C. The solution is then
mixed with a borax buffer and all nitrogen species are converted by UV radiation to
nitrate. Colorimetric determination follows WNOX. This method accounts for nitrogen in
the form of nitrate, nitrite and a variety of other forms.
WNOX: Nitrate and Nitrite
Using the Skalar autoanalyzer system, the nitrate is reduced to nitrite in a commercially
packed cadmium column treated with copper sulfate. All nitrite is then determined by
diazotizing with sulfanilamide and coupling with N-(1-naphthyl)-ethylenediamine
dihydrochloride to form a highly coloured azo dye (Griess reaction). The absorption is
measured 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 to
calculate the value for Kjeldahl Nitrogen for your report.
Discuss TKN to an appropriate extent in your report and be sure to include a discussion
on the reasons for variability in your results (if any) between the three depths sampled.

LABORATORY 6: TOTAL PHOSPHORUS

Phosphorus plays a major role in metabolism in the biosphere and is of significant
interest ecologically. There is a relatively rich supply of other major nutritional and
structural components of the biota (C, N, O, and S); however, phosphorus is the least
abundant of the components and consequently limits biological productivity. Unlike
nitrogen, phosphorus does not have a large immediate storage reservoir, the
atmosphere.
Phosphorus occurs predominantly as phosphates in both natural waters and
wastewaters. There are three classifications of phosphates: condensed phosphates,
ortho-phosphates and organic phosphates. Phosphates occur in solution, in adsorbed
to 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 supplies
during treatment
Larger quantities added when the water from
laundering or cleaning (major constituents of many
commercial cleaning compounds)
B) Ortho-phosphates

Applied agricultural or residential fertilizers,

transported to surface waters via storm runoff and
spring melt

C) Organic Phosphates

Formed primarily by biological processes, form a part

of sewage through body wastes and food residue
May also be formed from orthophosphates in
biological treatment processes or by receiving water
biota

The stimulated growth of photosynthetic micro- and macro-organisms to nuisance

quantities, in phosphate limited aquatic systems, stems from the discharge of raw or
treated 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

Samples collected from Kingfisher Lake have undergone a sulfuric acid digestion
procedure prior to the lab. This digestion oxidizes organic matter, to release
phosphorus as orthophosphate. The digested samples are ready for total phosphorus
analysis using a colorimetric determination of dissolved orthophosphate. The intensity
of colour, indicated by absorbance (ABS) values, is proportional to the amount of
phosphate present in a sample. A standard curve and regression equation is calculated
by performing a linear regression analysis, on the ABS values obtained for known
standards using a spectrophotometer. The regression equation determines unknown
concentrations of phosphorus in your sample as a function of the sample ABS values.

Determination of Phosphorus (Phosphorus in water by the Ascorbic Acid

Method)
To create the standard curve, four phosphorus standards are used.
Standard Values in mg/L
S0 = 0.0
S1 = 0.020
S2 = 0.040
S3 = 0.060C
NOTE: Safety glasses MUST be worn when working with acids (the Combined
Reagent includes acid).
Combined Reagent:
30 ml Ammonium Molybdate solution
100 mL 5N H2SO4
60 ml Ascorbic Acid Solution
10 ml Potassium Antimony Tartrate
To each of the Standard Solutions and digested samples, add 4.0 mL of Combined
Reagent 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, because
dissolved gas interferes with the absorbance measurement. After 10 minutes, but
within 2 hours, pipette the solution into a cuvette and read the absorbances.
Absorbance at 880 nm (A880) is measured on the CARY 50 Spectrophotometer with 10
mm cuvettes. This instrument is a single beam spectrophotometer that measures the
difference between a reference solution and a sample solution.
The CARY is zeroed using a 0.0 mg P/L reference solution of degassed DDW. Once
the CARY has been zeroed, the standards and samples can be measured.

