AQUATIC ANIMAL RESPIRATION

By :
Name : Pratiwi Kusuma Kurniawati
Student ID : B1B017007
Entourage : VI
Group :3
Assistant : Afif Ghalib Ammar Zain

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

Oxygen is a compound needed by almost all living things on this earth. In

the study of physiology oxygen is used in metabolic processes, namely fuel for
oxidizing food substances. Only a few animals can fulfill their energy without
oxygen, that is by utilizing the chemical energy of organic compounds
anaerobically but only produce enormous amounts of energy (Suharsono et al.,
2018). Oxygen (O2) in its molecular state is essential for many metabolic processes
that are vital to aerobic life. Like all aerobic organisms, fish are susceptible to the
effects of reactive oxygen and have inherent and effective of different biotic and
abiotic factors on antioxidant defenses in fish (Neelima et al., 2016).
The survival of fish is largely determined by ability fish in using oxygen
from their environment dissolved oxygen in the waters, will affect the physiology
of fish respiration, and only fish that have a respiratory system that is suitable and
able to adapt to the environment that can survive (Malini & Muliani, 2016). Total
oxygen consumption of fish reflects its basal metabolic status and is one of the
indicators of the general health or/and well being of the fish. It may also be useful
to assess the physiological state of an organism, helps in evaluating the
susceptibility or resistance potentiality and also useful to correlate the behaviour
of the animal, which ultimately serve as predictors of functional disruptions of
population (Neelima et al., 2016).
Oxygen consumption is used as an indicator of metabolism in fish, and
differences in salinity affect the energy needed for osmoregulation in some species.
Respiratory response is different from the difference in salinity among teleos
species. Low oxygen consumption figures are obtained in isosmosis salinity. Low
oxygen consumption in fresh water and its use increases with increasing salinity.
This shows that fish in seawater have lower oxygen consumption than freshwater
(Seeley et al., 2003)
Respiration is the process of taking oxygen from the environment into the
animal's body and removing carbon dioxide from the body into the environment.
Respiration in aquatic animals, examples of fish include extraction of oxygen from
water. The metabolic rate in fish is indicated by consumption of oxygen per unit
of time. Fish intensity from fish respiration decreases with increasing fish weight.
 The rate of consumption of oxygen also decreases with the availability of small
amounts of oxygen for fish. The oxygen concentration threshold growth will be
higher at high temperatures, coinciding with higher oxygen consumption rates.
Changes are the same as the threshold for increasing activity or feeding rate in fish.
Common oxygen concentrations in water will limit fish activity, including
developing foods that will be converted into fish meat (Yuwono, 2001).

B. Purpose

The objectives of this laboratory activity are:

1. To measure oxygen consumption of aquatic organisms either by titration
(Winkler method).
2. To measure metabolic response of aquatic animals related with the body weight
as well as environmental changes or stress.
 II. MATERIALS AND METHODS

A. Materials

The materials that used in this practice are nile tilapia (Oreochromis
niloticus), nilem (Osteochillus vittatus), reagent for titration the oxygen content
such as KOH-KI solution, a solution of concentrated H2SO4, MnSO4 solution,
Na2S3O3 solution, and amylum reagent.
The tools that used in this practice are aerator, technical scales, meausring
cup are large, gauges of oxygen consumption (respirometer), thermometer,
Winkler bottles, erlenmeyer, burette, and stative.

B. Methods
The method used in this practical are :
1. Respirometer to be used in the experiment is prepared.
2. Body weight of aquatic animal is weighed with an analytical balance.
3. The volume of aquatic animal is measured by using a large measuring cup.
Difference in water level before and after the fish entered is measured.
4. The fish is inserted into the respirometer (in tube I), so no air trapped in it and
let the test animal a few minute in it in order to aclimited.
5. First sampling of water (early) was performed using Winkler bottles (volume
125 ml), the water out through tube I hose from the tube I.
6. The airator is switched off after the fish is inserted and water hose of tube I is
closed and left for approximately 30 minute.
7. Measurement of dissolved oxygen in water samples I is using by titration
methods.
8. A total of 1 mL aqueous KOH-KI is added to the Winkler bottle containing the
water sample and shaken until homogenous.
9. A total of 1 mL solution of MnSO4 is added later seen whether there are
sediment or not.
10. A total of 1 mL of concentrated H2SO4 is added.
11. The sample solution was transferred into a measuring cup of 100 mL water
sample so that the color looks golden yellow or light brown.
12. The sample solution was titrated by adding a water solution of Na2S3O3 so the
previous color, light blue became clear (no color).
13. The value of CO2i / ota (initial dissolved oxygen) is calculated.
1000
Ota/cO2i= xpxqx8
100

