Brine Shrimps

UNIVERSITI TEKNOLOGI MARA INTERNATIONAL EDUCATION CENTRE INTEC 2011
Name IC.NO. Group
: Subbashini a/p Sekar Chandran : 921129-14-6112 : 11M11
Student ID : 2010699896 Experiment No.: Practical 5.2 Title : The Effect of Temperature On The Hatching Success Of Brine Shrimps Date Lecturer : 8th March 2011 : Dr.Noraini
Subbashini Sekar Chandran (11M11) Biology Lab Report- The effect of temperature on the hatching success of brine shrimps
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ABSTRACT The purpose of this experiment is to investigate the effect of temperature on the hatching success of brine shrimp. The experiment was carried out at different temperatures to see the effects of it on the brine shrimp egg cysts. 2 different temperatures were used, these were room temperature and 35C . The number of the shrimps hatched at each temperature was counted by viewing a sample under a binocular microscope. This process was repeated twice for each temperature in order to get the mean number of shrimps for each temperature. The percentage of shrimps hatched for each temperature was also calculated. The number of hatched eggs at room temperature which was 28C was the highest, followed by 35C which had the least number of hatched shrimps.
INTRODUCTION Brine shrimp is the English name of the genus called Artemia of aquatic crustaceans. Artemia is the only genus in the family Artemiidae and have evolved very little since the Trassic period. The historical record of the existence of Artemia dates back tomore than a thousand years ago. Artemia are found worldwide in inland saltwater lakes but not in oceans. The life cycle of brine shrimp starts from an egg. Brine shrimp eggs are metabolically inactive and can remain in complete stasis for 2 years while in conditions which are dry and oxygen free. They can even withstand temperatures below freezing point. This is called cryptobiosis. Brine shrimp can also survive temperatures of liquid air and some even can survive boiling temperature. Once the eggs are placed in salt water, the eggs will hatch within few hours. The larvae are shorter than 0.5mm in length when they first hatch. They have a life cycle of one year. For the first few hours the embryo hangs beneath the shell, still enclosed in the hatching membrane. The larvae do not feed in the first few hours after they are born because their mouth and anus have not fully developed. Once they start feeding, they feed on microalgae, bacteria and many others. Word Count: 337
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The larvae will reach adulthood in about 8 days by time they would have undergone about 15 moults. Brine shrimp can grow to about a length of 1cm but can reach to lengths of about 2cm in the right environment with the right conditions. Adult brine shrimps can tolerate brief exposures to extreme temperatures. They also prefer salinity of about 30-35 ppt and can live in fresh water for about only 5 hours before they die. pH, light and oxygen are also very important factors that can affect the survival of these organisms. pH of 8-9 is the best of these organisms. Only a minimum amount of light is needed for their growth. The concentration of oxygen in the water is the most important because this determines what the brine shrimps will consume. With good concentrations of oxygen, the brine shrimp are pale pink or yellow. Sometimes they may be green because they feed on microalgae. Reproduction of these organisms are rapid in conditions like this. Under different conditions, the brine shrimp may feed on bacteria and yeast cells but not algae. It is under conditions like this that they produce haemoglobin and look red in colour, sometimes even orange. They will start reproducing if the environmental conditions remain like this. They produce resting cysts.
Brine shrimps
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The stereo microscope is designed to serve 2 different purposes. It separates optical paths with 2 objectives and 2 eyepieces to provide different viewing angles to the left and right eyes. It produces a 3D visualization of the sample being examined. It is used to study the surfaces of solid specimens and to carry out close work. It is capable of transmitted and also reflected illumination. Reflected illumination enables objects that would normally seem opaque to be seen when compound microscopes cannot view them as light cannot pass through. This type of microscope has a great working distance and the depth of field is also an important quality of this microscope. These qualities are inversely correlated with resolution. There are 2 types of magnification for this microscope, which is fixed magnification and pancreatic magnification. A stereo microscope provides greater viewing comfort but the image in such a microscope is no different from that obtained with a single monocular eyepiece.
An optical microscope