Measure the Absorbance (A880) of each standard, from lowest to highest concentration.
Rinse the cuvette with degassed DDW between each sample and discard the rinsate in
the 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 Kimwipes only. Continue the analytical run with your samples.
Once you have obtained your A880 values for the standards and samples, perform a
linear regression analysis to calculate the concentration of phosphorus, as a function of
the absorbance units.
To create a standard curve, use the concentration of phosphorus as your X-axis
(independent variable) and the A880 as your Y-axis (dependent variable); plot the data
for the phosphorus standards. Once you have plotted your regression line, you can
determine the concentration of Total Phosphorus for your samples. By plotting your
data you can then estimate on your regression line what the concentrations should be
for 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 thermal
layers will give results that are more accurate. Include your regression line and
calculated Total Phosphorus results as an appendix to your report. The results from the
Skalar Autoanalyzer should be used for your report as these results will be the most
accurate. 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 through
a 0.45mm filter is performed either in the lab or in the field. Using the Skalar
autoanalyzer system, an inline reaction, of the ortho-phosphate ions with an acidic
solution containing molybdate and antimony ions, forms phosphomolybdic acid.
Reduction of the phosphomolybdic acid by ascorbic acid forms an intensely blue
complex. Measurement at 880 nm determines the concentration of reacted
phosphorus.
WHP04: Hydrolysable Phosphorus
For the determination of dissolved hydrolysable phosphorus, filtration of the sample
through a 0.45mm filter is performed either in the lab or in the field. Concentrations of
polyphosphate and some organic phosphorus compounds are determined by
conversion to ortho-phosphorus using acid hydrolysis. The Skalar autoanalyzer system
is used for inline hydrolysis using sulfuric acid added to the sample stream and heated
to 97C. Colorimetric determination follows method WP04, so WHPO4 includes
reactive phosphorus.

WTOTP: Total Phosphorus (UV Digestible)

For the determination of dissolved total phosphorus, filtration of the sample through a
0.45mm filter is performed either in the lab or in the field. Following hydrolysis
(WHPO4), the sample undergoes further digestion with peroxodisulfate under UV
radiation. Colorimetric determination follows method WPO4.

LABORATORY 7: Chlorophyll A
Trichromatic Method for Chlorophyll Analysis: (Strickland and Parsons,
1968)
There are several methods of obtaining an estimate of primary productivity. The
four most significant methods are counts, dry weights, turbidity and chlorophyll analysis.
Each of these methods has advantages and disadvantages. Counts are generally
considered most satisfactory since only with this method are the species of algae
actually differentiated. However, the method is very slow and a great deal of the
accuracy depends on the operator. Dry weights and turbidity are both very fast
methods but it is difficult to distinguish the algae from the debris. Chlorophyll analysis
has the advantage of being both fast and specific. Since it measures a component of
the living cytoplasm, it gives an idea of the potential for growth of the algae.
Unfortunately, it is difficult to correlate chlorophyll determinations with dry weight
measurements or aerial standard unit counts or other assessments of productivity.

Spectrophotometric Determination of Chlorophyll a, b, c, and Plant

Carotenoids:
The plant pigments of algae consist of the chlorophylls and carotenoids
(carotenes and xanthophylls).
The three major chlorophylls, a, b, and c absorb light
maximally at specific wavelengths when dissolved in organic solvents. From these
absorption characteristics, an estimate can be made of the concentrations of the
pigments. Chlorophyll a is by far the most dominant chlorophyllous pigment and occurs
in greatest abundance. Therefore, chlorophyll a alone can be used to estimate algal
biomass. The spectrophotometric estimate of chlorophyll c concentration in the
trichromatic method below is only approximate. Using a more elaborate extraction
method yields higher precision measurements of chlorophyll c (see Strickland and
Parsons, 1968); Chlorophyll concentrations are expressed in g/L (or mg/m3).
Estimates of plant carotenoid pigments are reported collectively in pigment units that
approximate mg/m3 (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 relatively
constant relationship exists between chlorophyll and photosynthesis at any given light
intensity. Chlorophyll a is the master pigment in blue-green and eukaryote
photosynthesis. In a living cell, chlorophyll a absorbs light in two peaks - one between
670 and 680 nm and the other at 435 nm. The longer wavelengths (670 680 nm) are
abundantly present in shallow water while the shorter wavelengths penetrate deeper,
thus allowing photosynthesis to occur at many levels (Cole 1975). Since plants contain
assortments of accessory pigments. Other light waves that travel vertically in the lake
also play a part in photosynthesis. Accessory pigment molecules absorb energy quanta
from light waves, become exerted' and energy-rich, and pass their excitation energy on

