AQUATIC ANIMAL RESPIRATION

By :
Name : Mellya Rizki Pitriani
Student ID : B1B017031
Group : VI
Subgroup :2
Assistant : Ita Purwati

PRACTICAL REPORT OF ANIMAL PHYSIOLOGY II

MINISTRY OF RESEARCH, TECHNOLOGY AND HIGHER EDUCATION

JENDERAL SOEDIRMAN UNIVERSITY
FACULTY OF BIOLOGY
PURWOKERTO
2019
 I. INTRODUCTION
A. Background

Respiration is the process of taking O2 from the environment into the

animal's body and releasing CO2 into the environment. Respiration in aquatic
animals (fish) includes taking O2 from the waters. O2 consumption per unit time
shows the metabolic rate in fish. The intensity of fish breathing decreased with
increasing fish weight. The rate of consumption of O2 also decreases with the
availability of small amounts of O2 for fish. The O2 concentration threshold growth
will be higher at high temperatures, coinciding with higher O2 consumption rates,
changes that are equal to the threshold for increasing activity or feeding rates in fish.
Fish activities, including developing foods that will be converted into fish meat, will
be limited by the concentration of O2 that is common in water (Yuwono, 2001).
The exchange of oxygen and carbon dioxide between an organism and its
environment is known as aerobic respiration. Anaerobic respiration is principally
carbon dioxide given from certain organisms with no oxygen taken. Oxygen needs
are obtained from the composition of carbohydrates and fats in the body (Weichert,
1959).
The source of dissolved oxygen can come from the diffusion of oxygen in the
atmosphere (about 35%) and photosynthetic activity from aquatic plants and
phytoplankton. The diffusion of oxygen from the atmosphere into water can occur
directly under stagnant water conditions. Diffusion can also occur due to agitation or
upheaval of water masses due to waves or waves and waterfalls. But in essence, the
diffusion of oxygen from the atmosphere to the waters takes place relatively slowly,
despite the turbulence of the water mass. Therefore, the main source of oxygen in the
waters is photosynthesis (Effendi, 2003).

B. Purpose
The purpose of this laboratory activity is to calculate the rate oxygen
consumption in aquatic animal.
 II. MATERIAL AND METHODS

A. Material
The materials that used in this practice are freshwater fish, nila fish
(Oreochromis niloticus), nilem fish (Osteochillus vittatus), KOH-KI, H2SO4,
Na2S2O3, MnSO4, and amylum reagent.
The tools that used in this practice are aerator, technical scales, respirometer,
erlenmeyer, Winkler bottle, burette, statif, and measuring cup.

B. Methods
The methods that used in this activity are:
1. Respirator is being run, the water pump is being turned on for 15 minutes.
2. The weight and volume of fish are measured (Vfish = Vtotal – V1(initial))
3. The fish is put into the respirometer, no bubbles inside are made sure.
4. The water sample is taken with Winkler bottle (250 mL).
5. The solutions are added
a) MnSO4 1 mL
b) KOH-KI 1 mL
c) H2SO4 1 mL
6. The solution is taken into the erlenmeyer (100 mL), 2-3 drops of amylum is
added.
7. The early titration with Na2S2O3 is done.
8. After 15 minutes, the water sample is taken with Winkler bottle (250 mL).
9. The final titration is done.
10.The consumption of O2 is calculated.
1000
Ota/Otak = ×p×q×8
100
VO2= (cO2i - cO2f) × V × H-1 × W-1
 III. RESULT AND DISCUSSION

A. Result
Tabel 3.1 The Result of Consumption of O2 in Entourage VI
VO2
H Volume cO2i cO2f
No. Fish Weight (g) (mg/L/h
(Hour) (mL) (mg/L) (mg/L)
our)

1. Large Nila 0,25 130 124 5,6 5,6 0

2. Small Nila 0,25 20 20 5,4 6 -0.6543

3. Large Nila 0,25 9055 131 7,4 7 0,110

4. Small Nila 0,25 5435 26 6,4 4,8 1,337

Calculation of animal respiration :

