OSMOREGULATION

By:
Name
NIM
Group
Cluster
Assistance

: Gibran Muhammad Tri R

: B1K014025
:3
:7
: Iis Islamiyah

PRACTICUM REPORT OF ANIMAL PHYSIOLOGY II

MINISTRY OF RESEARCH TECHNOLOGY AND HIGHER

EDUCATION
JENDERAL SOEDIRMAN UNIVERSITY
BIOLOGI FACULTY
PURWOKERTO
2015

I. INTRODUCTION

1.1

Background
Osmoregulation is a process to maintain the balance between amount of

water and diluted substance which contain in the body. Animals do the
osmoregulation process to balance their body concentration and their
environment. If a cell in animal body receive high level of water then the cell will
lysis, while if the cell receive level of water it will shriveled and die. The main
process of osmoregulation is called osmosis or water movement from high
concentration to low concentration. If value of osmoregulation capacity closing
two, so the fish will be grouped into hiperosmotic, while value of osmoregulatioc
capacity is about one, so the fish will be grouped into isoosmotic and if value of
osmoregulation capacity is bellow one, so the fish will be grouped in hipoosmotic
(Soegiri, 1988).
Osmotic pressure in body fluid is controlled in constant status so
physiological process inside the body can normally run that will be execute by
osmoregulation. Osmoregulation is a physiological function which need energy by
absorbing ion that goes through gill in fish and continue to excess the salt ion so
the fluid osmotic pressure remain constant. Osmoregulation process varie based
on temperature, season, age, physiological condition, gender and genotype
variation. The organ which have role in osmoregulation procress are kidney, gill,
skin, mouth membrane, and other specific organ which used by different type of
animals (Kurniawan, 2015).
Animals with limited tolerance against the variety of environment is called
stenohalin. While the animals with great tolerance against the variety of
environment is called eurihalin. Beside stenohalin and eurihalin, animals also can
be grouped based on pattern changes in their internal body against osmotic fluid
concentration as response to external variety (Sambasivia, 1978).

1.2

Objective
This experiment aim to study osmoregulation in euryhaline animals

(animal that can live in extensive water salinity), tilapia (Oreochromis niloticus)
and stenohaline animals, nilem fish (Osteochilus hasselti), and crab.

II. MATERIALS AND METHODS

2.1

Materials
The materials used in this lab are nilem fish seed (Osteochilus hasselti),

tilapia fish seed (Oreochromis niloticus), crab, water with (0 ppt, 5 ppt, 10 ppt, 15
ppt, 20 ppt, and 25ppt) salinity and EDTA.
The tools used in this lab are 1cc syringes injection, micropipette tip, label,
pipette, capillary hematocrit or eppendorf tube and paper disc.
2.2

Methods

2.2.1. Salinity tolerance observation

1.

Make medium water with 0 ppt, 10 ppt, 20 ppt and 30 ppt salinity each
for 4 liters.

2.

Medium is divided into 16 containers and each experiment consisted of

four experiment containers. Each container is labeled in based on it
salinity.

3.

Take into all experiment containers each 10 tilapia seed with direct
transfer method.

4.

Take 10 tilapia seed into experiment containers with lowest salinity.

5.

Take into all experiment containers each 10 Nilem fish seeds with direct
transfer method.

6.

Take 10 Nilem fish seeds into experiment containers with lowest salinity.

7.

Observe and record each death fish after 10, 20, 30 and 40 minutes for
direct transfer method.

8.

Observe and record each survived fish whenever the fish is moved from
0, 10, 20, and 30ppt after 24, 48, 72, and 96 hours.

9.

Survived fish data collected and then count it with formula

Where :

?? =

??
? 100%
?0

SR is degree of survival fish

Nt is number of survive fish in the end of observation

N0 is number of fish when the observation started.

2.2.2. haemolymph osmolality measurement in crab

1.

Take blood samples of Tilapia (with capillary hematocrit) that already

acclimated in medium salinity for 24 hours.

2.

Centrifuge the blood to obtain blood plasm.

3.

Measure plasma osmolality and medium osmolaltity with vapor pressure

osmometer.

4.

Calculate the ration between medium osmolality and plasma osmolality.

5.

Note all the data collected

2.2.3. Haemolymph osmolality measurement in crab

1. Coat the tip of syringe injection with EDTA so the haemolymph not
coagulated
2. Take samples haemolymph from crab leg section closed to crab body
using 1 ml syringe injection.
3. Measure haempolymph osmolality using vapor pressure osmometer.
4. Calculate the ration between plasma osmolality and medium osmolality
5. Note the data collected.

