EFFECT OF TEMPERATURE ON HATCHING SUCCESS

The Effect of Temperature on the Hatching Success of Brine Shrimp Aim To investigate the effect of temperature on the hatching success of brine shrimps. Introduction Brine shrimps (Artemia salina) are small Crustaceans that populates in the extreme environment of salt lakes. Brine shrimps feed on a type of phytoplankton that is suspended in the water. The lack of competitors or predators in the extreme environment of salt lakes allows these brine shrimps to reproduce in large populations during seasons when the conditions for reproduction are suitable. Factors that affect reproduction of the brine shrimps are temperature, sunlight, salinity, and presence of water. The life cycle of a brine shrimp is only 12 months and usually begins in early spring. After hatching, the larvae will go through 15 molting cycles before it reaches the reproductively mature adult form. These adults can reproduce live young in rapid succession in favorable conditions, and also by eggs cysts that can endure prolonged periods of dryness in unfavorable conditions. The larvae will emerge from the egg cysts when the conditions for development and reproduction are favorable again. These egg cysts will be scattered by winds and waves, and are often brought to shore, where they remain dormant until rain falls in spring and washes them back into the sea. Dry eggs cysts are between 0.2 0.3 mm in diameter and it will hatch in seawater into free-swimming larvae of about 0.45 mm in a period of 2436 hours. In the absence of males, female brine shrimp are able to reproduce by fertilizing their own eggs, a process known as parthenogenesis. The eggs produced in this method typically have thinner shells those that are sexually produced and are usually female. In this experiment, the effects of temperature on the hatching success of the brine shrimps are investigated. Hypothesis As the temperature increases, the number of successfully hatched brine shrimps will increase too. The optimum temperature for the brine shrimps to hatch successfully is between 20-30.

EFFECT OF TEMPERATURE ON HATCHING SUCCESS

Material 1. Salt solution 2. Beakers 3. Brine shrimp egg cysts 4. Filter paper 5. Graph paper 6. Magnifying glass 7. Scissors 8. Forceps 9. Incubators at temperatures of 5C, 20C, 30C, and 35C 10. Bench lamp Methods 1. Eight beakers of salt solution were prepared beforehand and two beakers each were labeled with temperature of 5C, 20C, 30C, and 35C. 2. A tiny pinch of egg cysts were placed onto a sheet of filter paper. 3. A piece of graph paper was wetted using a few drops of salt water. The paper was dabbed onto the filter paper to pick up approximately 40 eggs. A magnifying glass was used to count the eggs. The graph paper was cut so that there are exactly 40 eggs. 4. The paper with 40 eggs was put into one of the beakers with the eggs-side facing down. After 3 minutes, the paper was gently removed using a pair of forceps. The egg cysts were made sure to have all washed off into the water. 5. Steps 2 to 4 were repeated for all the temperatures to be investigated. The beakers were incubated at the appropriate temperatures. 6. Two days later, the number of hatched larvae in each beaker is counted by placing a bright light next to the beaker. The larvae will swim towards the light. 7. The number of larvae that have successfully hatched at each temperature is recorded.

EFFECT OF TEMPERATURE ON HATCHING SUCCESS

Results Table 1: Number and percentage of brine shrimps hatched in different temperature Temperature (C) 5 20 30 35 Beaker #1 0 6 10 1 Beaker #2 0 5 4 2 Average 0 5.5 7 1.5 Percentage (%) 0 13.75 17.5 3.75

Graph of Percentage of Brine Shrimps Hatched (%) vs. Temperature (C)

Percentage of Brine Shrimps Hatched, %
20 18 16 14 12 10 8 6 4 2 0 17.5 13.75

3.75 0 5 20 30 35

Temperature, C

Discussion The percentage of the brine shrimps hatched is calculated using the formula

The graph shows the relationship between the temperature and percentage of eggs hatched. According to the graph, as the temperature increases, the percentage of brine shrimps hatched increases as well, until it reaches 30C, which is the optimum temperature. At temperatures beyond 30C, the hatching percentage of brine shrimps decreases significantly. In this experiment, the salinity and pH of the salt solution are controlled as they are both factors that can affect the hatching success of the brine shrimps. To keep the salinity constant, the same amount of sea salt is used in each beaker of water. Having less salt in the

EFFECT OF TEMPERATURE ON HATCHING SUCCESS

water makes it easier for the brine shrimp to absorb enough water in order to burst through the protective shell of the cyst; while having more salt will consume more of the energy reserves of the embryo. This is because when less salt is used, the osmotic pressure of the water surrounding the egg cyst is higher compared to the internal condition of the egg, making it easier for water to go into the egg cysts. Optimal salinity for hatching varies from strain to strain, but is generally situated in the range of 15-70 g.l-1. The pH of the salt solution is maintained between 8.0 and 8.5 because it is close to the natural pH of the salt lakes, which is slightly alkaline. If pH is below 8.0, the hatching rate will decrease significantly (Lee, 2011). The hatching of the brine shrimp egg cysts requires a certain enzyme known as the hatching enzyme (HE). HE is secreted from hatching gland cells of the hatching larvae and functions in the digestion of their protective extracellular coats. The enzyme affects hatching by activating the diapausing cysts and the quiescent portion in a suitable environmental (Li, Fan, Wang, Cong, Yu, & Zhong, 2004). As the hatching of egg cysts depends on an enzyme, it explains why there was no egg cysts hatched at the low temperature of 5C. At such low temperature, the hatching enzyme is inhibited. As the temperature increases, typical of all enzymes, the enzyme reaction will increase too. This leads to an increase in the hatching of the brine shrimps egg cysts. Most egg cysts are hatched at 30C; therefore it is the optimum temperature for the hatching success of brine shrimps. Beyond the optimum temperature of 30C, the enzyme reaction decreases and comes to a complete stop at temperatures after 40 C. This is because the hatching enzyme has already been denatured by the high temperature. A few egg cysts are hatched at 35C because the enzyme is not fully denatured yet. The brine shrimp larvae swim towards the light due to phototaxis. This is because the phytoplankton the brine shrimp feeds on are photosynthetic, therefore these phytoplankton tend to be more abundant at the surface of the water and where there is more light. Positive phototaxis in the brine shrimp larvae helps it to obtain food (Lee, 2011). Conclusion As the temperature increases, number of brine shrimps hatched until it reaches the optimum temperature. The optimum temperature for the brine shrimps to hatch successfully is 30C.

EFFECT OF TEMPERATURE ON HATCHING SUCCESS

Reference Lee, N. Y. S. (2011, February 2). Practical 5.2: the effect of temperature on the hatching success of brine shrimps [Web log]. Retrieved from http://brainstormthoughts147.blogspot.com/2011/03/practical-52-effect-oftemperature-on.html Hickman, O. (2006, March 5). Brine shrimp [PDF document]. Retrieved from http://fog.ccsf.cc.ca.us/~cpogge/Bio%2025/BrineShrimp.pdf Li, L., Fan, T. J., Wang, X. F., Cong, R. S., Yu, Q. T., & Zhong, Q. W. (2004). Immunocytochemical studies on the phase of differentiation of hatching gland cells in brine shrimp, Artemia salina [Abstract]. Shi Yan Sheng Wu Xue Bao, 37(2), 157164. Retrieved from http://www.ncbi.nlm.nih.gov/pubmed/15259990