1000 1000
- Ota = ×p×q×8= × 2,7 × 0,025 × 8 = 5,4 mg/L
100 100
1000 1000
- Otak = ×p×q×8= × 3 × 0,025 × 8 = 6 mg/L
100 100
- V = Vrespirometer – Vfish
= 5465 - 20
= 5445 mL
= 5,445 L
- vO2 = [ (Ota - Otak) x V ] / H x W
= [ (5,4 - 6) x 5,445 ] / 0,25 x 20
= -0,6534 mg/L/h
B. Discussion
Based on the data that group 2 got, the sample that used is small nila with
5,445 L volume, 20 g weight, early oxygen dissolved is 5,4, late oxygen dissolved is
6 and oxygen consume is -0,6534 mg/L/h.
According to Ross (1999), the respirometer works on a principle that in breathing
there is oxygen used by the organism and there is carbon dioxide released by it. If the
breathing organism is stored in a closed space and carbon dioxide released by an
organism in a closed space is bound, air depreciation will occur. Based on the
statement, the oxygen dissolved after the treatment is should be decreased, but in our
data is late oxygen dissolved is higher than early oxygen dissolved. So, it might be
there is a something wrong, we expect that in the reading the scale during titration
was not correct.
The relationship of consumption of O2 with the metabolic rate is the
consumption of O2 at the maintenance metabolic rate is less than 60% higher than
fish during feed shortages. The consumption of O2 in growing fish feeds originates
from one side, from maintenance metabolism and from the other from the synthesis
and the rate of consumption of O2 decreases with a decrease in the availability of
oxygen for fish. The relationship between KO2 and metabolism is that the
metabolism requires oxygen, the more or the faster the metabolic rate the higher the
O2 consumption needs. So that the more KO2, the more need for hemoglobin which
functions to bind oxygen in the blood (Zonneveld et al., 1991).
Dissolved oxygen is needed for respiration and metabolism and the survival
of organisms (Mulyani, 2014). Fish are aquatic animals that consume oxygen
dissolved in water. Taking oxygen in fish is carried out by the main respiration
organ, the gills. The amount of oxygen uptake through gills can be measured by the
method of static water or running water. The oxygen consumption parameter is used
to assess the rate of fish metabolism, because most fish energy sources come from
aerobic metabolism. Fish need to consume oxygen to do aerobic metabolism,
therefore changes in oxygen consumption of fish can be used to assess changes in
metabolic rate. The metabolism of poichilothermic animals is affected by changes in
ambient temperature. When temperatures are low, metabolism decreases and
metabolism will increase at increased environmental temperatures (Sudibyo, 1999).
Three fundamental metabolic variables are standard metabolic rate (SMR),
maximum metabolic rate (MMR) and the scope of aerobic metabolism (AMS). SMR
is defined as minimal maintenance or resting metabolic rate from non-depressed,
post-absorption and non-proliferating ectotherm adapted to experimental conditions,
below this physiological interrupted function. Therefore SMR is a basic cost of living
and is very functionally important. For example, the theory of life history argues that
the energy obtained is allocated between functions such as growth, reproduction and
self-maintenance. At the other end of the metabolic scale, the MMR provides the
upper limit for aerobic energy metabolism. MMR is conventionally measured at the
point of fatigue. A similar term is the active metabolic rate (AMR), i.e. measured
using swimming respirometry and reflects the metabolic rate at maximum continuous
swimming speed. Reducing SMR from MMR provides an AMS measure, which
represents the total amount of aerobic energy available to animals for processes that
require energy including digestion, locomotion, growth and reproduction (Rosewarne
et al., 2016).
The total oxygen consumption of fish reflects basal metabolic status and is
one indicator of the general health of fish. It may also be useful to assess the
physiological state of an organism, assist in evaluating susceptibility or potential
resistance, and is also useful for linking animal behavior which ultimately becomes a
predictor of functional disorders of the population. Fish are heterotrophic animals
because they are often used as bioindicators of stress pollution associated with
biological early warning systems. Therefore, the consumption of differential oxygen
can be used as a bioindicator of stress related stresses in biological early warning
systems (Neelima, 2016).
Metabolic rate is the total amount of energy produced and used by the body
per unit time. The metabolic rate is closely related with respiration because
respiration is the process of extracting energy from molecules food that depends on
the presence of oxygen. Metabolic rate usually estimated by measuring the amount of
oxygen consumed living things per time unit. This allows due to oxidation from food
ingredients require oxygen (in known quantities) for produce known amounts of
energy too. However, the pace metabolism is usually sufficiently expressed in terms
