Faculty of Applied Sciences

LABORATORY MANUAL

Subject Code : MS010 / DAA2074

Subject Name : Aquatic Health and Diseases

Semester : May 2024

Lecturer :
Practical Practical title Page Date
1 Diseases in fish 5 TBC

2 Fungi in aquatic plants 8 TBC

3 Field Trip Report (will be elaborated upon TBC TBC

confirmation)

Rules and Regulations of Aquatic Science Laboratory

1. ALWAYS use a lab coat and closed-toe shoes when in the laboratory.
2. NO eating, drinking or smoking in the laboratory.
3. While running an experiment, DO NOT use any electronic devices (e.g. mobile
phone, mp3 player, laptop … etc).
4. Your working space in the laboratory MUST be kept clean and tidy to avoid any
accidents.
5. When dealing with reagents/chemicals, READ the labels THOROUGHLY and ask the
laboratory technologist for clarification.
6. Take PRECAUTIONS when handling carcinogenic, mutagenic, toxic or corrosive
chemicals.
7. Please CONSULT the laboratory technologist before disposing any chemicals into
the sink.
8. In case of any EMERGENCY, get immediate help from the lecturer or laboratory
technologist.

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Practical Grading Rubric

Cognitive skills

Aspect Total marks Breakdown

(70m)
Introduction 8 6 – 8:
*provided in the Concise introduction with additional information (compared to what’s
manual provided in lab manual)

2 – 5:
Student provided additional information (compared to what’s provided
in lab manual)

1:
Copied from given lab manual

0:
None

Objectives 2 2:
*provided in the Copied from manual
manual
0:
None.

Material and 5 4 – 5:
Methods Copied and changed to passive voice
*provided in the
manual 1 - 3:
Incomplete copying; not changed to passive voice

0:
None.

Results 25 20 – 25:
Data tabulated/presented clearly; correct and complete interpretation
of results.

11 - 19:
Data tabulated/presented but results not interpreted correctly and
clearly.

4 – 10:
Data not tabulated nor presented clearly, results weakly interpreted.

1 – 3:
No raw data, results copied or weakly interpreted

0:
Wrong data/result interpretation.

Discussion 20 15 - 20:
Results are well-discussed; sound points given; able to relate back to
Objectives; honest interpretation of methodology; additional visual
data (pictures, graphs … etc) given
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 10 - 14:
Results are generally discussed, not much supporting points for results;
partially relates back to Objectives, few visual data given.

5 – 9:
Results not complete, no supporting points given, did not relate back
to Objectives, no or a few visual data given

0-4:
Weak or no discussion; no supporting points for results; did not relate
back to Objectives.

Questions 10 10:
Answered all question correctly

1 - 9:
Answered all questions but some incorrect

0:
Did not answer all questions; all questions answered wrongly.

Psychomotor skills

Aspect Total marks Breakdown

(30)
Conduct 10 5-10:
Handles equipment well, didn’t break any glassware or spoil any machinery, tidy
workspace.

0-4:
Rough with the equipment, break/spoil glassware or machinery, careless, messy
workspace.

Teamwork 10 5-10:
and Able to work in a team, able to help each other, able to delegate task amongst
Delegation selves.

0-4:
Unable/refuse to work in a team, withdrawn, unable to delegate.

Initiative 10 5-10:
Enthusiastic, present for all practical session, willing to volunteer or help, clean up
after practical session.

0-4:
No enthusiastic, absent for practical sessions, not willing to volunteer or help out
in lab, does not clean up after practical session.

** For field trips, marks for P and C skills will be graded according to rubric above, divided by 50 and 30, respectively and
multiplied with 10.

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 PRACTICAL 1

Diseases in Fish
Introduction
Fish is and has been a major source of food for the human population worldwide throughout recorded history as it
is rich in proteins and are readily available in all regions of the world especially areas nearest to the sea. Because
of its prominent role as a food source, fish are commercially farmed in mass aquaculture operations and sustaining
healthy fish in aquaculture operations is paramount in maintaining a steady supply of food for the population. All
fish carry pathogens, some of which cause diseases. In recent years, there have been more emergences of
diseases in aquaculture operations. Disease-causing pathogens include viruses, bacteria, fungi, mold and parasites
(metazoan, and unicellular).

For this course, we are going to look at several fish and examine the anatomy of the whole fish for diseases. We
will then culture the gut content for the fish dissected.

Objective
1. Detect physical signs of disease in fish
2. Learn to note and manually compile disease features in fish
3. Culture gut content of fish

Apparatus & Material

Section A
Stereo microscope
Compound microscope
Forceps
Scissors
Beakers
Petri dishes
Slides
Coverslips
Scalpel (or a dissection kit)
Saline solution

Methods

A. Observation of signs of disease:

1. Observe the outer body of fish for signs of disease.
2. As experiment goes along, observe all organs that are removed for signs of disease, according to Table
One.

