The most harmful pollutants are the industrial wastes and byproducts because they are sometimes overlooked. Factories are often located near bodies of water where they dump all their wastes Into. The metallic wastes that come from Industrial factories are very harmful to marine life In a way that It can either cause mutations or moralities. The threat of heavy metal poisoning on organisms In the aquatic ecosystem often seems to be Ignored; Nothing the surrounding ecosystems. The intended audiences for this thesis are the locals.
Lead acetate, primarily used in dyeing and printing textiles, drying paint and varnishes, and can be found in eateries, pesticides and gasoline, is accessible to anyone and therefore can be misused by anyone if they are not informed of its effects to the environment. It is a lethal toxin and can cause behavioral, neurological and cognitive defects. Trustiness guerilla is a commercially valuable tropical sea urchin with high growth rates. It is being targeted as an aquaculture species in several countries, is produced in small quantities for restocking in Japan, and as food in the Philippines.
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This project focuses on how pollution affects the aquatic ecosystem, simply because he Earth is mostly made of water, which is very essential to life. The embryonic development of every organism determines its biological success later in life. The thesis would explore the effects of lead acetate in the embryonic development of sea urchins. Sea urchins are common and quick in reproducing, making the thesis less damaging to the food web. In the experiment, the sea urchins (Trustiness guerilla), were injected with KICK for them to spawn.
After collecting the gametes of the sea urchins, they will be tested to develop under 4 different set ups. The first containing 5 pump lead acetate incarceration, the second with 10 pump, the third with 15 pump, and the last with the normal environment as the negative control. The embryos were examined under the microscope after 2 hours and the number of unfertile eggs, delayed embryonic development; successful 4-cell embryonic stage and deformed embryos were tallied and recorded carefully.
After conducting the experiment, data shows that the higher the concentration of lead acetate the gametes were exposed to, the less likely the eggs were to have successful embryonic development. The percentage of unfertile eggs, placement delay and deformed embryos increased as the concentration of lead acetate increased. However, as the lead acetate concentration got higher, the percent average of successful embryos that undergone 4-cell stage decreased. This only shows that truly, lead acetate as an aquatic pollutant, affects the growth and development of the tested Trustiness guerilla.
REVIEW OF RELATED LITERATURE ‘One of the greatest threats to the health of the environment today is the issue of pollution, particularly in aquatic ecosystems where pollutants can enter the system kilometers away and accumulate from multiple sources. Fertilizers, pesticides, industrial pollutants and waste byproducts are frequently discussed in terms of Neater pollution and their effects on the ecosystem. Of these pollutants, industrial Negates and their heavy metal byproducts are often overlooked from an environmental standpoint.
When industries are located in areas near agricultural or recreational areas heavy metal pollution is considered a potential threat from the industry’ (Radiance, Fiches, Miranda 2001). The threat of heavy metal poisoning on organisms in the ecosystem often seems to be overlooked; however, many of the active effects of heavy metals on humans apply to organisms within the nickel, can all act as poisons or teratology, causing deformities or deaths in many organisms exposed to them. Organisms in aquatic systems, particularly smaller organisms, are often subject to heavy metal poisoning.
When the metal is dissolved onto the water column it becomes a significant aquatic contaminant that affects the organisms living within the ecosystem. Heavy metals can also settle in large quantities along river or coastal beds’ sediment (Armenia, Fiches, Miranda 2001). These heavy metals can be one of the most destructive anthropogenic pollutants. The effect of heavy metals on early development, particularly on early embryonic stages, can be detrimental to a species that occupies a habitat into which heavy metals are introduced.
Aquatic pollution has been a source of biotic imbalance in ecosystems. It is caused by different impurities and chemicals released to different bodies of water, n particular, oceans and seas. One of these metallic impurities underwater is Lead, in particular, lead acetate. Lead (II) Acetate (BP(CHICHI)2), also known as Lead Acetate, is a white crystalline chemical compound used as a mordant in textile ranting and dyeing and as a drier in paints and varnishes. Like all other Lead compounds, Lead Acetate is a highly toxic substance.
Lead interferes with the proteins that cause certain genes to turn on and off by displacing other metals in the molecules. It can displace other nutrients of developing organism, causing more harm than nutrition. Lead can enter water systems through runoff, sewage, and Industrial waste systems. When industries are located in areas near agricultural or recreational areas, heavy metal pollution is considered a potential threat to marine organisms. (Radiance, Fiches and Miranda , 2001).