of the rate of oxygen consumption (Putra, 2015).
In this practical activity, we used nila fish (Oreochromis niloticus) and nilem
fish (Osteochillus vittatus) for other entourages. There must be reasons why those
two kind of fish are used. It is because nilem fish and nila fish can be found easily in
our area, Indonesia for specially because those fish are common and popular to
consume by people and also cultivated intesively (Prakoso and Chang, 2018).
The steps of the titration method by means of Winkler are as follows (Alaerts,
1987):
1. The sample water was put into a 125 mL Winkler bottle (at the time of
practicum, one bottle of water was used, about 100 mL) with the condition
that there was no air entering the sample.
2. Water in a Winkler bottle plus MnSO4 solution as much as 0.5 mL (at
practicum time, the function of MnSO4 solution is binding oxygen, MnSO4
solution used as much as 1 mL or 21 drops), and KOH-KI solution as much
as 0.5 mL (at practicum time, the function of the KOH-KI solution is to bind
gases other than oxygen, the KOH-KI solution used is 1 mL or 21 drops). The
solution is homogenized and then left so that a heterogeneous layer is formed,
which is in the upper part of the clear and in the bottom is a brown precipitate
(if it does not contain oxygen, white colored deposits).
3. The water in the Winkler bottle is reacted with H2SO4 as much as 0.5 mL (at
the time of practicum, the function of H2SO4 solution is to balance or
neutralize a substance, KOH-KI solution is used as much as 1 mL or 21
drops), then shaken so that the sediment becomes dissolved and formed
yellowish liquid and left for 10 minutes.
4. The water in the bottle is taken 100 mL stored in the erlenmeyer tube and
added starch, as a color indicator of 11 drops and then titrated with Na2S2O3
0.025 N so that the yellow color from the initial mixture becomes clear.
5. The Winkler method is done twice to get the average value.
6. Soluble O2 formula:
Ota = 1000/100 × p × q × 8
Information:
Ota: initial dissolved oxygen (mg / L)
p: used Na2S2O3 solution
q: normality of Na2S2O3 (0.025)
8: molecular weight of oxygen
Measurement of fish oxygen consumption is done by placing test fish in
running water. The day before measuring oxygen consumption, the fish was fasted
for 24 hours. Measurement of dissolved oxygen concentration was carried out by
Winkler titration. Oxygen consumption (mg / j / g) is calculated by formula
(Goenarso et al., 2003):
VO2 = (cO2i-cO2f) × V / H × W
Information:
VO2: oxygen consumption (mg / g / hour)
cO2i: initial dissolved oxygen (mg / L)
cO2f: final dissolved oxygen (mg / L)
W: fish weight (g)
V: tube volume after reduced fish volume (L)
H: interval of oxygen measurement beginning and end (hour)
The function of the solution used for the titration process includes the
following (Wetzel & Linkens, 2000):
1. MnSO4 and KOH-KI: to form brown deposits, indicating that there is still O2 in
the sample. If the resulting deposits are white, there is no O2 dissolved in the sample.
KOH itself functions to reduce MnSO4.
2. H2SO4: converts a solution that is initially cloudy brown to clear brown, and to
break or remove bonds that occur due to the influence of a solution of KOH-KI,
MnSO4. This solution is not formed from the reaction between sulfuric acid and
manganese oxide to form manganese sulphate.
3. Amylum: to detect the presence of starch in solution and as an indicator that
changes the color of the solution that was originally clear brown to light blue.
4. Na2S2O3: for titration as a p value to find dissolved O2 levels.
The factors that influence the level of oxygen consumption are divided into
two, namely the external and internal factors. External factors are influenced by the
partial pressure of oxygen and temperature. Increased metabolic rate will follow an
increase in temperature to a certain extent. The internal factor is that which is directly
related to the fish itself, such as the size of fish, activities, conditions of fish health,
and sex. Other factors that influence oxygen consumption in fish include:
1. Activities, fish with high activity such as fish that actively swim will consume far
more oxygen than fish that are not active.
2. Size, the metabolic rate of small fish is higher than large fish so that more oxygen
is consumed.
3. Age, young fish will consume more oxygen than older fish.
4. Temperature, fish that are at high temperatures also have a high metabolic rate so
that more oxygen is consumed.
5. Gender, females do more respiration because females have a more complex
hormonal system than males ..
6. Emotions / stress, the higher the emotion, the more respiration is done because of
certain hormones that affect metabolism so that respiration is faster (Zonneveld et
al., 1991).
 IV. CONCLUSION
Based on the result and discussion, it can be concluded that early oxygen
dissolved is 5,4, late oxygen dissolved is 6 and oxygen consume is -0,6534 mg/L/h in
small nila fish.
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