3.2

Discussion
Osmoregulation is the ratio between the value osmolarity plasma and

osmolarity medium. The osmolarity medium is a key determinant of osmotic work

rate which experienced by euryhalin animals. The greater the osmolarity of
medium is mark of high salinity. This caused by the increasing of dilluted ion
concentration. The higher the number of ions dissolved in the medium the
osmolarity fluid also is also higher. Salinity is one of many abiotic factor which
control aquatic organism survival rate (Wang, 2008).
Osmoregulation is a fundamental prerequisite for life in complex organisms.
Teleost fishes inhabiting a freshwater environment are at constant risk of
excessive hydration and electrolyte loss across body surfaces. To counteract this
tendency, FW-acclimated fish actively uptake ions across the gill and gut and
eliminate excess water by producing dilute urine. By contrast, osmoregulation for
teleosts in seawater is facilitated through the active extrusion of monovalent ions
by the gill and the conservation of water through re-absorption by the
gastrointestinal tract (Moorman, 2014).
Osmoregulation

divided

into

two,

there

are

osmoregulator

and

osmoconfomer. Osmoregulator is an animal which maintain it osmolality without

depend on the environment. The ability to regulate makes osmoregulator animals
can live in low osmolarity level environment such as freshwater, for example
fresh water shrimp. An osmoregulator animal if in hipoosmotic environment need
to dispose excess water, while if in hiperosmotic environment will constantly pick
up water to solve osmotic lose. Osmoconformer is an animal which have internal
osmolarity same with it environment so there will no tendendention to pick or lose
water. Most of osmoconformer animals live in environment with stable chemical
composition such as salt water so it have constant osmolarity. Most
osmoconformer is sea invertebrate like jellyfish, seashell, and crabs (Isnaeni,
2006).
The term stenohaline is used for animals which can live only within a
limited range of outside concentrations. Stenohaline animals do not have the
mechanisms to maintain a gradient between the extracellular fluid and the media.
Most estuarine crustaceans are euryhaline and exhibit a hyper-isosmotic pattern of

regulation; maintaining haemolymph hyperosmotic at low salinities by active

uptake of ions but conforming at higher salinities, For example gold fish
(Carassius auratus auratus) (Campbell, 2011). Beside stenohaline, there also
euryhaline animals. euryhaline animals can live over a wide range either by
toleration of environmental salinities. How the microbiota in these two host
species respond to environmental changes is an important question not only to
researchers in aquaculture physiology but also to the fish industry, for example
Tilapia fish (Oreochromis niloticus) that can undergo osmoregulation process in
0-35% salinity (Kurniawan, 2015).
Based on osmotic concentration, a fluid can be divided into hypoosmotic,
isosmotic and hiper osmotic. Hypoosmotic is a fluid which the osmotic
concentration is lower then the environment concentration. Isoosmotic is fluid
which the osmotic concentration same as the environment concentration.
Hierosmotic is fluid which the osmotic concentration is higher then the
environment (Karim, 2007).
Osmoregulation in every type of fish is different based on their
environment where the fish live. For fresh water fish because they live in low
salinity water, they will excess the high concentrated urine which basically
contain high level sald that makes the urine concentrated. to balance osmorality
state between their body and their environment. While for saltwater fish they will
excess the low concentrated urine which basically contain less salt, so they can
balance their body and environment osmorality state (Gordon, 1982).
Based on Karim (2007) when the fish is exposed in danger or not in their
natural environment before taking it blood sample, their heart rate will change and
will causing lysis after the blood has been taken. The circumspection while taking
blood also decreasing the possibility lysis in blood sample.
Based on observation group 1 using Tilapia fish for their survival rate by
Direct Transfer method in salinity 0 ppt which observed for 40 minute the
survival rate of tilapia fish is 100%, in salinity 10 ppt which observed for 40
minute the survival rate of tilapia fish is also 100%, in salinity 20 ppt after 30
minute observation the survivial fish rate is 80% which remain until 40 minute, in
salinity 30 ppt after 30 minute observation the survival fish rate is 80% and in 40

