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AQUATIC ANIMAL RESPIRATION

Name : Nahdlini Salma Sabila


Student ID : B1B017002
Entourage : VI
Subgroup :1
Assistant : Nisa Baiti

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

Fish are aquatic animals that consume oxygen dissolved in water. oxygen
uptake in fish is done by the main respiratory organs ie gills. The amount of
oxygen uptake through gills can be measured by static water or running water
methods. The oxygen consumption parameter is used to assess the fish
metabolism rate because most of the fish source energy comes from aerobic
metabolism. To perform aerobic metabolism, fish need to consume oxygen.
Therefore, changes in fish oxygen consumption can be used to assess changes
in metabolic rate. The metabolism of the poikioterm animals is influenced by
changes in ambient temperature. When temperatures are low, metabolism falls
and metabolism increases at elevated ambient temperatures (Sudibyo, 1999).
Respiration is the process of taking O2 from the environment into the
animal body and removing CO2 from the body to the environment. Respiration
in aquatic animals, examples of fish include extraction or taking O2 from the
waters (Yuwono, 2001). Dissolved oxygen is required by all living bodies for
respiration, metabolism or exchange of substances which then generate energy
for growth and culture. In addition, oxygen is also required for the oxidation of
organic and non-organic materials in aerobic processes. The main source of
oxygen in a waters comes from a diffusion process of free air and
photosynthetic processes of organisms living in these waters (Asmawi, 1983).
Poikilotherm animal's metabolism is affected by changes in environment
temperature. At low temperatures the metabolism decrease and will increase
when the environmental temperature increases. The metabolic rate is inversely
proportional to the concentration of dissolved oxygen and correlated with
oxygen consumption and blood hemoglobin synthesis. When the oxygen
concentration is low and the temperature increases, the rate of metabolism
increases while when the oxygen concentration high and low temperatures, the
low metabolic rate (Sabandiah, 1998).
B. Purpose

The purpose of this experiment are:


1. To calculate the rate of oxygen consumption in aquatic animal.
2. Students can measure the oxygen consumption of aquatic animal buy
using titration (Winkler method)
3. Students can measure the metabolic response of aquatic animal related
with the body weight as well as environmental changes or stress.
II. MATERIAL AND METHOD

A. Material

The tools used in this laboratory activity are aerator, technical scales,
large measuring cup, respirometer, Winkler bottles, erlenmeyer, burette, and
statif.
The materials used in this laboratory activity are nilem fish
(Osteochillus vittatus), nila fish (Oreochromis niloticus), KOH-KI solution,
H2SO4 solution, Na2S2O3 solution, MnSO4 solution, and amylum reagent.

B. Method

The method used in this practical are :


1. Respirometer was run, the water pump is turned on for 15 minutes.
2. The weight and the volume of the fish was measured (Vfish = V total –
V1).
3. The fish was put into respirometer, make sure there is no bubble inside.
4. The water sample was taken to Winkler bottle 250mL.
5. The solution of MnSO4, KOH-KI, and H2SO4 was added by 1 mL each
solution.
6. The solution was taken to Erlenmeyer as much as 100mL and the amylum
was added for 2-3 drops.
7. The early titration was started by Na2S2O3.
8. The water sample was taken to Winkler bottle as much as 250mL after 15
minutes.
9. The final titration was done.
10. The consumption of O2 was calculated by
1000
𝑂𝑡𝑎 = ×𝑝×𝑞×8
100
Information:
Ota: initial dissolved oxygen (mg / L)
p: used Na2S2O3 solution
q: normality of Na2S2O3 (0.025)
8: molecular weight of oxygen
11. Half an hour later the second water sample is taken in the same way as the
first sample and titrated with the Winkler method to determine the final
dissolved oxygen (Brain) level measured as the Ota formula.
12. The oxygen concentration (mg / g / hour) of fish is measured using the
Fidhiany method with the following formula:
𝑉𝑂2 = (𝑐𝑂2𝑖 − 𝑐𝑂2𝑓 ) × 𝑉 × 𝐻 −1 × 𝑊 −1
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)
13. After completing the observation, the respirometer is reopened, the
circulator and aerator are turned on.
III. RESULT AND DISCUSSION