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EXPERIMENTAL HYPOTHESIS The higher the temperature, the lower is the hatching success of brine shrimps NULL HYPOTHESIS The hatching success of brine shrimps is independent of changes in temperature.
VARIABLES Manipulated variable : Temperatures of saline solution ( room temperature and 35C) Responding variable : Number of hatched larvae for each temperature Control variable : Amount of eggs used, concentration of sodium chloride solution
MATERIALS AND APPARATUS Brine shrimp egg cysts, 1% sodium chloride solution, binocular microscope, boiling tubes, spatula, water bath of temperature 35C, Petri dishes, measuring cylinder, a piece of graph paper, bright light, magnifying glass, stirring rod and droppers.
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METHOD 1. A tiny pinch of egg cysts was placed onto a large sheet of white paper. 2. A small piece of graph paper was wet using the salt water prepared beforehand. The paper was dabbed onto the white sheet to pick up approximately 40 eggs. This looked like a tiny shake of pepper grains. 3. A magnifying glass was used to count the eggs and the graph paper was cut so that there were exactly 40 eggs. 4. The paper with the 40 eggs was put into a boiling tube containing 25 cm of 1% sodium chloride solution (egg-side down). After 3 minutes, a pair of forceps was used to gently remove the paper, making sure that all the egg cysts were washed off into the water. 5. Steps 1 to 4 was repeated for both the temperatures which were to be tested and both treatments had two replicates. 6. Two of the boiling tubes were placed in a test tube rack at room temperature while the other two set-ups were incubated in a water bath at 35C. 7. The next day, the number of hatched larvae in each boiling tube was counted. To do this, a bright light was placed next to the boiling tube. The hatched larvae swam towards the light. The brine shrimps were then caught using a dropper and were then placed in a Petri Dish. 8. The brine shrimps were also counted under the stereo microscope to reaffirm our counting. 9. The countings were repeated for three days. Since the brine shrimps were very delicate, care was taken when handling them. 10. The number oflarvae hatched in each replicate for each type of treatment was measured and the mean number was calculated. 11. The percentage of hatched shrimps were calculated by using the following formula:
12. The data obtained were tabulated in a table and graphs were plotted using these data.
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RESULTS TABLE 1 (Raw Data) NO. OF HATCHED BRINE SHRIMP TEMPERATURE Room Temperature (28C) Test tube DAY 1ST 2ND 3RD I 14 9 3 II 11 8 2 I 2 2 1 II 0 1 1 35 C
TABLE 2 (Average values of hatched larvae) TEMPERATURE NO. OF HATCHED SHRIMPS
ROOM TEMP ( 28C) DAY 1 2 3 12.5 8.5 2.5
35C
1 1.5 1
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TABLE 3 ( Percentage of hatched larvae) TEMPERATURE DAY 1 2 3 PERCENTAGE OF HATCHED SHRIMPS (%) ROOM TEMP (28C) 31.25 21.25 6.25 35C 2.5 3.75 2.5
Graph 1
graph of number of shrimp eggs hatched against the number of days for two temperatures
14 12 10 Number of hatched 8 eggs 6 4 2 0 0 1 2 Days 3 4
28C 35C
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Graph 2
Graph of percentage of hatched brine shrimps for three consecutive days at two different temperatures
35 30 25 Percentage of 20 hatched brine shrimps (%) 15 10 5 0 0 0.5 1 1.5 Days 2 2.5 3 3.5 28 35
DISCUSSION This experiment was carried out to investigate the effects of temperature on the hatching success of brine shrimp egg cysts. The experiment was carried out at different temperatures too see how it would affect the number of eggs hatched. The experiment was carried out at room temperature which was 28C, and also 35C. As the results have shown, the most number of eggs hatched at 28C which is 31.25 on the first day in 25ml of NaCl solution. This is followed by 35C with approximately only one egg hatched on the first and third day. There were a very little eggs hatched at 35C because the enzymes involved in the hatching of the brine shrimp have been denatured at this high temperature. This is because the bonds holding the enzyme together and maintaining its shape are broken down as the temperature increases The absence of enzymes interferes with the hatching process of these organisms Furthermore, the embryos in the eggs would also be killed at this higher temperature. That is the main reason why there is such a drastic decrease in the percentage of