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

APPARATUS:
25 mL graduated cylinder
10 mL graduated cylinder
Plastic filtration apparatus w/hose
Plastic filter membranes
Homogenizer unit with homogenizer tubes (Pyrex)
Pipettes
10 mL graduated centrifuge tubes
Centrifuge
Spectrophotometer
Quartz 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 one
circle of the plastic membrane on the filter base. DO NOT touch the plastic
membrane with your fingers; it will disintegrate.
2) Connect the filtration unit to the vacuum and open the vacuum very slowly and
slightly.
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 and
allow the filter to dry (about one minute).
6) Using the tweezers, remove the plastic membrane from the filter base and place
the membrane in a homogenizer tube. Place the membrane as close to the
bottom of the tube as possible.
7) Add 8 ml of 90% acetone to the homogenizer tube and macerate the membrane
completely using the homogenizer. In the process, the algal cells are dissociated

and the pigments extracted. When maceration is complete, transfer the contents
to a 15 mL graduated centrifuge tube.
8) Rinse the homogenizer tube with 4 mL of 90% acetone and transfer the contents
to the same centrifuge tube.
9) Centrifuge the tube at approx. 2500 rpm (speed setting 1-2) until clear, about 20
minutes.
10) Using a pipette, transfer about 5 mL of the supernatant to a quartz
spectrophotometer cuvette and read the absorbance at 663, 645 and 630 nm on
the spectrophotometer in the instrumentation laboratory.
11) Record your observations and calculate the Chlorophyll a concentration in terms
of mg CHa/L using the following equations:
Acorr = An - A750 (turbidity correction)
Correct for turbidity for each absorbance value first and then put the corrected
values into the following equation:
A = (11.64 A663 - 2.16 A645 + 0.1 A630) x 12.0 mL /0.025 L
For your report calculate the mg CHa/L for each groups results and explain any
differences observed. In addition, give a brief discussion of the
advantages
and disadvantages of this technique

LABORATORY 8: DETERMINATION OF HEAT BUDGET FOR

MIRROR LAKE
One of the most important and interesting characteristics of a lake is the thermal
structure. 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 the
growth and distribution of plants, animals and microbes (Wetzel and Likens 1979).
Large volumes of water change temperature relatively slowly because of the high
specific heat of water. Therefore, large lakes tend to moderate local climates and
provide longer growing seasons for aquatic life, and serve as integrated recorders of
recent climatic phenomena. These are some of the reasons that the thermal structure
and heat content of a body of water must be known, with some degree of accuracy, in
limnological studies.
Determining a heat budget for 'Mirror Lake' requires a calculation of the heat
content 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 determined
from:
zm
w = tz Az hz
zo
w = heat content of the lake water in calories
zo = surface of the lake
zm = maximum depth of the lake
tz = average temperature in C of a unit layer of water of thickness hz (in cm, with the midpoint at
depth z)
Az = the area at depth z in cm2
(The heat content is usually expressed on a unit area basis, w/Ao in cal/cm2 where Ao = surface
area in cm2)

Since the specific heat of water is one cal g-1C-1 and since one cm3 of water has a
mass of about one g, it is convenient to use a constant area of one cm2 in calculations
of heat content for water, i.e. Az = 1 cm2. However, because most natural lakes do not
have basins with perpendicular walls, it is necessary to correct for heat content that
varies with depth. This may be done by using a ratio of the area at depth z to the area
of the surface of the lake. The ratio is usually obtained from a hypsographic or
hypsometric curve.
Use the Sample data sheet below to calculate the heat content of Minor Lake for the
two dates. Answer the questions below:

Using the data provided for the dates specified, calculate a heat budget for the period
between the May and June dates. Did Mirror Lake lose or gain heat over this period?
Cole, Gerald A., 1994, Textbook of Limnology, 4th Edition, Waveland Press, Inc.,
Prospect Heights, I1., p.233.
Wetzel, Robert G. and Likens, Gene E., 1979, Limnological Analyses, W.B. Saunders
Company, Philadelphia, Pa., p. 46, 51, 53-55.

