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 Welcome to our Watershed Project 2 00 8 

Our project required us to study the levels of Coliform in two water sites. This was a year long study; we evaluated the levels of change during different seasons. As a group we went to the Flat Rock Brook Nature Sanctuary in Englewood three times over the course of this year. We collected data from three different seasons to compare the levels of Coliform in the water.Once we collected the water samples from the sanctuary, we put them in petri dishes. We saved multiple samples of the water and stored them in the incubator. This way the water would dry and we could analyze the bacterium left over. The bacteria grouped in red clutters and formed in a honey comb shapes, which was a Coliform colony. 
 * __Objective:__ **

__**What is Coliform?**__ Coliform is a bacterial organism that can be found in soil or vegetation. This organism can also be found in the intestinal tracts of warm- blooded animals. Traces of this organism are a sign of contamination in the water supply. Coliform are easy to identify because they are often present in large numbers and also form big colonies.

Most bacteria need a food source and warm moist conditions for maximum survival. Coliform bacteria are most likely to survive in deep, slow-moving, heavily polluted waters that lack dissolved oxygen. The survival of Coliform bacteria depends on the characteristics of the soil. Survival of coliform bacteria is less likely in well-aerated soil.
 * __How do Coliform bacteria survive?__**

Coliform can be located in various habitats. For example, they can exist in runoff from woodlands, pastures and feedlots, septic tanks and sewage plants, and animal and wild fowl. Domestic animals can contribute heavily to the bacterium population. Many coliform bacteria enter natural streams by the direct disposal of waste into the water system and by runoff from areas with high concentration of humans and animals.
 * __Where are they found?__**

__**Will Coliform bacteria make us sick?**__ Coliform bacteria are not just one type of bacteria; they are a compilation of various types of bacteria. Each bacterium’s specific role defines its purpose. Most coliform are harmless residents of soil and will not make people sick. However, some strains, such as E.coli, the most common fecal coliform bacteria that usually live in animal fecal material, may be pathogens. Some found in food sources have been lethal. For these reasons, their presence should be taken very seriously.

- Filtration - Chlorination - Ultra violet irradiation - Ozonation - Silver treatment - Iodination - Pasteurization By testing water samples for coliform bacteria, one is able to determine the quality of water. The water can be tested for coliform bacteria and fecal coliform bacteria, and these factors indicate the quality of a water source. Fecal Coliform is usually found in the  fecal material from animals; therefore, their presence in the water shows that the water is contaminated by sewage. Finding the <span style="color: rgb(28, 3, 3); font-family: Georgia,serif"> source of the problem and correcting it is very important for the publics safety.
 * __What types of water treatment will make the water safe for consumption?__**
 * <span style="color: rgb(28, 24, 3); font-family: Georgia,serif">__Why test for coliform bacteria?__ **

__**<span style="color: rgb(28, 3, 3); font-family: Georgia,serif">What is the difference between Chloroform and Coliform? **__ <span style="font-family: Georgia,serif">Chloroform is a form or chlorinated organic chemical. It is a volatile organic compound, and coliform describes a group of bacteria that grow on certain types of growth media. It can cause a color change when incubated that tells the analysis that these bacteria are present.<span style="font-size: 150%; color: rgb(0, 0, 0); font-family: Georgia,serif"> <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif">For our watershed project, we decided to analyze coliform levels in two water sources from Flat Rock Nature Sanctuary. One water source was located at the entrance of the sanctuary on the ground level, near the children’s playground. The second water source was located at the top of the sanctuary near houses; however, this second source was more isolated and enclosed in the trees with many rocks surrounding it. <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif"> <span style="font-size: 120%; font-family: Georgia,serif"> __(__ __This is the water source located at the entrance of the sanctuary on the ground level)__
 * <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif">__Introduction:__ **

__**<span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif">Hypothesis: **__ <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif">We think that the coliform levels will be higher during the warmer seasons because bacteria need warm and moist conditions in <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif"> order to grow and survive in a certain enviorment. We predict that the bacteria levels will be higher during the spring as opposed to the fall or winter. <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif"> <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif">- 3m Petra film <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif">- Aerobic count plates <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif">- Petrifilm spreader <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif">- Barrel pipettes <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif">- Incubator <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif">- Plastic containers <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif">- Microscope <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif">- Bio Facts 3M Petra film Aerobic count plates <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif">  <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif">  <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif">
 * <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif">__Material:__ **