Both these natural and unnatural processes have resulted to an increase in the amount of Lead and more importantly, the disruption of the biochemical cycle. Because of its high toxicity, this metal has caused reduced growth, reproductive malfunction and reduced survival of organisms. These effects were evident in the organisms’ life stages. One organism that is easy to observe in development is the sea urchin, which lives in marine environments that are often subjected to heavy metal pollution, particularly in regions where there are coastal industries.
Sea urchins are particularly susceptible o environmental conditions; in fact sea urchins exposed to toxic conditions have Inhibited embryonic development and growth (Quinoa, Guilty, Judas 1999). The fact that sea urchins respond so readily to environmental conditions makes them an ideal species to act as an indicator of ecosystem health. They are also one of the best vindicators to determine heavy metal pollution because of the variety of developmental anomalies that are caused by the different heavy metals (Sickbay’s and Kumara, 2004).
Most Echinoderms, Escondido or Sea Urchins in particular, are the most favored organisms for study in the field of development biology. Their development is easily studied because they release a massive number of gametes Nee spawned. Other reasons are that sea urchins exhibit cell transparency and a rapid rate of cell division. Location wise, these organisms live in environments that are constantly subject to heavy metal pollution. Because of these, sea urchins have become one of the best vindicators to determine heavy metal pollution because of the variety of developmental anomalies caused by different heavy metals. Sickbay’s and Kumara, 2004) Trustiness guerilla Embryonic Development Hours After Fertilization 150 2 cell 1:10 :55 4 cell 8 cell 3:30 2:15 16 cell 12:00 4:30 Early blastula 43:o 10:o Early gastrula 25:o 18:00 Prism 26:o Pluses (Wilson and Hey, 1970) RATIONALE Trustiness guerilla is a commercially valuable tropical sea urchin with high growth rates (Dormancy et al. , 2007; Lawrence & Satsuma, 2001). It is being targeted as an aquaculture species in several countries, including Australia (Dormancy et al. , 2007; loin-Mines et al. , 2008; Moms et al. 2011; Simulators, 1991), is produced in small food in the Philippines Nonunion-Mines et al. , 2008). The intended audiences for this thesis are the locals. Lead acetate, primarily used in dyeing and printing textiles, drying paint and varnishes, in batteries, pesticides and gasoline, is accessible to anyone and can be misused if some are not informed of its effects to animals. It is easily spread to organisms in air, water and food. It is a lethal toxin and can cause behavioral, neurological and cognitive defects. Findings in this thesis could be compared and applied to other organisms.
It Mould explore on where the development goes wrong and how the damage could possibly be avoided. The thesis would not include comparisons of the effects of lead acetate on the embryonic development of other organisms. Different organisms would not always respond to chemical stimuli in exactly the same way. The findings in this thesis may not always be identical with the findings in similar theses using other organisms. Alternative works to be read are those covering the effects of lead acetate in other organisms, the elimination of lead acetate in household products and determining lead acetate content.
OBJECTIVES 1 . Find out the effects of lead acetate in the sea urchins embryonic development specifically during 4 cell stage. 2. Make an inventory of the observations found in ACH sample. 3. Determine the mean percentages of embryonic developmental delay, abnormalities, mutations or unfertile eggs should they occur. SCOPE AND LIMITATIONS The scope of this study is limited on the application of lead acetate in the sea urchins (Trustiness guerilla) embryos until the 4-cell stage embryonic development.
METHODOLOGY Preparation of Sea Urchin Embryos and Test Solutions ripeness’s guerilla adults were collected at Cordovan, Lap Lap, Cube City during the month of September 2013, and were maintained in the laboratory in aerated sea Neater at room temperature (220). Three concentrations of lead acetate with 5 pump, 10 pump and 15 pump were created to test its effects on the sea urchins embryonic development, specifically during 4 cell stage. These concentrations were tested thrice, along with a sample in sea water to Using a syringe, the researchers injected 1-2 ml of KICK (0. 3 M = 3. 9%) in the peristalsis membrane near the Aristotle lantern for the spawning. After 2-5 minutes, the process was repeated. As soon as the spawning begun, the researchers checked the color of the gametes to identify its gender (sperm are creamy white; eggs are yellow, pink or dark red). For the male sea urchins, the first sperm was Immediately transferred in the Petri dish to get rid of perspectives fluid that would interfere with the sperm’s ability to fertilize and allowed the urchin to shed into the Petri dish without diluting the sperm.
For the females, the eggs were shed into the seawater by placing her, inverted, on top of a 100 ml beaker. The beaker was full enough that the seawater touches the abroad side of the urchin where the eggs were drifted down through the seawater and settled in the bottom of the beaker. Two drops of dry sperm were suspended in 10 ml of seawater then a drop of this suspension was poured to a plastic container (C=14. 14 inches) which contains another 10 ml of seawater containing the eggs. The container was then mixed, in a circular motion, by Just holding the container.
