IDENTIFICATION OF ZOOPLANKTON ORGANISMS IN THE AQUACULTURE PONDS –

INSTITUTE OF FISHERIES OF MARINE SCIENCES

A special project and General fundamental (ZOO1)

Ashley Abasula
Mark Joshua Bunom
Virgilio Bataller
Alona Jen Quibral
Laurence Quintana

2025
 INTRODUCTION
Zooplankton are small, often microscopic creatures that float in the water of aquaculture ponds. They are

important to the ecosystem because they provide food for fish and other water-dwelling animals. Zooplankton,

which includes tiny invertebrates like crustaceans, rotifers, and protozoa, are essential for keeping aquaculture

systems balanced (Azim et al., 2022). They are an important food source for fish and other aquatic creatures,

and they help break down organic matter and recycle nutrients in the ecosystem (Huang et al., 2023)

Aquaculture is an important part of the world’s food industry, offering a major source of protein for

people (FAO, 2020). Aquaculture is a rapidly growing industry that contributes significantly to global food

security, particularly by supplying high-protein seafood. Proper identification and monitoring of zooplankton

organisms can provide valuable insights into water quality, feed dynamics, and overall pond health

(Madhusudhan et al., 2021). Among these, zooplankton plays an essential role as a primary or secondary food

source for aquaculture species, such as fish larvae and juvenile stages. These small invertebrates, which include

crustaceans, rotifers, and protozoa, are key foods for fish and other aquatic creatures. They also help with

nutrient cycling and impact water quality (Huang et al., 2013).

Although zooplankton communities are very important, there is limited information about their

composition and diversity in aquaculture systems in the Philippines, particularly in the CNSC-IFMS

aquaculture pond. This gap in knowledge makes it hard to develop effective management strategies that could

improve aquaculture productivity and maintain a balanced ecosystem. (Li et al., 2023).

This study will help create sustainable aquaculture practices, ensuring that aquaculture operations can

thrive in the long run while also protecting aquatic ecosystems. The results of this study will offer important

information about the ecology of zooplankton in tropical aquaculture systems, guiding the creation of effective

management strategies to boost aquaculture productivity and keep ecosystems balanced.

 The primary objectives of this study is to determine the different kinds of zooplankton found in our

campus pond. More specifically, this research intends to. 1) Isolate zooplanktons using towing methods.

2)Identify the isolated zooplanktons based on morphology characteristics. 3)Determine the species biomass of

the zooplanktons.

This research aims to identify the zooplankton species found in the ponds at CNSC-IFMS and to

explore the types of zooplankton using a towing method. The study will take place from March 2025 to May

2025 at the Biology Laboratory in the Mercedes Campus of Camarines Norte State College.

Author’s and Title Type Objective Method Conclusion

Year
Ekelemu and Study on the Rol- Phytoplankton Approximately The identified
Nwabueze Zooplankton International signify the 10 liters of zooplankton were
(2020) Production in biological riches water were categorized into 4
Ponds of aquatic randomly orders: copepoda,
Under Different environments and collected from rotifera,
Fish Farming . Zooplankton, chosen sites, cladocera, and
System in West source for combined, and Diaptomus. The
Bengal omnivorous fish filtered. The primary
in aquaculture, obtained zooplankton
serve an plankton was groups present in
important concentrated to all the samples
function in open 20 ml and were noted to
water ecosystems. stored in 4% Copepoda and
Their absence formalin for Cladocera are
may adversely examination. represented by
affect the survival Cyclops sp. and
of fish spawn in Daphnia sp.,
nursery ponds. respectively
Zooplankton are
typically divided
into Rotifers,
Cladocerans, and
Copepods
Diversity and Rol- Zooplankton rely Three Zooplankton play
Bai, X.; Jiang, Community international on a variety of replicated a crucial role in
Y.; Jiang, Z.; Structure of factors, including samples were the aquatic,
Zhu, L.; Feng, Zooplankton in climate change, gathered for the greatly enhancing
J (2022) Homestead types of habitats, ecological aquatic biological
Ponds of a physico-chemical parameters and productivity
Tropical Coastal characteristics, zooplankton
Area and biotic communities of
 influences. the homestead
Environmental ponds, and their
variables average values
significantly were calculated
influence the
distribution of
zooplankton
Pearson, A. A., A global review Rol- water-dwelling employing a zooplankton
& Duggan, I. of zooplankton international creatures, referred horizontal communities in
C. (2020). species in to as aquaculture, launch freshwater
freshwater entails the net featuring a aquaculture
aquaculture creation and 40 μm mesh. ponds around the
ponds: maintenance of Samples were world to assess;
what are the aquatic maintained 1) the occurrence
risks for organisms mainly using of
invasion? intended for Ethanol was non-native
human sent back to the species, 2) the
consumption. laboratory for standards of the
Water-based identification. research
organisms are Zooplankton conducted, 3) the
frequently were extent of
cultivated for recognized representation of
purposes other through a key freshwater
than food. compound aquaculture
purposes, microscope. countries in
including for the (Olympus BH- researches, and 4)
decorative market 2) and/or a the depiction of
or to dissecting leading
act as natural pest microscope aquaculture
control agents (Olympus species
SZ60) as
needed
(Tumwesigye Zooplankton Rol- local zooplankton Plankton Paoay Lake in
et al. 2022 Composition species identified sampling was Luzon Island,
(DA- and Diversity in Paoay Lake performed to An investigation
BFAR 2022). in Paoay Lake, and to offer gather the in the Philippines
Luzon Is., essential data samples. was carried out to
Philippines regarding species employing a refresh the earlier
composition and conical lists of
variety. It also plankton net zooplankton
seeks to offer (Diameter: species in the
updated 0.254m; Mesh region and
information on size: 80μm) species that have
certain Two samples, were previously
physical and each with 3 noted as rare in
chemical factors replicates, were earlier studies but
such as depth gathered from were often found
various zones at all sampling
within each locations
sampling
 location that
were collected
throughout the
day (roughly
from 0800 to
1000 hours)
from each of
the four
sampling
locations in the
lake