Description:
Ota: initial dissolved oxygen (mg / L)
p: used Na2S2O3 solution
q: normality of Na2S2O3 (0.025)
8: molecular weight of oxygen
14. Half an hour later, a second water sample was taken in the same way as the
first water sample and titrated using the Winkler method. Then the final
dissolved oxygen level (Otak / cO2f) was determined with the same formula.
15. Fish are weighed to find out the volume of the tube after being reduced by the
volume of fish. The tube volume can be measured by the formula:
V = large respirometer - volume of fish
16. Fish oxygen consumption is measured by the following formula:
VO2 = (cO2i - cO2f) x V x H-1 x W-1
Description:
VO2: Oxygen consumption (mg/L/ hour)
cO2i: Initial dissolved oxygen (mg/L)
cO2f: Final dissolved oxygen (mg/L)
V: Volume of tube after reduced volume of nilem (V)
H: The time interval for the initial and final oxygen measurements (L)
W: Fish weight (g)
17. After finishing the observation, the respirometer is reopened, the circulation
pump and aerator are turned on.
 III. RESULT AND DISCUSSION

A. Result

Table 3.1. The Result of Observation of Aquatic Animal Respiration

Entourage VI

Hou Volume Weigh CO2i CO2f VO2

Group Fish
r (H) (ml) t (gr) (mg/L) (mg/L) (mg/L/H)
1 Big nile 0,5 130 124 5,6 5,6 0
tilapia
2 Big nile 0,5
tilapia
3 Big nile 0,5 9005 131 7,4 7 0,110
tilapia
4 Small nile 0,5 5435 26 6,4 4,8 1,337
tilapia

Calculation of Group 3:

1000
Ota (CO2i) = xpxqx8
100
1000
= x 3,7 x 0,025 x 8
100

= 7,4 mg/L
1000
Otak (CO2f) = xpxqx8
100
1000
= x 3,5 x 0,025 x 8
100

= 7 mg/L
VO2 = (CO2i - CO2f) x V x H-1 x W-1
(7,4 𝑥 7) 𝑥 9055
= 𝐻𝑥𝑊
(7,4 𝑥 7) 𝑥 9055
= 0,25 𝑥 131
0,4 𝑥 9055
= 0,25 𝑥 131

