The procedures of this laboratory activity are adapted from the article Mehmood Et Al, S. (2021).

Length-weight relationship,
morphometric and meristic controlling elements of three freshwater fish species inhabiting North-Western
Himalaya. Egyptian Journal of Aquatic Biology and Fisheries, 25(6), 243-257.

CHEMBIO 2 Analytical Methods in Biology

Biodiversity and Morphometric Analysis

Introduction

Knowing what species inhabit an ecosystem, and how many of each kind there are, is critical to
understanding that ecosystem's structure and function, and predicting future changes. Scientists
who look at the variation of life in a forest, a stream or a patch of soil are looking at its biodiversity.
A healthy ecosystem is a balanced one—with enough predators, prey, producers and
decomposers to keep the food web stable. When the system is thrown off balance, that web can
quickly unravel. Forests may struggle to grow when the deer population explodes, or fish near the
top of the food chain may suffer when tiny organisms at the base disappear. But when biodiversity
levels are right, it can strengthen the ecosystem to better resist stresses like climate change and
invasive species.1

And sampling of macroorganisms entails the examination of their morphology. The study of
morphology is a common means of biological grouping and classification.2 Scientists use
morphometric analysis. Morphometric techniques aim at measuring size, shape, and the relation
between size and shape (allometry). This will aid in the classification of organisms and how their
environment influences their physical characteristics implying adaptation.

In this activity, you will apply the methods of biodiversity assessment and examine the anatomy
of representative organisms by morphometric analysis and dissection.

Materials (per group): • Ruler

• Tape measure • Pins and small hammer
• Marking materials (for the plot) • Old newspapers
• Thermometer • Cleaning materials
• 1 Cotton ball
• 1 Rubber band Specimen (per group):
• Paper and pen (for recording) • 2 mussels
• Dissecting pan • 1 snail
• Dissecting tools (cutter, too) • 1 squid
• Digital weighing balance • 1 fish
• Magnifying glass

Procedures:

PART 1 ECOLOGICAL ASSESSMENT

1. Set up a sampling plot measuring 3m x 3m. Obtain the GPS location of your area.
2. Record the following abiotic factors:
a. Temperature
b. Humidity
i. Get the dry temperature by recording the overall temperature.
ii. Obtain the wet temperature by wetting a cotton ball and attaching this at the tip of a
thermometer by a rubber band.
iii. Calculate humidity using the formula:
wet temperature
x 100
dry temperature
3. Describe the sampling area in terms of:
a. Presence of a water body
b. Vegetation
c. Presence of infrastructures/overall location
4. Document your area by taking a photograph.

1
https://serc.si.edu/research/research-topics/biodiversity-conservation/biodiversity-assessment
2
Doyle, D., Gammell, M. P., & Nash, R. (2018). Morphometric methods for the analysis and classification of gastropods: a comparison using Littorina
littorea. Journal of Molluscan Studies, 84(2), 190-197.
 5. Using your pen and paper, record all the different plant and animals that you can observe in your area.
Describe each organism. Categorize into plants and animals.
6. Obtain the population of each organism you recorded.
7. Calculate following diversity indices:
a. Species richness and species evenness
b. Simpson’s index
8. Obtain a sample of 3 plants and 3 animals you have recorded for the morphometric examination.
9. Clean up the area you used for the ecological assessment.

PART 2 MORPHOMETRIC ASSESSMENT AND DISSECTION

1. Make sure that you accomplish routine pre-lab procedures before starting.
a. Complete lab gear and materials.
b. Arrange materials on workstation.
c. Clean station and materials.
2. PART 2A Morphometric assessment of specimens from PART 1
a. Categorize your samples from PART 1.
i. Classify your plant specimens. See figure below.

Google images

ii. Use the following leaf morphometric parameters. Measure using cm:
• number of leaves
• margin of leaf
• leaf length (Ll)
• leaf width (Lw)
• petiole length (Pl)
• leaf base width on 1 cm from petiole base (Blw)
• number of veins – left (Vl)
• number of veins – right (Vr)
• distance between 3rd and 4th vein - left (Dv 3-4), see Figure 1.