Pinti (Ilac et Zooplankton Rol- local Zooplankton are Exterior marine

al., 2024; Morphology, essential. Samples were zooplankton in
Mendoza et al., Abundance, and highlighting their gathered the Ilocos Sur
2023)et al. Their environmental employing a Area
(2023) Relationship significance. handheld fine This research
with They play a role mesh plankton carried out in
Physicochemical in ecological net (50 μm) and Brgy. was driven
Parameters in processes like a 10L container. by motivation.
Coastal Waters carbon capture Water was Fuerte, Caoayan,
of Fuerte, via the biological passed through Ilocos Sur, to
Caoayan, carbon pump, a the net, cleaned, create fresh
Ilocos Sur process that and moved into perspectives on
supports in 100ml empty the
moving carbon to containers. zooplankton
deeper layers of Samples were population. It
the oceanl marked with the seeks to examine
date and time zooplankton
when they were structure,
collected quantity, and
physicochemical
Bakhtiyar Y, The Importance ROS- Most importantly, Testing This chapter
Arafat MY, of Plankton in international algae are the sole different dietary examines the
Andrabi S, Marine Fish and source of these combinations, nutritional and
Tak, HI (2020) Shellfish Larval linoleic and experimental biochemical
Nutrition linolenic acids. microdiets makeup of
Consequently, provide an plankton,
zooplankton opportunity to emphasizing their
either requires explore substantial levels
essential fatty potential of proteins,
acids from algae macronutrient lipids, vitamins,
directly or preference minerals, which
transforms them are vital for
from linolenic marine larval
acid stage
Minaz and Relation among Ros- Aquaculture Using filtered In aquaculture,
Kubilay, 2021 zootechnical international activities can water from the zooplankton are
Raza et al., performance, offer a valuable River Nile, frequently
2024 biochemical supply of sugarcane cultivated and
indicators, water biological molasses was utilized as live
quality, and substances diluted with the food to promote
small essential for water in a the development
invertebrates aquatic creatures, beaker prior to of fish larvae
(zooplankton) including being
abundance proteins,etc introduced into
reared in numerous other the fermentor
biofloc- bioactive
supplemented materials
system
International The effect of Ros- A key benefit of Three The current
Journal of nursery international this system is the production research aims to
Zoological production access to natural systems were evaluate the
Investigations system (in cage food for the fish, assessed for the growth
Thirumala, and pond) on which nursery stage of performance,
Harish performance, consequently aids tambaqui: health status,
KumarK, BR health status, in lowering earthen ponds natural food
Kiran Boyd et and plankton production (P), cages in consumption,
al.,( 2020) ingestion of the expenses ponds (CP), and muscle fiber
low trophic level cages in a characteristics,
fish tambaqui, reservoir ©, and isotopic
Colossoma with four makeup of
macropomum repetitions for tambaqui raised
each for 56 days in
various
production
systems
Marilao and Plankton Ros- local Microscopic The sampling Phytoplankton
Meycauayan, Community plankton act as a was conducted constitute
Bulacan, Structure of the main food source in three main producers in
Philippine Aquaculture for fish and can repetitions for an aquatic
Pleto, J. V. R., pond indicate levels of each. environment that
& Cabillon, Y. pollution. lake. The can influence
C. L. (2022). The research gathered the efficiency of
concentrated on plankton the system. They
the makeup of the samples were hold a significant
phytoplankton put in 50-mL role
and zooplankton bottles with function as main
populations in screw caps, producers and
fish pools along promptly influence upper
with the MMROS placed in 10% trophic level
formalin
solution, and
conveyed to the
laboratory for
examination
and
recognization
 Cavite, Ysabel Species Ros- local The classification Every sample Research on
& Juan, Janna composition of of freshwater of zooplankton freshwater
& KOH, freshwater zooplankton from from each zooplankton in
MOSES, & zooplankton groundwater is chosen the Philippines is
Lopez, Louie fauna from still not well sampling primarily
& Papa Rey selected understood, but locations were confined to
donne. (2020) groundwater- understanding replicated in surface bodies of
dependent this group is triplicate using water like rivers
ecosystem in essential to know conical and lakes
Bulacan plankton nets and entirely
province featuring mesh ignoring
(Philippines) sizes of 45 µm groundwater and
and 60 µm groundwater
reliant
ecosystems such
as caves,
accessible wells,
springs and
groundwater
pumps with
piping