Sample Data Sheet for the Heat Content Calculation:

1
Reference
Depth (m)

2
Thickness of
Layer (cm)

3
Weighting
Factor from
Hypsographic
Curve
(% expressed
as decimal
fraction)
"For Mirror
Lake
calculations
use Table 4-2
area %

4
Average
Temperature
of Layer (C)
Must be
calculated
as
T
1+T2/2
(Temp. at
each end of
the
stratum)

5
Caloric
(heat)
Content

(Temp. x
Thickness)
[(2) x (4 )]

6
Weighted
Caloric (heat)
Content/Layer
(cal/cm2 of
lake surface)
[(3) x (5)]

* Total Column #6 to obtain the caloric (heat) content of the entire lake in cal/cm2
Calculate the total heat content of Mirror Lake for the 17th of May 1970 and 24th of June 1970.
What was the heat budget for Mirror Lake (i.e. the difference in heat content over some time
interval) over this interval? Did Mirror Lake gain or lose heat over this period?

TABLE 4-1

Temperature profiles and other physical characteristics in Mirror Lake, New Hampshire.

Depth (m)
0
0.51
0.62
1
2
3
4
5
6
7
8
9
10
10.3 (mud surface)
Ice thickness (cm)
Snow depth (cm)
Water level
depression below
ice surface (cm)
Air temp. (C)

17 May
1970

24 June
1970

Temp. (C)
15.45
15.45
13.58
12.15
9.73
8.46
7.06
6.20
6.06
5.45
5.62
5.75

Temp. (C)
21.80
21.35
20.74
20.41
16.53
12.38
10.27
8.69
7.38
6.52
6.48
6.50

12.1

22.2

TABLE 4-2 Hydrographic data for Mirror Lake, New Hampshire.

Location 43 56.5'N, 71 41.5'W
Elevation (m) 213
Maximum depth (m) 11.0
Surface area (ha) 15.0
Mean depth (m) 5.75
Drainage area (ha) 85
Relative depth (%) 2.5

Depth
0
1
2
3
4
5
6
7
8
9
10
10.9

Area
m2 x 104
15.0
13.6
12.5
11.5
10.6
9.9
9.1
7.0
3.2
1.6
0.6
0
Total

%
100
90.7
83.3
76.7
70.7
66.0
60.7
46.7
21.3
10.7
4.0
0

Depth
0-1
1-2
2-3
3-4
4-5
5-6
6-7
7-8
8-9
9-10
10-10.9

0-10.9

Volume
m3 x 103
143
130
120
110
102
95.0
80.3
49.8
23.5
10.6
2.00

866.2

%
16.5
15.0
13.9
12.7
11.8
11.0
9.3
5.7
2.7
1.2
0.2

100.0

From: Wetzel, Robert G. and Likens, Gene E., 1979, Limnological Analyses, W.B. Saunders Company,
Philadelphia, Pa., p. 46, 51, 53-55.

LABORATORY 9: WATER QUALITY ASSESSMENT

One of the many effects that organic wastes have on the aquatic environment is
to 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, or
pollution. 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 the
rate of oxygen removal by bacteria and protozoa is high. Algal species tolerant of
organic pollution are significant in the recovery of a stream or lake because they add
oxygen to the water during photosynthesis and incorporate organic and inorganic
nutrients into their cells, thus removing pollutants from the water.
Algae is often used as a biological indicator species in determining the type and
degree of pollution m a body of water Three concentrated water samples will be
analyzed 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 concentrate
allows the researcher to observe a large number of algae in a small sample). The
objective is to count each of the algae in a known volume of sample, and then apply
Palmer'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. You
should be familiar with all of them before leaving the lab today (Oil immersion will not be
necessary for our purposes).
You will need:
One microscope
Three microscope slides
Three cover slips (25 mm x 25 mm)
A keen eye
Lots 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 the
cover slip (Place the pipette as close to the bottom as possible). Try to avoid the large
green masses of algae, as you will have a difficult time identifying individual species in
these clumps. Air bubbles are not allowed either.