= 110,59 ml
= 0,110 mg/L/Hour
B. Discussion

Based on the lab activity, it was found that the weight of each fish are
different and also resulting different VO2 value. Big nile tilapia have weighs as
much as 124 gram that have VO2 value amount 0 mg/L/hour, 20 gram that have
VO2 value amount -0.694 mg/L/hour, and 131 gram that have VO2 value amount
0.110 mg/L/hour. For small nile tilapia fish have weigh as much as 26 gram that
have VO2 value amount 1.337 mg/L/hour. Based on data, big nile tilapia fish have
VO2 value smaller than small nile tilapia fish VO2 value. This result is accordance
with reference that lower-weight aquatic organisms (fish) will need more and use
oxygen in their lives than large fish. This is because small fish are more active to
move and are also useful for smooth metabolism (Fathuddin & Liestati, 2002).
Metabolism is a change or all chemical and energy transformations that
occur in the body. Metabolism is a highly coordinated cell activity, has a purpose
and includes various collaborations of many multi-enzyme systems. Metabolism
has four specific functions, among others, to obtain chemical energy from the
degradation of energy-rich food extracts or from solar energy, to combine these
building units into proteins, nucleic acids, lipids, polysaccharides and other cell
components, and to form and degrade biomolecules needed in special cell
functions (Putra, 2015). Metabolic rate is usually estimated by measuring the
amount of oxygen consumed by living things per unit time. This is possible
because oxidation of foodstuffs requires oxygen (in amounts known) to produce
energy that can also be known in number. However, the metabolic rate is usually
sufficiently expressed in the form of oxygen consumption (Suharsono et al., 2018).
The rate of metabolism is closely related to respiration because respiration is the
process of extracting energy from food molecules that depend on the presence of
oxygen. The metabolic rate is usually sufficiently expressed in terms of the rate of
oxygen consumption (Putra, 2015). The increase in temperature is related to the
metabolic system, rising temperatures will increase the metabolic rate. At the time
of the metabolic rate increased, oxygen demand and carbon exchange increased. If
at that time the blood does not contain enough oxygen to meet its needs, the animal
will experience hypoxic conditions or even asphyxia (there is no oxygen in the
body tissues) (Malini & Muliani, 2016).
 Winkler method is a way to determine the amount of oxygen dissolved in
water. In this method, the level of oxygen in water is determined by titration.
Titration is the addition of a known solution concentration (standard solution) into
another solution which is not known to be concentrated gradually until equilibrium
occurs (Chang, 1996). The Winkler method converts manganese(II) hydroxide
into manganese(III) hydroxide by quantitatively consuming dissolved oxygen. The
manganese(III) hydroxide is then used to oxidize iodide ion, which can then be
accurately measured by titration with thiosulphate. The simplicity and accuracy of
this titration allowed indirect calorimetry to be performed without sophisticated
gas analysis equipment. Thus, indirect calorimetry using Winkler titrations began
to supplant manometric techniques and direct calorimetry when investigators
measured aerobic metabolism of intact, aquatic animals. Winkler method then used
to assessing aerobic metabolic activity of aquatic ectotherms (Nelson, 2016).
In our lab activity we used nile tilapia fish as materials because
the salt tolerance / sanitaty is very high. Nile tilapia fish are not only freshwater
fish, these fish are also often found alive and rapidly developing in brackish waters,
such as brackish. Nilem fish can live in deep and wide waters and in narrow and
shallow ponds. Tilapia can also live in rivers that are not too heavy, in reservoirs,
swamps, rice fields, brackish water ponds, or in floating nets in the sea. Also nile
tilapia fish is cheap and easy to be cultivated (Schmidt, 1990).
According to Alaerts & Santika (1987), the steps of the titration method by
means of Winkler are first, the water sample was put into a 125 mL Winkler bottle
with the condition there was no air entering the sample. MnSO4 and KOH-KI
solution are added as much as 0.5 mL into Winkler bottle that has been contained
with water. 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). The water in the
Winkler bottle is reacted with H2SO4 as much as 0.5 mL, then shaken so that the
sediment becomes dissolved and formed yellowish liquid and left for 10 minutes.
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. The Winkler method
is done twice to get the average value. Soluble O2 formula: Ota = 1000/100 × p ×
q × 8.
 Oxygen consumption can be influenced by several factors, namely the intensity
of oxidative metabolism in cells, the rate of exchange that controls the movement
of water around the diffuse gills through it. Internal factors, namely the speed of
blood circulation and the volume of blood carried into the gills and the oxygen
affinity of hemoglobin, nutrition, disease, reproductive status and stress and
hormonal influences from these animals also influence oxygen consumption
(Lagler, 1977). 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 (Zonneveld et al., 1991).
Other factors that affect oxygen consumption in fish include: activities that
are fish with high activity, such as fish that are actively swimming will consume
more oxygen than fish that are not active. Age, which young fish will consume
more oxygen than older fish. Temperature, which fish at high temperatures is also
have high metabolic rate so that more oxygen consumption. Gender, where females
do more respiration because females have a more complex hormonal system than
males. Stress, which is 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). Oxygen consumption of fish also depends on oxygen solubility in
water, while the solubility of oxygen is influenced by salinity and temperature.
Oxygen solubility will decrease with increasing temperature and salinity, and the
amount of oxygen will vary according to variations in these parameters. During
the summer in the brackish area vertical vertical salinity stratification can occur
and a thermocline is formed (Malini & Muliani, 2016).
 IV. CONCLUSION

Based on the result can be concluded that are :

1. Oxygen consumption in big nile tilapia fish are 0.110 mg / L / hour, 0 mg / L /

hour, and -0.694 mg / L / hour, whereas in large tilapia fish it is 1.337 mg / L /
hour.
2. The initial dissolved oxygen value is 7.4 mg / L when the aerator is still on,
but when the aerator is turned off the final dissolved oxygen value drops to 7
mg / L. This shows a decrease in dissolved oxygen because fish with low
activity will consume less oxygen which is a response to the environment and
fish is get stresses.
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