A. Result

Table 3.1. The Result of Observation of Aquatic Animal Respiration


Entourage VI
H Volume Weight CO2i CO2f VO2
No. Fish (mg/L/hour)
(hour) (ml) (g) (mg/L) (mg/L)
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 5465 269 6,4 4,8 1,337

Calculation of Group 1:

1000 1000
𝑂𝑡𝑎 = ×𝑝×𝑞×8 = × 2,8 × 0.025 × 8 = 5.6
100 100

1000 1000
𝑂𝑡𝑎𝑘 = ×𝑝×𝑞×8 = × 2,8 × 0.025 × 8 = 5.6
100 100

𝑉𝑂2 = (𝑐𝑂2𝑖 − 𝑐𝑂2𝑓 ) × 𝑉 × 𝐻 −1 × 𝑊 −1


(5.6 − 5.6) x 9045 𝑚𝑔
= =0 ⁄𝑔/ℎ
0.25 𝑥 124
B. Discussion

Based on the results of experiments that have to be got results obtained after
the experimental measurement of oxygen consumption in large nilem fish, in the
first calculation obtained initial amount of dissolved oxygen (cO2i) of 5,6 mg/L
and the second amount of dissolved oxygen end (CO2f) of 5,6 mg/L. After
calculating of average obtained results of the dissolved oxygen water of large
nilem fish is 0 Mg/L. Oxygen consumption obtained from the calculation results
show that the dissolved oxygen is the same than the beginning of the end of
dissolved oxygen. This is in inconsistent with the statement Barnes (1965), which
states that the fish get oxygen from the oxygen dissolved in the water. This might
happened due to to human error while do the titration, and th not significant time
between the beginning and the end calculation. The fish that is used are nilem fish
(Osteochillus vittatus) and nile tilapia fish (Oreochromis niloticus). The use of
those fish in this laboratory activity is because the fish are easily obtained, and
live in freshwater environments with a sufficient range of dissolved oxygen
content so that the respiratory system is more effective (Cholik et al., 2005).
Oxygen consumption is one of the physiological parameter that can be used
to estimate the rate of metabolism indirectly, namely by measuring the rate of
oxygen used in the oxidation process. The consumption of oxygen can be used as
an approach for estimating metabolic practical because of the amount of heat that
is generated for each liter of oxygen used in the metabolism of almost constant
regardless of what is in the oxidation of whether fats, proteins and carbohydrates.
Such a process would generate a further energy will be used to sustain life,
including environmental adaptation (osmoregulation) (Affandi & Tang, 2002).
Metabolism constitutes a fundamental property of all organisms. Metabolic rate is
commonly described to scale as a power function of body size and exponentially
with temperature, thereby treating the effects of body size and temperature
independently. Mounting evidence shows that the scaling of metabolic rate with
body mass itself depends on temperature (Ohlberger et al., 2012).
Oxygen is very influential in the body's metabolic processes. This
oxygen demand is directly proportional to the metabolic rate. Metabolic rate
can be calculated by measuring the amount of oxygen consumed by an
organism per unit time. This is possible because the oxidation of food
ingredients requires oxygen to produce known amounts of energy as well.
However, the metabolic rate is usually sufficiently expressed in terms of the
rate of oxygen consumption (Suharsono et al., 2018). Measurements of oxygen
consumption of fishes and other aquatic organisms have been made over time
using a variety of respirometry techniques such as closed respirometry, flow-
through (or open) respirometry, and intermittent-flow (or stop-flow)
respirometry. In this practical class, The experiments were carried out using
closed respirometry. Closed respirometry-system involves placing an animal in
a closed (sealed) metabolic chamber and measuring the decline in oxygen
concentration in the chamber over time (Svendsen et al., 2016).
Oxygen consumption rate can be determined in various ways, such as by
using microrespirometer, Winkler methods, as well as respirometer Scholander.
The use of each way based on the type of animals to be measured rate of
oxygen consumption. Mikrorespirometer used to measure oxygen consumption
of small animals such as insects or spiders. Scholander respirometer used to