4) At your desk, view the sample at powers 10 and 45. At power 10, the diameter of
your field is roughly 2000 micrometers (2 mm); at power 45, it is about 500
micrometers. Use these facts to help you identify algae samples according to size (see
picture key to Algal Genera).
5) When you find a species, record that you have done so in the appropriate chart. Not
all species occur in all samples.
6) It would be tedious to count the entire contents of your 0.1 mL sample. Three "strips"
of the sample will be counted instead. Each strip will consist of the length of your cover
slip (25 mm) and the diameter of the microscope field at power 10 (not 45).
7) For each strip, count and record the number of each genus. Colonies and or
filaments are counted as a unit. Large filaments or colonies that are only partially lying
in the strip should be counted as fractions.
When you have completed this part of the exercise, record your findings on the master
chart at the front of the lab. When all groups have reported results, you may proceed
with the rest of the lab.
Copy the group results onto the chart provided. Calculate the totals where indicated.
Use the group results in the following exercise.
For each of the three water samples:
1) Calculate the density (number of algal units per mL) for each algal genus
identified. Use the following formula to calculate algal units per mL of
concentrated sample. Record your results on Chart 2.
Note: If the total for any of the algal genera in any sample is less than 20, do not
use those Genera in your calculations.
(AREA OF COVER SLIP) x (NUMBER OF UNITS FOR ONE ALGAL GENERA
(AREA OF ONE STRIP) x (NUMBER OF STRIPS COUNTED) x (VOLUME UNDER SLIP)

*Important* The number of strips counted refers to the total number counted by
all the groups for that genera. E.g. If there are 10 groups reporting results, # of
strips = 10 x 3= 30
2) Calculate the number of organisms in the actual lake water sample (i.e. before
the water was filtered).
# OF ALGAE PER ML CONCENTRATED SAMPLE x TOTAL VOLUME OF CONCENTRATE
TOTAL VOLUME OF LAKE WATER FILTERED TO YIELD CONCENTRATE

3) Determine the Pollution Index Value for each water sample. Palmer (19139)
identified a list of organic pollution tolerant algae, to which he assigned Pollution
Index Values (Table 1). The Index can only be applied to those genera m a
given sample that have a concentration of > 50 individuals per mL of lake water
sample (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 (<15) indicate the absence of
organic pollution.
4) Answer the questions assigned. Hand in the entire lab by the end of the next lab
period
Worksheets are in Appendix 2
Literature Cited:
Palmer, C. Mervin. 1969. A composite rating of algae tolerating organic pollution.
Journal of Phycology. 5 (1): 7882
Questions:
1 According to the Pollution Index, what sample was the most polluted? The
least polluted?
2. Which sample had the highest diversity of organisms? The least diversity?
3. Which bottled sample was the most turbid? The least turbid?
4. What would you expect the colour of water to be in a highly organically
polluted lake?
5. What algal species (studied in the lab) would you expect to find in the greatest
numbers in polluted waters?

Testing of Environmental Samples Using the LC50 Quantal

Test
Toxicology studies, routinely carried out in the field of Limnology, are becoming
increasingly important for environmental protection. This lab exercise will introduce you
to the LC50 test, a quantal test to determine the Lethal Concentration causing death in
50% of the test subjects, using neonate Daphnia spp. (<24 hours old). The class will
head down to the Aquatic Toxicology Research Centre (ATRC) on the ground floor of
the Centennial Building (CB0020). The technicians in the toxicology lab will discuss
with you the LC50 test and will demonstrate the techniques involved.
For your information, the standard operating procedure (SOP) for this particular
test will be provided for you to review. You will recall from the guest presentation, that
the SOP is essential in assuring quality control in your test results. This SOP is what
the technician will be following with you.
The following background information will assist you in your understanding of an
LC50 test. This information has been taken from Environment Canada's Guidance
Document on Application and interpretation of Single-species Tests in
Environmental Toxicology. EPS 1/RM/34-December 1999.
LC50 is the median lethal concentration, i.e. the concentration of material in air, water,
soil, or sediment that kills 50% of the test organisms. The LC50, and its 95%
confidence limits, are statistical analyses of the percent mortalities from several test
concentrations with a fixed exposure time. The duration of exposure is always reported
along with the LC50 value, e.g. 48-h LC50.
Materials Tested:
Toxicity of environmental samples is often assessed by the effect of full-strength
material, as an alternative to estimating the percent of full strength material that is
needed to produce a defined effect such as 50% mortality. The concentration or even
the identity of the toxicant(s) producing the effect might remain unknown. For
environmental samples, the upper limit for concentration is full strength. If that has no
effect, the sample can only be characterized as non-toxic.
In a quantal test, each organism either shows the effect or does not. The endpoint is
the concentration causing an effect in the median organism, the LC50, or the EC50, i.e.
median effect concentration - conc. of material in water, soil, or sediment that causes a
defined effect to 50% of the test organisms.