minte the survival fish rate decrease again until 70%. While in group 2
observation which using Nilem fish, their survival rate is more lower than Tilapia
fish where in salinity 0 ppt after 40 minute the survive rate remain 100%, in
salinity 10 ppt after 40 minute observation the survival rate also 100%, while in
salinity 20 ppt after 10 minute observation the survival rate is 90% and decreasing
in 20 minute to 60%, in 30 minute 10% and 40 minute 0%, and finally in salinity
30 ppt the first 10 minute the survival rate already down to 50%, in 20 minute
became 40%, in 30 minute decreasing again into 30% until 40 minute observation.
Based on the observation above we can conclude that Tilapia fish is euryhaline
animal which can live in wide range toleration salinity, while Nilem Fish is
stenohalyne animal which can only live in limited range toleration salinity. For
gradual transfer group 1 observation in Tilapia fish in 24 hours in 0 ppt salinity
fish survival rate is 100% also in 10 ppt salinity to next 48 hours the survival rate
remain the same, after moved into 20 ppt salinity for another 24 hours the fish
remain 40% and finally in last 24 hours the fish remain 10%. While for group 2
observation using Nilem fish by gradual transfer method for first 24 hours in 0 ppt
salinity the fish still in 100% survival rate, when it moved to 10 ppt salinity for
another 24 hours it survival rate decrease to 90%, in 24 hours in 20 ppt salinity the
fish drop drastically into 30% and in last 24 hours in 30 ppt salinity all fish is dead
which makes it survival rate become 0%. this also related with Cambell (2011)
statement which said that Tilapia is saltwater fish who can live in wide tolration
salinity, while Nilem is freshwater fish who can only live in specific toleration
salinity.
The results for measurement of plasma osmolality and medium salinity 0
ppt, 5 ppt, 10 ppt, 15 ppt, 20 ppt, 25 ppt, and 30 ppt, its osmotic media
concentrations consecutively are 247 mmol/kg, 490 mmol/kg, 529 mmol/kg, 481
mmol/kg, 582 mmol/kg, 576 mmol/kg, and 806 mmol/kg, the osmotic blood
plasma concentrations of Tilapia fish consecutively are 545 mmol/kg, 461
mmol/kg, 498 mmol/kg, 832mmol/kg, 556 mmol/kg, 336 mmol/kg, and 560
mmol/kg. while the osmotic blood plasma concentrations of Nilem fish in a row
are 545 mmol/kg, lysis, lysis, lysis, 463 mmol/kg, lysis, and 560 mmol/kg. The
result of hemolymph osmotic concentration of crab consecutively are 545

mmol/kg, 839 mmol/kg, 576 mmol/kg, 518 mmol/kg and 474 mmol/kg, 383
mmol/kg, and 560 mmol/kg. Hickman (1972) adds that tilapia is a freshwater fish
that can tolerate salinity changes in the medium. When the fish are in the medium
which are not freshwater, then there is a changes in osmoregulation where the
water in the medium will enters the body at freshwater medium and body fluids
come out in a liquid medium that arent freshwater, then at high salinity medium
fish should be able to keep fluids in its body and pulled out ions that enters the
body when the fish drink water from the media

IV.

CONCLUSSION

Based on the result can be concluded that :

1. Osmoregulation is the ability organism ability to maintain the equilibrium
level in body
2. Tilapia is included in euryhaline animals, animals which can live in waters
with wide salinity tolerance range, while Nilem fish is included in
stenohalin animals, animals wich can live in waters with narrow/specific
salinity range.
3. The higher the concentration of the media, the higher the concentration of
the blood plasma..

REFERENCES.
Campbell, N. A., Reece, J. B., Mitchell, L. G. 2011. Biologi 9th edition. Erlangga,
Jakarta.
Gordon, M. S. 1982. Animal Physiology Principles. MacMillan Pub. Co., New
York.
Hickman, C. F. 1972. Biology of Animals. The C. V. Mosby Company, Saint
Louis.
Isnaeni, Wiwi. 2006. Fisiologi Hewan. Kanisius, Yogyakarta.
Karim, Y. M. 2007. The Effect of Osmotic at Various Medium Salinity on
Vitality of Female Mud Crab (Scylla olivacea). Universitas Hasanudin,
Makassar. Vol.14. No.1.
Kurniawan, D.Dwi, S. 2015. PENGENDALIAN Saprolegnia sp. PADA IKAN
NILA (Oreochromis niloticus) DENGAN SALINITAS AIR YANG
BERBEDA. Program Studi Manajemen Sumberdaya Perairan, Fakultas
Pertanian Universitas Sumatera Utara, Part B (2015) 1-9.
Moorman, B. P. 2014 The osmoregulatory effects of rearing Mozambique tilapia
in a tidally changing salinity. Hawaiian Institute of Marine Biology,
General and Comparative Endocrinology 207 (2014) 94102
Sambasivia. 1987. Ictyology. John Wiley and Sons Inc, New York.
Soegiri. 1988. Zoologi Umum. Erlangga, Jakarta.
Wang, Pei-jen et al,. 2008. Differential Responses in Gills of Euryhaline Tilapia,
Oreochromis mossambicus, to Various Hyperosmotic Shocks.
Department of Life Sciences, National Chung-Hsing University, Taichung,
402, Taiwan. Comparative Biochemistry and Physiology, Part A 152
(2009) 544551.