measure the rate of oxygen consumption of animals such as frogs or mice.
Winkler method is a way to determine the amount of oxygen dissolved in the
water. In this method, oxygen levels in the water were determined by titration.
Titration is the addition of a solution of known concentration (standard
solution) to a solution of another unknown concentration gradually until there
is equilibrium. With Winkler method, we can determine how much oxygen is
consumed by aquatic animals such as fish (Chang, 1996).
Measurement of oxygen consumption rate by the method of Winkler,
Winkler using 250 ml bottle. Water aeration results that have been filled with
tilapia accommodated in the bottle Winkler then added with a solution of KOH-
KI, MnSO4 and H2SO4. The function of the aqueous KOH-KI is to bind oxygen,
the function of MnSO4 is to bind oxygen, and the function of H2SO4 solution is
to neutralize the KOH-KI and MnSO4. The water inside the bottle Winkler then
put into the Erlenmeyer flask, then drip with starch and titrated by using a
solution of Na2S2O3. After a few drops of starch solution will change color to
dark blue, then titrated with a solution of Na2S2O3 until clear. The function of
the starch is as an indicator of O2 while the functions of Na2S2O3 solution is as
titration (Chang, 1996).
According to Wetzel & Linkens (2000), the function of the solution
used in the titration process including:
1. MnSO4 solution and KOH-KI: to form a precipitate brown turbid, indicating
that there are still O2 in the sample. If the resulting white precipitate then no
O2 dissolved in the sample.
2. H2SO4: to change the solution initially murky brown to brown translucent
(yellowish). This solution is not formed from the reaction between sulfuric
acid and manganese oxide form of manganese sulphate.
3. Starch: as an indicator of the change clear solution is brown to purple
(violet).
4. Na2S2O3: titration as p values to look for levels of dissolved O2.
Temperatures were included in the most influential environmental factors
are the metabolic rate, otherwise the concentration of dissolved oxygen is a
factor lower than the rate of metabolism.
The amount of oxygen consumption is influenced by several factors,
including body weight or size, volume, age, activity of the fish, and the
environmental temperature. This is according to a statement Gordon (1972), that
the size or weight of the fish affects the consumption of oxygen for metabolism.
The smaller the weight of the fish, the greater the rate of oxygen consumption.
The rate of metabolism in fish affected by the increasing body weight, the
volume of fish, the fish activity, ambient temperature, and salinity so that the
greater oxygen consumption (Ville et al., 1988).
The level of oxygen consumption is influenced by two factors, they
are external and internal factors. External factors affecting the concentration of
dissolved oxygen, temperature, salinity, light, status of food and carbon dioxide,
while the internal factor is the species, size (stadia), activity, sex, reproduction
and molting (Karim & Syahrul, 2006). Weight and volume of the body's oxygen
consumption is influenced by the activity of the fish itself. Fish with a small
volume and weight of the body has a great activity when compared to the
volume of fish and a large body weight, so the volume and weight of the fish
with a small body to consume more oxygen (Yuwono, 2001).

According to Lagler (1977), O2 consumption can be influenced by


several factors, they are:
1. The intensity of oxidative metabolism in cells.
2. Free exchange which controls the movement of water around the gills that
diffuses through it.
3. Internal factors are the speed of blood circulation and blood volume that is
brought to the gills.
4. Oxygen affinity of hemoglobin.
IV.CONCLUSION

Based on the result and discussion, can be concluded that the oxygen
consumption rate of fish group 1 entourage VI is 0 mg / g / hour, due to the same
result in the earlier calculation and final calculation.
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