Quantal Tests:
The example of a concentration-effect curve (or concentration-response curve) in
Figure 1 represents results from an aquatic toxicology test. Figure l relates the intensity
of 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 the
middle of the percent scale, since such values have the least variability and the most
value or "weight" in estimating the nearby endpoint for 50% effect. There should also
be a pronounced gradient of effect, ranging from little or no effect at a low concentration
to complete or nearly complete effect at a high concentration. Those very low and very
high percent effects have reduced weight in fitting the line, but they help to anchor it and
establish the slope, which is important in calculating confidence limits.

Figure 1

Results of a LC50 Test Plotted on a Logarithmic-probability Scale. A straight line

has been fitted to the data. The LC50 has been estimated graphically as about 5.5 mg/L
by the line drawn across from 50% effect, then vertically down from the intersection with
the fitted line. Probit analysis on computer confirmed the LC50 as 5.6% and estimated
the 95% confidence limits shown by the horizontal bar at 50% effect.

The median effect has the greatest precision as an endpoint for quantal tests.
Familiar examples are the median lethal concentrations (LC50) and the median
effective concentration (EC50). Half of the test organisms would have shown the effect
at concentrations lower than the endpoint (C). The other half of the organisms would
only 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 effect
being tabulated must also be stated.
In quantal experiments, the endpoint is estimated by fitting a line to the
concentration-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 other
specialized 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 probit
line. The mathematical model of the concentration-effect relationship also describes the
associated error term and estimates the precision of the endpoint, customarily
expressed 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 be
those of all acute lethality tests, a series of tests with different exposure times would be
expected to result in a series of different probit lines. Short exposures would require
higher concentrations to manifest mortalities from 0 - 100%. Long exposures would be
expected to require lower concentrations. However, in acute testing, there is often an
exposure 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.

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 their
behaviour in the environment and organisms.
3. What is an LC50?
4. Explain the difference between a high Kow and a low Kow.
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 used
rather than dose vs. response in determining an LC50 or LD50.

Appendix 1
Writing a Formal Report or Scientific Article in Science

Sections:
Title
The title of a report/article is the first part of your report that will be read and
possibly the only part read for journal articles. Therefore, the title must serve two
purposes; a title informs the reader about the subject of the study, and
distinguishes your work from similar studies in the literature.
Titles are not necessarily complete sentences; titles are pared down to contain
only the words essential to conveying the important details. There should be a
clear relationship between all of the details mentioned in the title, and although
the title should distinguish your work from others, trying to cram too much detail
into a title will make unreadable. Browsing scientific journals especially those
in 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 following
information 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 not
include references or citations in an abstract and only include information found
in the body of the report. Readers of scientific journals will typically only read the
title and the abstract if the title was appealing enough , so a good abstract is
important for getting your article read.

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

introduction acronyms used should be written out in full followed by the

acronym 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 fewer
references than is appropriate, but it is a writing skill that will improve with
experience more discussed in the References section.
The length of the introduction will vary with the material introduced; the
introduction should be concise, but should be long enough to fully prepare the
reader for the reported study.