Šijačić-Nikolić, M.,
Milovanović, J., Nonic, M.,
Knezević, R., & Stanković, D.
(2013). Leaf morphometric
characteristics variability of
different beech provenances in
juvenile development
stage. Genetika-
Belgrade, 45(2), 369-380.
 iii. Morphometric analysis of specimens obtained from your ecological assessment:
• Overall body color/pigmentation
• Number of segments/tagmata
• Number of legs
• Presence of antenna
• Type of mouthparts
• Presence of wings
• Number of wings
• Mean length of wings
• Length of body

3. PART 2B Morphometric assessment and dissection of specimen you brought

4. Mussel
a. Morphometry. Measure the shell in 3 parts in centimeters:
i. from the anterior to the posterior end (length)
ii. from the dorsal to the ventral end (width)
iii. height

iv. Measure the weight of each

mussel in 2 ways:
A. Intact (whole
specimen)
B. Both shells
only (without
the innards)
v. Count the annual growth
rings. These growth rings
are the dark bands in the
external of the shell. Count
the darkest rings, each ring
represents 1 year in clams
but in mussels, 1 growth ring
is equivalent to 1 month.
See the figure on the right.

b. Dissection
i. Place a mussel in a dissecting tray and identify the anterior and posterior ends of the
mussel as well as the dorsal, ventral, & lateral surfaces.

ii. Turn the mussel with its dorsal side down and insert the scalpel between the ventral edges
of the valves and force open the mussel. Carefully work the tip of the scalpel between
the valves so you do not jab your hand.
iii. Observe the internal anatomy. Refer to the figures below.
5. Snail.
a. Morphometry
i. Measure the following from the specimen in cm:
• Shell height
• Shell width
• Number of whorls
• Describe the shell color
ii. Observe the external anatomy of the snail specimens by referring to the figure below.

b. Dissection
i. Using a small hammer, crack the shell with ample force that will not squish the entire snail.
ii. Obtain the internal structures and wash away the shell particles using the wash bottle.
iii. Observe the anatomical structures.
 iv. Cut along the length of the thin membrane to expose all organs.
v. Cut along the broken lines as indicated in the figure below on the left with black broken
lines. Avoid all internal organs/structures by only cutting the thin (transparent)
membrane. Continue with the incision along the edge of the membrane all the way to the
first whorl. This will expose portions of the reproductive and digestive system. Also, cut
along the lines indicated in the last figure with black broken lines to expose the base of
both systems.

vi. Peel back the membrane to expose the internal organs

vii. Peel back the transparent membrane to expose the internal organs. Continue with the
incision made in the last figure above all the way to the end of the coiled regions of the
animal (portion that was retained inside the shell). The animal may be inverted to
accomplish this as indicated by the broken lines in the figure on the left.
viii. Observe the rest of the organs
 ix. Remove the ovotestis from digestive gland
x. Slowly tease the ovotestis and the albumen gland away from the digestive gland. Both
organs can be carefully separated with a pair of tweezers. Once dislodged, both systems
can be separated as indicated in the figure below. You may detach several systems of the
snail.

6. Squid dissection.
a. Morphometry
• Length of trunk
• Length from tip of trunk to the longest arm
• Length from tip of trunk to the longest tentacle
• Mean length of arms
• Mean length of tentacles
• Mean diameter of fins
• Width of median trunk
• Weight

o Dissection
i. Before making incisions in the squid, observe the dark spots. What do you think are
these?
 ii. Spread the tentacles and arms apart to find the squid's mouth. With the tweezers, you
can pull out the beak from the mouth enclosure called the buccal mass. When it comes
out without breakage, it looks a lot like a parrot's beak. What do you think is this for?

iii. Place the squid in a dissecting tray and

identify
the anterior and posterior ends of the
mussel as well as the dorsal, ventral, &
lateral surfaces.
iv. Make sure that the arms are on the
bottom part of the dissecting tray.
v. Look for the siphon. With a scalpel,
lightly cut apart the mantle in the middle
so as not to cut the internal organs.
Spread out the mantle flat on the tray
and see what's inside.
vi. Identify if your squid is male or female.

7. Fish morphometry.
a. Below are illustrations obtained from the paper of Zafar, M., Nazir, A., Akhtar, N., Naqvi, S. M., &
Zia-ur-Rehman, M. (2002). Studies on meristic counts and morphometric measurements of
mahseer (Tor putitora) from a spawning ground of Himalayan foot-hill river Korang Islamabad,
Pakistan.
b. Obtain the following meristic count (number) parameters using the illustration below as guide.
• dorsal fin rays,
• pectoral fin rays,
• pelvic fin rays,
• anal fin rays,
• caudal fin rays,
• lateral line scales,
• scales above the lateral line and
• scales below lateral line
 c. Measure the morphometric parameters using the illustration below as guide.
• Weight • 1st dorsal fin base
• Total length • Interdorsal space
• Standard length • 2nd dorsal fin base
• Fork length • Depth of 2nd dorsal fin
• Head length • Length of caudal peduncle
• Postorbital length • Depth of body
• Eye diameter • Length of pectoral fin
• Preorbital length • Anal fin base