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B. Dissection of fish -- Observation of diseased organs
1. Study the general characteristics of the external fish anatomy. Note down any abnormalities/changes
in color/fin condition/scale condition.
2. Illustrate and label the fish.
3. Remove the gills:
a. Cut through the bone at the top where the gills are joined to the head.
b. Cut through the bone at the bottom where the gills are joined to the head.
c. Lift the back edge (farthest from the mouth) of the gills and cut away from the skin (Each pair
of gills has 4 arches, each with a row of gill rakers. These rakes prevent food from entering the
gill and instead guide it into the throat)
d. Place gills into a petri dish with clean tap water. Put aside.
4. Opening the fish:
a. Cut the fish open beginning at the vent.
b. Do not cut too deeply or the internal organs will be damaged.
5. Internal anatomy:
a. Remove the eggs or milt by gently pulling the sacs away from the body.
b. Remove the liver and gall bladder by gently cutting any small membranes that join it to the
digestive system
c. Pull away from the stomach and remove.
i. Observe the digestive system by gently pushing a probe (8’’ spoon handle or
chopstick) through the mouth and into the stomach.
d. Remove the stomach by cutting it away at the throat and gently pulling.
e. Remove the complete digestive system and intestines (gut), which end at the vent. Put the
entire digestive system into a petri dish filled with sea/tap water. Put aside.
f. Remove the swim bladder by gently scraping it away from the sides of the body with the flat
side of the knife.
g. At the vent end of the fish, reach one finger under the swim bladder and pull it away. Put the
swim bladder into a petri dish filled with sea/tap water. Put aside.
h. Remove the kidney by cutting along each side. Use a spoon to lift it out.

C. Culture of gut content of dissected fish

1. Cut the gut or intestine laterally to open the intestinal or stomach tube into one flat piece.
2. Take a swab of the contents and spread it on the culture agar plate.
3. Incubate and observe weekly, for 2 weeks.

Results
A. Microscopic examination of parasites from the body, gills and gut of fish

1. Fill out Table 1 (below) after dissecting the fish.

2. Draw and label the anatomy of the fish.

Table 1: Organs and signs of diseases

Organs Symptoms Remarks Fish 1 Remarks Fish 2

Eyes Swollen, exopthalmia,
sunken, shape

Cornea Cloudy, deformed

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 Body Lesions, nodules, ulcers,
tumors

Scales Loss of scales, protrusion

Fins Intact, signs of rot,

nodules

Tail Intact, signs of rot,

lesions, nodules

Mouth Swollen tongue, lesions

cavity/tongue in mouth, presence of
ectoparasites
Gills Color of gills,
distribution/arrangement
of lamellae
Swim bladder Inflated, deflated

Liver Lesions, color

Stomach Filled with fluid, content

Results
1. Draw and label (as complete as possible) the anatomy of both fishes.
2. Complete Table 1.
3. Observe the culture of the gut content for both fish under the microscope.

Questions
1. What are the possible reasons for exophthalmia in a fish?
2. What are the two organs we look at first when purchasing fish from a market?
3. How would you know that a fish’s liver is in good health?
4. What is the ‘pyloric caeca’ and what is its function in the fish?
5. State the organ that determines how a fish swims.
6. State ONE common disease in tilapia.

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 PRACTICAL 2

Fungi in Aquatic plants

Introduction

There are a variety of pathogens which attack aquatic plants but the most common pathogen are fungi. Aquatic
plants such as seaweeds (marine) or water plants (freshwater) are prone to attacks by fungi because the spores of
these fungi exist naturally in water. Some examples of fungi diseases in aquatic organisms are as such:
Icthyophonus and saprolegniasis in fish; larval mycosis in shrimps and aspergillosis in corals. For this experiment,
you are going to detect possible fungal diseases attacking seaweeds and water plants.

Objectives

1. Learn how to isolate fungi from seaweeds and water plants.

2. Learn how to identify pathogenic fungi.

Apparatus and Material

Sterilized cotton swabs Bunsen burner

Fine scissors Parafilm
Forceps Clear Scotch tape
Gloves Slides
PDA agar plates Compound microscope
Inoculation loop

Methods

1. With gloves on, take out a few blades of the water plant from the aquarium.
2. Using a sterilized cotton swab, gently dab on any diseased area of the water plant blade.
3. Dab the cotton swab on the surface of the media on one NA (nutrient agar) plate.
4. Repeat steps 1-3 for two other blades from two different stems of the water plant [You should have three
different plates with swabs from three different parts of the plant].
5. Ensure all media plates are placed upside down.
6. Seal the plates with parafilm so that the lid cannot be misplaced.
7. Place the agar plates into an oven at 20-22°C for 24 hours.
8. After 24 hours, check if there are any signs of colonies growing.
9. Repeat Steps 7 and 8 for five days, noting down any new colonies and the characteristics of fungal
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 growth. Close the petri dish after each time of inspection.
10. Cut some clear tape, sticking one end of your thumb and the other end to your middle finger.
11. Uncover Petri dish and prepare to take a sample.
12. Using your index finger of the same hand with the sticky tape, press down your index finger onto the
media surface to sample the fungal growth.
13. Gently press the media to sample as much fungi as possible and then lift the tape slowly off the media
surface.
14. Place the tape onto the slide
15. View the slide under a compound microscope.
16. Note/draw what you see.
17. Repeat steps 12-16 for each media plate.

Repeat the steps above for seaweeds.

Results

1. Draw what you have observed in your respective slides.

2. Characterize the fungi/mold you have drawn according to the charateristics and descriptions given below.

Characteristic Points to note

Spores Color, shape, features, size, arrangement (basidia, asci etc…)
Hyphae Septate/aseptate, branching or not, pigmented or not, even or uneven in width,
composed of arthroconidia or pseudohyphae

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Questions

1. State THREE differences between seaweeds and water plants.

2. Why should you use nutrient agar (NA) instead of blood agar when culturing fungi?
3. State THREE common fungal diseases of aquatic plants.
4. What role do fungi play in the marine ecosystem?

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 PRACTICAL 3 TBC

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