Results and Discussion

The results and discussion of a report/article are where you get to present your
findings. The previous sections of the report prepare the reader for what you
show them in this section; the readers are now familiar with the background and
subject of the study, the techniques used, and why the study was performed. All
that remains for this section of the report is for you to point out the important
findings and discuss their significance within the scope of the study.
There are two common styles for this section; the results and discussion can be
combined 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 they
are 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 must
be guided through the results section and directed to the important details in the
illustrations - 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, and
the implications of the findings when interpretation is done.
Interpretation includes discussing the significance and implications of the results
within 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 studies
need to be explained - . Do not make generalized statements that are not based
on your data or known fact, however.
There is some choice for how the findings are presented; results can be part of
the text, displayed in tabular form, or compiled into a figure/graph. The
presentation choice is determined by what you are trying to say about the data
and how appropriately the text/table/figure gets your point across. All

illustrations figures and tables must be introduced in the text before they are
included in the report, to prevent confusing the reader.
Illustrations can either be embedded in the text between paragraphs or on
separate pages; the number of illustrations per page will be determined by the
illustration size and the order of introduction in the text. Embedded illustrations
are inserted immediately following the paragraph they are introduced in. If
placing illustrations on separate pages, place them at the end of your report,
following Literature Cited the results must contain text introducing and referring
to the these figures.
Each illustration must be able to stand alone, the reader should not need the
text to understand what the illustration shows. To stand alone, illustrations need
to have all units labelled, clearly labelled axes or column titles, an appropriate
scale if graphing and a descriptive title. Be sure to include the most
important detail from the illustration in the title. Figure titles are placed at the
bottom of the figure, whereas table titles are placed at the top of the table.
There must be a summary of what you concluded from the experiment, but no
separate conclusions section. Generally, the conclusion restates the
objectives, summarizes the important results and restates the major conclusions
drawn from the results, and makes some consideration for future work areas
that should be studied to expand or clarify parts of the study, and improvements
for later studies.

Literature Cited
This is an alphabetical listing (by senior author's surname) of all the sources cited
in the text of your paper. Note that the sources here must match those in the text
of your report. For example, if no references appear in your paper, this section
would be empty and you would have a very short paper. Do not number the
references. Use a hanging paragraph format for references in this course
other courses may require a different format and journals each have their own
preference for reference format and style.
If you are citing a book, use this format:
Brnmark, C. and L.-A. Hansson. 2005. The Biology of Lakes and Ponds. 2nd ed.
Oxford University Press Inc. New York: 285 pp.
If you are citing a journal article, such as Alces, use this format:
Hicks, A. 1986. The history and current status of moose in New York. Alces. 22:
245-252.

Do not use footnotes; place your references in the literature cited section. In
scientific papers, full quotes are not used; material is paraphrased and a
reference is cited at the appropriate place. Citations are usually placed at the
end of the sentence, but if several bits of information are used from the same
source, 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 by
Smith et al. (2003) that the sky is blue.
Note: For a book, the total number of pages is listed, but for a journal article the
article page numbers are listed.

Why Use References?

A common problem with student papers is plagiarism, i.e. presenting the ideas
and findings of others as if they were your own without credit/references. Usually
this 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 a
research career prior to the course , you must give credit to the authors.
Plagiarism also applies to copying another students paper and submitting it as
your own. Additionally, you cannot resubmit a paper you wrote for another course
or in another year.
Plagiarism is a serious academic offence. Plagiarism is defined in University
Regulation IX Academic Dishonesty as:
1. Plagiarism of ideas as where an idea of an author or speaker is incorporated
into the body of an assignment as though it were the writer's idea, i.e. no credit is
given the person through referencing or footnoting or endnoting.
2. Plagiarism of words occurs when phrases, sentences, tables or illustrations of
an author or speaker are incorporated into the body of a writer's own, i.e. no
quotations or indentations (depending on the format followed) are present but
referencing or footnoting or endnoting is given.
3. Plagiarism of ideas and words as where words and an idea(s) of an author or
speaker 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 or
endnoting is given.
From University Regulations IX Academic Dishonesty
http://calendar.lakeheadu.ca/current/contents/regulations/univregsIXacdishon.html

Additionally, plagiarism is considered misconduct under Code of Student

Conduct and Disciplinary Procedures (http://policies.lakeheadu.ca/policy.php?pid=60).