The zooplankton will be collected in CNSC- IFMS Aquaculture ponds In order to harvest zooplankton, water
must first be filtered using a net, then collected using bottles, or water samplers. The choice of appropriate
equipment, including the mesh size of the netting material, the time of collection, the water depth of the study
area, and the sample technique, will be crucial to the effectiveness of the sampling
Using a plankton net is the most popular way to capture zooplankton. More water has been filtered, and
the equipment can be used for both qualitative and quantitative research. Different sizes and types of plankton
nets are employed. The various nets can be roughly divided into two groups: closed nets with messengers for
gathering vertical samples from desired depths, and open nets used primarily for oblique and horizontal hauls.
Deployment: Towing Method
To prevent sedimentation, pull the plankton net obliquely until the bottle (at the end of the net) is filled
with water from roughly 0.5 meters above the bottom to the surface. Plankton net with a mesh size of 40 µm.
20 cm in diameter, and an outlet tap can be use in sampling.
The optimal vertical-horizontal haul is one that is completed quite quickly and consistently (around one
meter every two seconds). The use of Horizontal Tow: To sample a particular depth or area, the net is towed
horizontally. And a Vertical Tow: To sample the water column, the net is lowered to a certain depth and then
raised. Start to tow after pulling until the net is expanded. Bring the net inside after a few minutes and leave it
hanging for a while.
Collection and Retrieval
Zooplankton are filtered out of the water when the net is towed, and they gather in the bottle. Running
ambient (fresh) water from the net’s exterior will wash all of the plankton into the bottle. After that, samples are
taken from the cod end and put into 500mL–1L sample jars or a clean, dry glass beaker. It is necessary to
remove any unnecessary material or debris. The following collection, the sample is stored for subsequent
analysis using an appropriate fixative.
Fixation and Preservation of samples
Fixation The following can be used to fix samples: For morphological investigations, buffered formalin
(4–5%) is frequently used as a fixative and preservative. Ethanol 95–100% non-denatured for molecular
research. Lugol’s iodine solution for examination under a microscope For additional short-term storage and
preservation, 25% vinegar (3% acetic acid solution), 30% ethanol, or 30% glutaraldehyde might be utilized.
Crucial Points to Remember:
Preservation of pH: Over time, formalin solutions may turn acidic, which may cause some zooplankton
species’ calcareous structures to dissolve. To assist maintain calcareous structures, borax can be added to
formalin to bring the pH up to about 8. Long-term preservation: Make sure samples are kept in sealed
containers with enough preservative for long-term preservation. Sample handling: Be cautious not to lose any
zooplankton during sample transfers.
Labelling: Because regular ballpoint pens disintegrate in preservative solutions, use archival-quality inks
and pens for labelling. Storage Requirements: Keep preserved samples below 25°C in a well-ventilated room.
Techniques for fixing formaldehyde: There are two ways to maintain and repair zooplankton: The first approach
Fill a 500 ml empty container with 300 ml of seawater and 50 mL of neutralized 40% formaldehyde. After
adding the concentrated, drained, or living zooplankton samples, seawater is poured to the container. To mix
the contents, temporarily screen down the jar and invert it a few times.
First, half the amount of seawater or distilled water containing the 4% neutralized formaldehyde is added.
A jar that is empty Then, using the least amount of seawater possible, the recently extracted and concentrated
samples are introduced. Then, until the jar is filled, more of the 4% formaldehyde solution is poured. After a
day or two, the fixed samples are filtered once again. In the end, the animals are kept in either 75% ethanol or
the neutralized 5% formaldehyde solution in tiny jars with plastic screw covers.
Samples are preserved. Carefully decant the fixative and replace it with the preservative. Add the
appropriate amount of concentrated preservative (2.5 ml of buffered 40% formaldehyde or 7.5 ml of
preservative with additives per 100 ml container volume) to the sample jars, filling them roughly three-quarters
of the way with seawater or distilled water. After that, fill the jar with seawater or purified water and cover it
tightly. To disperse the animals throughout the preservative, it is preferable to invert the jar multiple times.
Sample bottles should ideally be kept in a cold, dark place and should be examined every month or every three
months for evaporation or animal condition. Wide-mouthed glass bottles with screw-on plastic covers are the
best sample storage vessels. High-quality polypropylene-lidded polyethylene jars. The plankton storage bottles
should be properly labelled.
Labels may include the following details: Number of the station The sampling date Time spent
