BIOLOGY INVESTIGATORY

PROJECT

CRYPTOBIOTIC
DESICCATION
Submitted by-
Geetali
XII-A
R. no 7
 ||CERTIFICATE||

This is to certify that Geetali Ulhas Madkaiker, of

class XII-A has successfully completed her BIOLOGY
INVESTIGATORY PROJECT on the topic
“CRYPTOBIOSIS” as prescribed by, Mrs. Charanjit
Kaur, during the academic year 2020-2021 as per
the guidelines issued by Central Board of
Secondary Education- CBSE

Name of Examiner Name of Teacher

Signature of Examiner Signature of Teacher

PAGE 1
ACKNOWLEDMENTS:
I would like to express my special thanks and
gratitude to my Biology teacher Mrs. Charanjit
Kaur, as well as our principal Mrs. Jayshree
Venkatraman, who gave me the golden
opportunity to do this wonderful project on the
topic “CRYPTOBIOSIS”, which also helped me in
doing a lot of Research and I came to know
about so many new things I am really thankful
to them.
Secondly, I would also like to thank my parents
and friends who helped me a lot in finalizing
and compiling this project within the limited
time frame.

PAGE 2
CONTENTS:
1. AIM

2. INTRODUCTION

3. MATERIALS REQUIRED

4. PREPARATIONS

5. PROCEDURE

6. OBSERVATIONS

7. CONCLUSION AND ANALYSIS

8. PRECAUTIONS

9. BIBLIOGRAPHY
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 AIM:
To investigate the effect of various pH levels on development of organisms
showing Cryptobiotic Desiccation [Artemia].

 INTRODUCTION:
o What is Cryptobiosis:
 Cryptobiosis is a state of extreme inactivity in response to adverse
environmental conditions. The most common type of cryptobiosis is
Cryptobiotic Desiccation or drying out Examples of organisms with
cryptobiotic desiccation include nematodes, brine shrimp and a
majority of plant seeds.
o What are Brine Shrimps?
 Artemia or Brine shrimp are crustaceans that are classified in the
phylum Arthropoda. They live in saltwater lakes to avoid predators.
o Life Cycle of Artemia
 Female brine shrimps lay eggs, known as cysts. These cysts have the
ability to remain dormant till they have suitable hatching conditions.
These dormant cysts are biconcave and turn spherical when hydrated.
This ability of remaining in a state of extreme inactivity in response to
adverse environmental conditions is called Cryptobiosis
 The hatched shrimp larvae are called nauplii. These nauplii have a
different anatomical structure if
compared to an adult Artemia. The
nauplii shed their exoskeleton a few
hours after hatching and enter their
second larval stage. After a few more
sheds, they reach the mature or adult
stage. Hence it takes them about a week
or so from the time of hatching to reach
their mature state. Mature brine shrimp might grow to as much as half
an inch in length and live for up to three months.
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  These mature adult shrimp mate, and the female lays a batch of eggs
that will either hatch or undergo cryptobiosis based on the conditions
available.

o Hatching and Raising the shrimp:

 A simple funnel shaped apparatus can be used to hatch the eggs to
the first larval stage. To ensure maximum hatch rate aeration is very
important.

 An economical way to make this apparatus is to use an inverted

bottle with an air pump attached to it (the cap)

 Fill this apparatus with about one liter of water and add one to one
and a half teaspoons of non-iodized salt. After complete dissolution
of the salt add half a teaspoon of the eggs and place it in sunlight
with good aeration.

 After the eggs have hatched, separate then into different

observation chambers.

o Artemia and its relation to water pollution:

 Artemia or brine shrimp are found in saltwater bodies such as lakes.
Despite of their small size, these organisms play a very important
role in maintaining the ecosystem and food chain of saltwater lakes
and even the slightest change can lead to unfavorable conditions for
the other members of the food chain.

 These crustaceans thrive in optimal salt and pH levels in the water

and can be adversely affected by drastic changes in the same cause
by overheating due to global warming and water pollution due to
improper waste disposal.

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  The objective of this investigation is to find out the effects of these
changes on the development of these crustaceans.

 MATERIALS USED:
o Apparatus:
 A soda bottle.

 A tank (for observation of normal levels)

 Four clear containers

 An air pump

 A pipette

 A magnifying glass

 Dropper

o Chemicals and other Materials:

 5% Acetic acid

 Sodium bicarbonate (NaHCO3)

 Rock salt (or non-iodized salt)

 Activated Dry yeast

 Artemia cysts

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 PREPARATION:
o Artemia nauplii (freshly hatched):
 A simple funnel shaped apparatus can be used to hatch the eggs to
the first larval stage. To ensure maximum hatch rate aeration is
very important.

 An economical way to make this apparatus is to use an inverted

bottle with an air pump attached to it (the cap) or buy using a
clear container with an air pump.

 Fill this apparatus with about one liter of water and add one to one
and a half teaspoons of non-iodized salt. After complete
dissolution of the salt add half a teaspoon of the eggs and place it
in a spot with sunlight with good aeration.

 After the eggs have hatched into nauplii, separate then into
different observation chambers.

o Growing the nauplii in the observation tank:

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  Fill the observation tank with one liter if water and dissolve 1 ½
teaspoons of rock salt in it. Add a pipette full of freshly hatched
nauplii in this tank. Mark this as day one in the observation table.