4. Complete the tables on the next page.

5. Document your dissection activity. Submit via MSTeams.
6. Supply the following data in the following tables with your recorded data:
a. Ecological assessment
b. Morphometric assessment of collected specimens
i. Plants
ii. Animals
c. Meristic and morphometric assessment
i. Mussel
ii. Snail
iii. Squid
iv. Fish
7. Answer the following questions:
a. Based on the ecological assessment you conducted in your sampling area, what conclusion can
you generate? Why is assessment and measurement in Ecology important?
b. Why is morphometry important in observing representative organisms? Dissection?
c. What generalizations/insight can you obtain from this dissection activity? Were you able to gain
important skills from the exposure to relevant hands-on activities during the semester?
8. Submit. DEADLINE: January 27, 2023
 CHEMBIO 2 Analytical Methods in Biology

Biodiversity and Morphometric Analysis

Results and Discussions

A. Ecological Assessment

Picture of sampling area

General location of the sampling area: ____________________________________________________

GPS location: _________________________________________
Abiotic factors:
• Temperature _________________________
• Humidity _________________________
Geographic description:
a. Presence of a water body
________________________________________________________________________________________
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b. Vegetation
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c. Presence of infrastructures/overall location
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________________________________________________________________________________________

Population and diversity assessment

Table 1. Plants
Number of Species
Species
Description individuals evenness (in
code
percentage)
 Table 2. Animals
Number of Species
Species
Description individuals evenness (in
code
percentage)

Species richness: _________________

Simpson’s diversity index with description: _________________
________________________________________________________________________________________
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B. Morphometric Analysis of Collected Specimens (May use a separate paper)

Table 3 Plants
Parameters Specimen 1 Specimen 2 Specimen 3
Classification

Number of leaves
Margin of leaf
leaf length (Ll)
leaf width (Lw)
petiole length (Pl)
Morphometry

leaf base width on

1 cm from petiole
base (Blw)
number of veins –
left (Vl)
number of veins –
right (Vr)
distance between
3rd and 4th vein

Table 4 Animals
Morphometry Specimen 1 Specimen 2 Specimen 3
Classification
presence of vertebrae
Phylum
Overall body
color/pigmentation
Number of
segments/tagmata
Number of legs
Presence of antenna
Type of mouthparts
Presence of wings
Number of wings
Mean length of wings
Length of body
C. Meristic and Morphometric Analysis of Obtained Specimens

Mussel
Parameters Mussel 1 Mussel 2
Classification (presence of vertebrae)
Phylum
Length
Width
Height
Weight (whole specimen)
Weight (both shells only)
Number of growth rings/lines

Snail
Parameters Snail specimen
Classification (presence of vertebrae)
Phylum
Shell height
Shell width
Number of whorls
Describe the shell color

Squid
Parameters Squid specimen
Classification (presence of vertebrae)
Phylum
Length of trunk
Length (tip of trunk to the longest arm)
Length (tip of trunk to the longest
tentacle)
Mean length of arms
Mean length of tentacles
Mean diameter of fins
Width of median trunk

Fish (Meristics)
Parameters Fish specimen
Classification (presence of vertebrae)
Phylum
Dorsal fin rays,
Pectoral fin rays,
Pelvic fin rays,
Anal fin rays,
Caudal fin rays,
Lateral line scales,
Scales above the lateral line and
Scales below lateral line

Fish (Morphometrics)
Parameters Fish specimen
Weight
Total length
Standard length
Fork length
Head length
Postorbital length
Eye diameter
Preorbital length
1st dorsal fin base
Interdorsal space
2nd dorsal fin base
Depth of 2nd dorsal fin
Length of caudal peduncle
Depth of body
Length of pectoral fin
Anal fin base
D. Based on the ecological assessment you conducted in your sampling area, what conclusion can you
generate? Why is assessment and measurement in Ecology important?
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E. Why is morphometry important in observing representative organisms? Dissection?

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F. What generalizations/insight can you obtain from this dissection activity? Were you able to gain important
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I am enough of an artist to draw freely upon my imagination. Imagination is more important than knowledge.
Knowledge is limited. Imagination encircles the world.
~ Albert Einstein