The same process was repeated using the three other set-ups each contained 5 pump, 10 pump, and 15 pump of lead acetate concentrations. After two hours, one drop sample from each set ups were observed under the microscope (1 Ox). Three different samples from each set-up were observed for more accuracy. The results were carefully observed and recorded for the results and discussions. RESULTS AND DISCUSSIONS Table 1 shows the percent of unfertile egg observed in the study. According to the table, 1 1. 4% of the total embryos exposed under the normal environment were unfertile. While embryos under the 5 pump, 10 pump and 15 pump lead acetate concentrations were 24. 61%, 55. 12% and 60. 30% unfertile respectively. Table 2 shows the percent of developmental delay observed. Embryos undergoing 2-cell stage were under the developmental delay because they didn’t reach the 4-cell stage within 2 hours which is the expected stage during the duration. According to the data, 16. 1% was undergoing 2-cell stage under the normal environment.
Embryos exposed in 5 pump, 10 pump and 15 pump lead acetate concentrations had 20. 35%, 23. 05% and 23. 31% of their population with delayed development, respectively. Of the total embryos under normal environment successfully reached the 4-cell stage. 44. 04%, 9. 76% and 2. 10% were the percent of successful 4-cell stage embryos exposed in 5 pump, 10 pump and 15 pump lead acetate concentrations respectively. Table 4 shows the percent deformed embryos observed. According to the table, embryos exposed under the normal environment showed no deformations.
Embryos suspended in 5 pump, 10 pump and 15 pump concentrations had 11. 01%, 12. 06% and 14. 29% of their population being deformed. Graph Presentations The higher the concentration of lead acetate the gametes were exposed to, the less likely the eggs were to be fertilized. When the gametes were exposed to 5 pump concentration of the chemical, the percentage of unfertile eggs observed increased. From an average of 11. 64%, it rose to an average of 24. 61%. When the concentration was made 10 pump, the percentage of unfertile eggs observed rose. If the percentage was 24. 1% in a 5 pump concentration, it would not be double this in concentration that is double 5 pump. It rose instead to 55. 12%. The total number of unfertile eggs in two set-ups each having 5 pump would be less than the number unfertile eggs in a single set-up having the same number of eggs the previous 2 set-ups would have combined. When the concentration was made 15 pump, the percentage of unfertile eggs rose from 55. 12% to 60. 30%, even though the same amount of parts per million was added to the previous concentration of 10 pump, which is 5 more parts per million.
It was also found that the percentage of developmental delay observed in sea urchin embryos in 1 5 pump concentration of lead acetate was almost the same as the percentage in the 10 pump concentration. From 23. 31% under 15 pump, it lowered to 23. 05%. From 23. 05%, it again lowered to 20. 35% when the concentration was lowered to 5 pump. From 20. 35%, it lowered to 16. 71% when the embryos were suspended in normal environment. From this, we can say that the number of embryos that hit the 2-cell stage after the 2 hours duration was highest in 15 pump concentration, followed by the embryos under 10 pump and 5 pump.
More developmental delays were observed as the concentration got higher. This concludes that lead acetate had an effect on developmental delay. The percentage of embryos that entered the 4-cell stage lowered as the concentration of lead acetate increased. 71. 65% of embryos entered the stage in the normal environment. This percentage lowered to 44. 04% when 5 pump of the chemical was added. When 5 more parts per million was added to the concentration, the percentage dropped to 9. 76%.
There would be a smaller number of embryos hitting the 4-cell stage in a set-up with a 10 pump concentration of lead acetate than the total number of embryos hitting the stage in 2 set-ups each having a 5 pump incarceration of lead acetate, and half the number of embryos in the first set-up. When the concentration was increased to 15 pump, the percentage of embryos that entered the 4-cell stage further lowered to 2. 10%. In the normal environment, none of the embryos were deformed. Deformation came and increased as the concentration of lead acetate increased.
When the concentration was 5 pump, the percentage of deformed embryos observed was concentration of 5 pump was doubled. When the same amount of parts per million Nas added to 10 pump, which was 5 more parts per million, the increase in percentage Nas higher than Just near 12. 06%. It rose from 12. 06% to 14. 29%. It is recommended for further related researches that a wider range of concentrations of lead acetate be used in order to see if the trend continues with increasing concentrations and to see where in between them the change in lethality becomes significant.