Bicol University

College of Science
Department of Biology

LABORATORY ACTIVITY 2
OSMOREGULATION

Date performed:

January 18, 2011

Performed by:

Zarieh Dawn L. Novela

BS Biology 3A
INTRODUCTION
The earthworm belongs to the phylum Annelida. This group of higher invertebrates
possesses a number of highly developed organs and organ-systems for performing various vital
functions. One of the most important is nephridia where in one of its function is the regulation of
osmotic pressure in the body in order to maintain constant water.

OBJECTIVES
• Observe the effect of salinity on changes in the volume and weight of earthworms
• Use the earthworms as model for explaining basic osmoregulatory mechanisms

MATERIALS AND METHOD

I. Preparation
A. Earthworm
1. Collect at least 30 earthworms (of more or less the same size) for this experiment. The
best time to hunt for worms is on a damp night when it is not too cold. Use a flashlight to help
locate them in patches of grass, in parks or fields, or in lawns. You may have plenty in your
garden. Usually, you will find them stretched out in the surface of the ground with their posterior
ends inside their burrows. They may pop back into the ground when you appear.
2. If you collect during the day, you may dig for them preferably at places where the soil
is loose. Digging for worms, however, is not equally successful in the different parts of the
garden or field. There are some areas where one will find many worms and others where worms
will be quite scarce. It is preferable to dig for worms after heavy rain. Heavy rain drives the
worms to the surface where they may be seen crawling on the ground.
Actually, worms come up from their flooded burrows below the ground. It is also suggested that
you record your field observations of the worms’ behavior.
3. Bring the worms to the laboratory in an uncovered can or glass jar, or a box containing
some damp soil and some moist leaves or grass.
4. When you reach home, tumble them on paper towels to remove excess soil.
5. In groups of five, place them in separate containers with rain or aged tap water (with
the heavy chlorine removed). Time your preparation so that after the sixth hour, you are to start
with the experiment in the laboratory.

B. Reagents
• Six salt solutions of the following concentrations: 0.03 M, 006 M, 0.09 M, 0.12 M, 0.14
M and 0.15 M.
Note: you will be immersing the worms in these different media and observing for their
osmoregulatory responses. You will use five containers with five worms each. The worms in the
sixth container are extras for possible replacements if death takes place during the six hours of
immersion in tap water.

II. Experiment Proper

1. Weigh the worms in groups of five and place them in separate containers with aged tap
water.
2. Weigh at the end of 15 and 30 minutes. If the weights are fairly constant already,
determine the volume of each group by volume displacement in a graduated cylinder containing
water.
3. After you have determined the weight and volume of each group, transfer each group
to a different dish containing one of the salt solutions.
4. Weigh the worms at 20-minute intervals within the 100 minutes of immersion. At the
end of the experiment, determine their volume.
5. Record your results in the table below.
6. Using graphs show the relationship between osmotic concentration of the medium and
the weight and volume changes exhibited by the animals throughout the duration of the
experiment. For this, plot weight or volume change against time with a color or symbol legend
for the different experimental groups.
7. Similarly, make graphs to depict the relationship between percentage weight change or
percentage volume change through time and the osmotic concentration of the medium.

RESULTS AND DISCUSSION

We have collected earthworms and grouped it into 5 and placed it in different containers
with different salt concentration. This was to observe the osmoregulatory response of
earthworms.
0.09 M NaCl
Moles of solute = (molarity)(liter of solution)
= (0.09 M) (0.05 L)
` = 0.0045 moles of NaCl

Mass of NaCl = (0.0045 moles NaCl) (1 mole NaCl / 58.44 g NaCl)

= 0.00007.7 g of NaCl

*0.06 M NaCl *0.03 M NaCl

M1V1 = M2V2 M1V1 = M2V2
V1 = M2V2 / M1 V1 = M2V2 / M1
= (0.06 M)(50 ml) / (0.09 M) = (0.03 M)(50 ml) / (0.09 M)
= 33.33 ml = 16.67 ml
*0.012 M NaCl *0.015 M NaCl
M1V1 = M2V2 M1V1 = M2V2
V1 = M2V2 / M1 V1 = M2V2 / M1
= (0.012 M)(50 ml) / (0.09 M) = (0.015 M)(50 ml) / (0.03 M)
= 6.67 ml = 8.33 ml