 In order to nourish the nauplii, a simple mixture of yeast and water

is prepared by mixing a few balls of activated dry yeast in one
tablespoon of warm water. Mix thoroughly and let it cool to room
temperature. Now, using a dropper, add one to two drops
(depending on number of larvae) to the tanks every two to three
hours. (This mixture is to be prepared freshly only)

 PROCEDURE:

o Take the four clear containers (labeled A,B,C and D respectively) and fill
them with 800ml of water.

o Add ½ teaspoon of rock salt to container A. Stir this until completely

dissolved and add one pipette full of nauplii in it. Note this in the
observation table.

o Add 4 teaspoons of rock salt to container to container B. stir this until

completely dissolved. Note this in the observation table

o Add 2 teaspoons of 5% acetic acid to container C, and stir it in, and add
one pipette full of nauplii in it. Note this in the observation table

o Add 2 teaspoons of sodium bicarbonate in container D and stir it. Add

one pipette full of nauplii to this. Note this in the observation table
PAGE 8
 o Now, observe the size and number (if possible) of nauplii in each of these
containers and write it in your observation table.

o Continue observing the shrimps for the next two days, now add a pinch
of dry yeast to each one of the containers and the observation tank.

o Continue observing the growth and survival rate of the shrimps in each of
these samples and note this in the observation table.

 OBSERVATIONS:

Tank No. Change Salt level pH scale

made(impurity)
A Lower level of salt added Lower 8

B Higher level of salt added Higher 8

C 5% Acetic acid Normal 4

D Sodium Bicarbonate Normal 10

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o Day 1:
 All tanks have normal activity level

 All tanks were aerated for sufficient amount of time and ample
sunlight was provided.

 Growth is normal

o Day 2:
 C and D showed slightly less activity and a slightly lower number
of Artemia but growth level is normal

 A and B displayed less activity only and growth level was normal

 Shedding of exoskeleton noted in all tanks.

o Day 3:
 C and D’s activity further deteriorated, it is also noted that the
(now second naupliar stage) Artemia’s growth is slightly
retarded(in comparison with observation tank)

 A and B has reduced activity, growth level normal

o Day 4:
 Lower levels of activity noted in all tanks

 Only a handful of Artemia are seen in C and D, with an increasing

number of carcasses.

o Day 5:
PAGE 10
  All tanks have countable number of juveniles.

o Day 6:
 C and D are completely empty.
 A and B have eight and five left respectively.

o Day 7:
 All tanks except A are fully empty.

 PICTURES OF EXPERIMENT:
o Image of observation tank:

PAGE 11
Fig. 1: Observation tank and setup

o Image of different stages of Artemia:

Fig. 2.1: Different stages of Artemia

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Fig. 2.2: Artemia Juvenile

o Image of performed experiment:

Fig. 3.1: Labelling the tanks and adding the impurities

Fig. 3.2: Nauplii being attracted to light which aids in transfer

PAGE 13
 RESULT:
o Growth Rate
 The growth is optimal in case of normal salt level and a pH value of
8.
 The growth either completely retards or stops in case of
impurities.
o Life expectancy and survival rate:
 The life expectancy and survival is optimal in case of normal salt
level and a pH value of 8.
 Most Artemia cannot survive adverse pH values or salt levels.

 CONCLUSION AND ANALYSIS:

o Global warming:
 One of the major causative factors for changes in salt levels to take
place under normal circumstances is Global warming.
 Even changes if a few degrees can drastically change the salt levels
in water bodies; this change can disrupt the lives of Artemia, which
in turn can disrupt entire ecosystems, given that multiple aquatic
organisms and birds rely on them for food.
 One of the only ways this can be prevented is taking responsibility
as a society. Baby steps matter a lot, something as simple as
walking, cycling or carpooling to work/school, can make a huge
difference if done by a large number of people. Because at the end
of the day we are also just organisms on this planet.
o Pollution:
 One very concerning yet common reason for change in pH levels in
water bodies is pollution. Both air and water pollution can lead to
increased acidity or basicity of the water in water bodies.

PAGE 14
  This again can disrupt the lives of numerous organisms, not only
Artemia.
 One of the best way to put an end to pollution is passing strict
laws, making it compulsory for factories to filter their waste
discharge into water bodies, or completely prohibit it. The same
laws can also be implemented for exhaust smoke.
 PRECAUTIONS:
o Do not use tap water or chlorinated water
o Wash pipette after each use
o Aerate each container at least twice a day for 2 minutes to ensure optimal
oxygen supply
o Place the tanks in distilled sunlight.
o Always prepare a fresh yeast solution.
o Use different stirring rods and droppers for all the tanks to prevent mixing
of mediums.

 BIBLIOGRAPHY:
o https://en.wikipedia.org/wiki/Brine_shrimp
o https://www.britannica.com/animal/brine-shrimp
o https://www.thesprucepets.com/growing-out-
brine-shrimp-2924614
o https://static.fishersci.com/cmsassets/downloads/se
gment/ScienceEducation/pdf/CarolinaBiological/B
rine-Shrimp-CareSheet.pdf

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THANK
YOU

PAGE 16
fin.

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