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eggs hatched as the temperature increases. For instance, on Day 1, 31.25% of eggs in the boiling tubes at room temperature had hatched while only 2.5% of their counterparts in the boiling tubes at 35C had hatched on that same day. Generally, both graphs indicate that the success rate of brine shrimp hatching is about 12.5 times higher at a lower temperature, which is room temperature, than at a higher temperature, which is 35C. When doing this experiment in particular we have to look at some issues that are ethical and by this I mean when testing shrimps for the use of laboratory experiments. I contend to the use of eggs for the sake of science as it is a way for us to obtain a better understanding of the world we live in. We also released the brine shrimps after use back into the water once we were finished with them. I believe that the ethical issue will be brought up when we recklessly use a high number of shrimp egg cysts than the amount actually needed. The manipulated variable in this experiment is the temperature of the environment the eggs were hatched in. The temperatures were manipulated by placing the boiling tubes in a test tube rack at room temperature and water bath at 35C. This enables us to observe the effects of different temperature on the success of hatching of brine shrimp. The responding variable in this experiment is the number of eggs hatched. This is found out by counting the shrimps swimming about while viewing it under a stereo microscope. Due to the moving crustaceans, it is likely that the first data obtained is not accurate. So a second reading is taken by different people and the mean is calculated. This average will provide more reliable data. This is done for both the temperatures to get a more accurate result. There are also some factors which were kept constant throughout the whole experiment. The amount of eggs used was kept constant in this experiment. This is to make sure that the number of eggs hatched is not due to the different amount of eggs present in each boiling tube but only due to the effects
Subbashini Sekar Chandran (11M11) Biology Lab Report- The effect of temperature on the hatching success of brine shrimps Page 10
of temperature. The salinity or concentration of sodium chloride is also kept constant because this is one of the factors that affect the hatching of brine shrimp.
CONCLUSION The hatching success of brine shrimp is affected by temperature whereby the higher the temperature, the lower is the success rate. The hypothesis is accepted.
LIMITATIONS AND SOURCES OF ERRORS Limitations There is one limitation in this experiment. That is whether to count the hatching brine shrimps that are just coming out of the cysts as hatched or not hatched. This might happen if the sodium chloride wasnt left long enough to allow the eggs to hatch. Furthermore, the hatching eggs or hatched eggs arent obvious under the stereo microscope. This might lead to a different count of brine shrimp each time a count is done and this might make the results less reliable.
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Sources of Errors There are a few sources of error in this experiment. The first one would be the number of eggs used for each experiment. There is an error because we counted the eggs which were scattered on a piece of graph paper with just the aid of a magnifying glass. The egg cysts of these organisms are like pepper grains and to make matters worse, some of these eggs are clustered and are hard to count even with a microscope. There might be a different number of eggs because the amount is solely based on individual judgement. This might lead to less reliable results. Another source of error is when calculating the total number of eggs and also the shrimps hatched in each boiling tube. This is because the shrimps are moving creatures and so, incidence of counting the same shrimp more than once is highly likely. Another error is that we do not know that the room temperature is constant and when left over a couple of days it is of no surprise to find that the temperature at night may not necessarily be the same as it was in the day.
FURTHER WORK An extension of this experiment can be done by changing the salinity of the sodium chloride instead of carrying out the experiment using different temperatures. This is because the salinity is also one of the important factors that affect the hatching of brine shrimp eggs. The set-up of the experiment will be similar, but instead of different water baths (the temperature is kept constant), different concentrations of salt solutions are used and the number of eggs hatched are observed. The number of brine shrimp hatched will be higher in low saline concentration and lower in high salinity as the high salinity might kill the embryos. Word Count: 302
BIBLIOGRAPHY
1. http://en.wikipedia.org/wiki/Brine_shrimp
Date acquired: 10th March 2011 2. http://web.cecs.pdx.edu/~davidr/discus/articles/artemia.html Date acquired: 10th March 2011 3. http://en.wikipedia.org/wiki/Optical_microscope Date acquired: 10th March 2011
THE END
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UNIVERSITI TEKNOLOGI MARA INTERNATIONAL EDUCATION CENTRE INTEC 2011

Name IC.NO. Group

: Subbashini a/p Sekar Chandran : 921129-14-6112 : 11M11

Word Count: 217

Word Count: 162

Word Count: 328

TABLE 2 (Average values of hatched larvae) TEMPERATURE NO. OF HATCHED SHRIMPS

ROOM TEMP ( 28C) DAY 1 2 3 12.5 8.5 2.5

Word Count for page 9, 10 and 11: 698

Total Word Count : 2,341 words

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