<span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif">


 * <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif">

__Method:__ ** <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif">__1__) Place the Petrifilm aerobic count plate on a flat surface <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif">2) Carefully peel open the Petrifilm plate <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif">3) Being careful not to touch the nutrient gel with your fingers <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif">4) With a pipette- perpendicular to the Petrifilm plate, place one milliliter of water from a selected site at Flat Rock Brock on to the center of the bottom film <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif">5) Release the top film; allow it to drop <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif">6) Do not roll the top film down <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif">7) With ridge side down, place the spreader on the top film over the sample <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif">8) Gently apply pressure on the center of the spreader to distribute the water over a circular area <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif">9) Do not twist or slide the spreader, simply apply a gentle, downward pressure <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif">10) Put the Petrifilm plate into the incubator for 24-48 hours <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif">11) After the selective time period is over, retrieve the samples from the incubator and observe <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"><span style="color: rgb(28, 3, 3); font-family: Georgia,serif">12) Now place the Petrifilm plate under a microscope and observe the coliform. <span style="color: rgb(196, 186, 186); font-family: Georgia,serif"> <span style="color: rgb(0, 14, 255); font-family: Georgia,serif"> <span style="font-family: Georgia,serif"> <span style="font-family: Georgia,serif"> <span style="color: rgb(186, 0, 255)"> __

<span style="font-size: 140%; color: rgb(52, 40, 235); font-family: Georgia,serif"> **<span style="font-size: 110%; font-family: Georgia,serif">Data: **__

__<span style="font-size: 120%; font-family: Georgia,serif"> ( [|http://www.m-techdiagnostics.ltd.uk/pics/ecc.jpg]   __


 * [[image:tablewatershed.jpg]] || === === ||

<span style="font-family: Georgia,serif">
<span style="font-family: Georgia,serif">^ The data above shows how many Coliform colonies were apparents in our 1 mm water samples from both the top water source and the bottom water source.

<span style="font-size: 140%; color: rgb(0, 0, 0)"><span style="font-size: 140%; font-family: Georgia,serif"> <span style="font-family: Georgia,serif">Graphs:






<span style="font-size: 120%; font-family: Georgia,serif">**Location:** <span style="font-size: 140%; font-family: Georgia,serif"> <span style="font-family: Georgia,serif"><http://www.flatrockbrook.org/features/trails.html>

From the data presented in the graphs, it is clear that our hypothesis was accurate. The Coliform levels were higher during the spring since the water and surrounding temperatures were higher. The Coliform levels were lower during the cooler months in the fall and winter. From our data, we also discovered that the levels of Coliform are higher in the lower water source compared to the higher water source. The reason for this is because the lower water source gathers all of the bacteria that travels through the nature sanctuary and collects in the bottom water source.
 * __Analysis:__**

Ultimately, after collecting and analyzing our data, we have found that our hypothesis is accurate. The Coliform levels were higher during the warmer months and lower during the cooler months. This result makes sense because coliform needs warm moist conditions to grow successfully. We also found out that the levels of coliform were higher in the lower water source (on the ground level of the sanctuary) compared to the top water source, regardless of the season. The reason for this is the lower water source was still water and the top water source was running water. This means that the water from the top water source flows down collecting various types of organisms and collects in the lower source. From this point on, (in the lower water source) the water collects and the coliform is able to grow in the still water.
 * __Conclusion:__**

<span style="font-size: 110%; font-family: Georgia,serif">__**Possible Experimental errors:**__ During the whole process, our team could have made several experimental errors due to the many variables involved in our research: 1) Temperature: As the seasons change the temperature of the water changes. We did not measure the temperature of the water and this factor could have had an impact on our results. 2) Location: We might not have taken the water samples from exactly the same location on the two water banks. Different water banks could collect different amount of bacteria depending on its surroundings; for example, various sources of rocks, vegetation and animal exposure. 3) Incubation Time: The samples were not in the incubator for the same amount of time. They varied from 24 hours to 48 hours. 4) Air bubbles: It was very had to get the air bubbles out of the Petri Dish, which could have affected the growth of Coliform. 5) Time: Every time we went to Flat Rock Brook, we went at different times of the day, and linked with the air temperature on the given days, this could have effected the levels of coliform in the water. <span style="font-size: 110%; font-family: Georgia,serif">6) The type of weather (rain, snow...etc) could have effected the water bodies and the amount of sewage that might have seeped into the water.

<span style="font-size: 80%; font-family: Georgia,serif">** Sources: **http://www.m-techdiagnostics.ltd.uk/pics/ecc.jpg http://www.flatrockbrook.org/features/trails.html http://www.wellowner.org/awaterquality/coliform.shtml <span style="font-size: 80%; font-family: Georgia,serif">

<span style="font-size: 80%; font-family: Georgia,serif"> By: <span style="color: rgb(86, 30, 179)">Isabella Rodrigues, <span style="color: rgb(247, 0, 255)">Gabrielle Melton, <span style="color: rgb(179, 20, 172)">Megan Kothari