sampling. Depth of sampling. Net type, mesh size, and mouth size. Haul type (vertical, and horizontal, ) The
number of tums that the flowmeter recorded Name of the collector Information on the outside label No bottle.
Station number The sampling date Day and Night The sky Duration depth of the sampling location Net type
Mesh opening Reading from a flowmeter Name of the collector
 In order to prevent bacterial action and autolysis from damaging animal tissue, the samples should be
stored with buffered formalin as soon as possible after collection, preferably within five minutes. The perfect
fixative should be inexpensive and has a rapid rate of animal death. Once more, it must not be poisonous or
corrosive. The most common uses for buffered formalin (4-5%) are as a fixative and a preservative. 70%
ethanol or 40% isopropanol are the additional preservatives that are utilized. Other fixatives that are
occasionally employed include acetic acid, picric acid, ethanol, and others. Formalin. To prevent unwanted
osmotic effects, the concentrated formalin should be diluted with fresh water, seawater, or ideally, water from
the sampling region. One part formalin to nine parts fresh water or seawater is the dilution ratio. The fixative
should have a pH of about 8.0. Using buffered formalin is advised. FAA stands for formalin-acetic acid. It is
made by mixing equal amounts of glacial acetic acid (an alterative) and formaldehyde (37%). A 250 mL sample
is mixed with 5 ml of FAA. With the exception of “naked” flagellates, the majority of species do well in FAA.
Examination of the Data
Biomass.
The amount of living matter or live weight in the zooplankton sample is referred to as biomass. Fishery
potential and secondary productivity in the research region are assessed using the value that was acquired. The
methods listed below are used to estimate the biomass. The displacement volume method is used to calculate
the total zooplankton volume. Use a piece of dry, clean netting material to filter the samples; its mesh size
should be the same as or less than the net’s mesh size. The filter paper is used to remove the water that exists
between the organisms. Using a spatula, move the filtered zooplankton to a measuring cylinder filled with 4%
buffered formalin. Determine the displacement volume, note how much fixative is displaced in the measuring
jar by The zooplankton. By bringing the sample to a known volume in the measuring jar, you may determine
the settled volume. Before recording the settled volume, let the plankton settle for at least twenty-four hours.
Getting ready for light microscopy.
Mounting and staining.
While raw material in a water mount can be used to swiftly inspect and identify some planktons, there are
restrictions. To perform a more thorough analysis, place a tiny drop of the plankton sample on a sterile
microscope slide. After letting the water evaporate, heat the slide until it is completely dry. To the plankton
sample on the slide, add a drop of the diluted methylene blue solution. Depending on the plankton’s density and
the desired staining intensity, let the stain sit for one to three minutes. If more stain is needed, gently rinse the
slide with pure water. To get rid of extra water, blot the slide using absorbent paper. Add glycerin to help keep
the colors of the zooplanktons and to stop specimens from shrinking and the material from drying out. Put a
glass slide over it and look through the microscope.
Permanent slide preparation
Fixing Samples: Dehydrate samples by passing them through various alcohol grades: 30% and 50% for 30
minutes, and 70% for a few days. Wash specimens with distilled water. Staining: 30 seconds to 1 minute is the
staining period. The specimen needs to be cleaned with distilled water after submersion. By submerging the
specimen in an acid-alcohol mixture (70 percent ethanol and a few drops of HCI), excess stain can be
eliminated. Mounting After the stained specimen has been held in glycerin or clove oil overnight to produce
transparency and lessen the effect of the alcohol treatment, it is cleaned with a succession of dehydrating baths
(70%, 50%, and 30% ethanol).Using a glass rod, apply a drop of mountant to the slide. Move the stained
sample to the drop. To prevent air bubbles from being trapped in the mountant, place the cover slip at one side
of the drop and gently lower it. Apply nail polish or wax to seal, then properly label the slide. Do not use the
slide until it has dried, and keep it horizontal. Heating in an oven at roughly 35 °C might shorten the drying
time.
Understanding the distributional pattern.
Seasonal variability, and community structure of zooplankton in an aquatic ecosystem requires species
identification. It is a specialized task that calls for perseverance, expertise, and a substantial body of published
work. With the use of illustrated checklists, common species might be initially identified. The specimens that
have been identified and labeled ought to be appropriately stored for future use. Here are some examples of
zooplankton identification guides.
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