0.03 M 0.06 M

0.09 M 0.012 M

When the earthworms were placed in different media,

0.012 M
we observed that they furiously wriggle out of the Petri
dish as if they were being drowned.
Table 1. Weight of earthworms while immersed in media of varying salinity
Length of Weight of worms in media or varying concentration
exposure, min 0.03 M 0.06 M 0.09 M 0.12 M 0.15 M
0 6.12 g 6.12 g 6.12 g 6.12 g 6.12 g
20 6.53 g 6.97 g 5.75 g 6.65 g 6.12 g
40 7.21 g 6.77 g 5.73 g 6.35 g 5.97 g
60 7g 6.22 g 5.71 g 6.25 g 6g
80 7.6 g 6.18 g 5.65 g 6g 5.75 g
100 7.5 g 6.37 g 5.64 g 6g 5.75 g

Weight of earthworms while immersed in media of varying salinity

8

7.5
weight in grams

0.03 M
7
0.06 M

6.5 0.09 M
0.12 M
6
0.15 M
5.5
0 20 40 60 80 100

Table 2. Volume of earthworms in different media at start and end of experiment

Length of Volume of worms in media
exposure, min 0.03 M 0.06 M 0.09 M 0.12 M 0.15 M
0 3 ml 5 ml 5 ml 9 ml 6 ml
100 9 ml 3.5 ml 5 ml 4.6 ml 4.2 ml

Let us study the anatomy and morphology of an earthworm. Partitions divide their body
into a series of coelomic chambers, each with repeats of muscles, blood vessels, branching
nerves, and other organs. The gut extends through all chambers, from mouth to anus. As in all
annelids, an earthworm’s body is enclosed in a flexible cuticle of secreted material. The
permeable cuticle is good for gas exchange but not for conserving water. That is one reason
annelids cannot venture from aquatic habitats or moist habitats on land.
Annelids contain a system of nephridia, a series of units that control the volume and
composition of the fluids. Usually, each unit has a funnel that collects excess fluid from one
coelomic chamber and drains it into a small tube in the chamber behind it. The tubes lead to a
bladder, which delivers fluid to a pore at the surface in the body wall of the next coelomic
chamber.
 Looking at the graph above, it is observable that from the initial weight of 6.12 grams,
the weight of earthworms placed in 0.09 M, 0.012 M, and 0.015 M of salt solution decreases. In
contrast, those in 0.06 M and 0.03 M solution increases. 0.06 M and 0.03 M salt solution are
hypotonic solutions, that is, it has a lower concentration of solute than that of earthworms,
therefore, the water moves in the body of the organism resulting to the increase of its weight. In
the case of 0.09 M salt solution, there should neither decrease nor increase in the weight of the
earthworm since this is isotonic. On the other hand, 0.012 M and 0.015 M salt solution are
hypertonic solutions, that is, it has higher concentration of solute than that of the organism,
causing it to release water from its body. These are all for the balance of water content of the
organism.

Guide Questions:
1. What determines the salt concentration of the soil? What is the effect of rainfall on the
soil? On the behavior of earthworms? What mechanisms are of survival value to the
earthworms in variable environments?

2. What response in the weight and volume of the earthworms do the different salt solutions
evoke? Depending on the salinity of the media, the weight and volume of the earthworms
could increase, decrease or no change.

3. Compare the water exchanges in the worms exposed to varying saline media. What is the
regulatory role of these exchanges? The hypotonic solutions causes increase in weight
and volume in earthworm, while the hypertonic solutions causes decease in weight in
earthworm. There would be consistent weight and volume in earthworm in an isotonic
solution.

4. What are the osmoregulatory organs or structures of the earthworm? Nephridia

5. Why ionic regulation, volume regulation, and osmoregulation are inextricably associated
with each other? Those variables are directly proportional to each other. And also they
are regulated by the same organ of an organism, which is nephridium in earthworms.

6. Aside from osmoregulation, what is the other reason for the earthworm to crawl to the
surface at night? Do they have eyes?
They crawl to the surface at night taking advantage since the sun won’t dry them
therefore they will get enough oxygen. Earthworms don't have eyes, but they do have
light-sensitive cells scattered in their outer skin. These cells don't enable earthworms to
see images, or forms, but they do give their skin the capacity to detect light and changes
in light intensity.