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 in
expulsion from the University and a mark of zero for the course a mark that is
often 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. Genus is Capitalized, species is not.
7. Check for correct use of affect/effect.
8. Also check for their/there/theyre.
9. And for Its (It is, It has)/Its.
10. And for lose/loose (trust me; it happens more than you think).
11. Dont use contractions.
12. Proof-Read!

Marking
The marking scheme for reports will be as follows:

Appendix 2
Worksheets for Laboratory 9

NAME:___________________________________________

NAME OF PARTNER:_______________________________

SAMPLE NUMBER:_________________________________

ALGAL SPECIES
CLOSTERIUM
SYNEDRA
MICRASTERIAS
CHLORELLA
STAUSTRUM
PHACUS
EUGLENA
CHLAMYDOMONAS
PEDIASTRUM
SCENEDESMUS
PANDORINA
OSCILLATORIA
BULBOCHAETE
STIGEOCLONIUM

FOUND

NOT FOUND

NAME:___________________________________________

NAME OF PARTNER:_______________________________

SAMPLE NUMBER:_________________________________

NUMBER
ALGAL SPECIES
STRIP 1
CLOSTERIUM
SYNEDRA
MICRASTERIAS
CHLORELLA
STAUSTRUM
PHACUS
EUGLENA
CHLAMYDOMONAS
PEDIASTRUM
SCENEDESMUS
PANDORINA
OSCILLATORIA
BULBOCHAETE
STIGEOCLONIUM

STRIP 2

STRIP 3

STRIP 4

NAME:___________________________________________

NAME OF PARTNER:_______________________________

SAMPLE NUMBER:_________________________________

ALGAL SPECIES
CLOSTERIUM
SYNEDRA
MICRASTERIAS
CHLORELLA
STAUSTRUM
PHACUS
EUGLENA
CHLAMYDOMONAS
PEDIASTRUM
SCENEDESMUS
PANDORINA
OSCILLATORIA
BULBOCHAETE
STIGEOCLONIUM

FOUND

NOT FOUND

NAME:___________________________________________

NAME OF PARTNER:_______________________________

SAMPLE NUMBER:_________________________________

NUMBER
ALGAL SPECIES
STRIP 1
CLOSTERIUM
SYNEDRA
MICRASTERIAS
CHLORELLA
STAUSTRUM
PHACUS
EUGLENA
CHLAMYDOMONAS
PEDIASTRUM
SCENEDESMUS
PANDORINA
OSCILLATORIA
BULBOCHAETE
STIGEOCLONIUM

STRIP 2

STRIP 3

STRIP 4

NAME:___________________________________________

NAME OF PARTNER:_______________________________

SAMPLE NUMBER:_________________________________

ALGAL SPECIES
CLOSTERIUM
SYNEDRA
MICRASTERIAS
CHLORELLA
STAUSTRUM
PHACUS
EUGLENA
CHLAMYDOMONAS
PEDIASTRUM
SCENEDESMUS
PANDORINA
OSCILLATORIA
BULBOCHAETE
STIGEOCLONIUM

FOUND

NOT FOUND

NAME:___________________________________________

NAME OF PARTNER:_______________________________

SAMPLE NUMBER:_________________________________

NUMBER
ALGAL SPECIES
STRIP 1
CLOSTERIUM
SYNEDRA
MICRASTERIAS
CHLORELLA
STAUSTRUM
PHACUS
EUGLENA
CHLAMYDOMONAS
PEDIASTRUM
SCENEDESMUS
PANDORINA
OSCILLATORIA
BULBOCHAETE
STIGEOCLONIUM

STRIP 2

STRIP 3

STRIP 4

TABLE 1
Algal Genera Pollution Index (Palmer, 1969)
Genus
Anacystis
Ankistrodesmus
Chlamydomonas
Chlorella
Cyclotella
Euglena
Gomphonema
Lepocinclis
Melosira
Micractinium
Navicula
Nitzshia
Oscillatoria
Pandorina
Phacus
Phormidium
Scenedesmus
Stigeoclonium
Synedra

Pollution Index
1
2
4
3
1
5
1
1
1
1
3
3
5
1
2
1
4
2
2