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Mrs. Adams
Chemistry Pre-AP A3
12/02/10
à
Bacteria are microorganisms that linger everywhere on earth. There are some bacteria beneficial
to our health, yet dreadful ones that influence one¶s internal equilibrium systemic functions
negatively. From this fact, my group began investigating analytically into the sciences of
Bacteria itself and its reactions to certain subjections. We hence came across an article called
³Why Is It So Difficult to Eradicate Salmonella?´ (ScienceDaily) Within this article, which our
project is based primarily on, elucidates how salmonella bacterium is derived from the
salmonella on agar plates, concluding in its formation of biofilm. From our readings, biofilm are
usually composed of synthetic materials in forms of slime. This biofilm induces a resistance
coating, making it harder for chemicals to eradicate Salmonella. We all were baffled by the
all started conceiving the fact whether applications are practical in destroying the biofilm. We
came to a result having the notion of subjecting the bacteria to certain conditions. And since
Salmonella was already hard to eradicate, we pondered whether our hypothesis was valid upon
various bacteria including E. coli. We summarized potential occurrences to that the bacteria may
develop an adaption to certain allocations of pH and temperature and become resistant (similar to
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antibiotic resistant bacteria). And at low ph (acidic) and elevated temperatures, E. coli, at first,
may have a potential of growth rate inhibition, but can mutate to their contiguous conditions for
survival and multiply. Another idea we proposed was how the utilization of acidic conditions
within the nutrients may induce deficiencies for the bacteria, such as a deoxydation or
dehydration mechanism. We came to a conclusion that these bacteria would then be void of
oxidation and water, ceasing reproduction at a quicker rate. This general science article was very
:
Upon the conclusion of our science fair project to universally be in regard to microbiology
(bacteria specifically), we have gathered four professionally research articles in relevance to our
general topic. These articles include mainly the bacterial internal functions subjected to
fluctuations, and growth models for bacterial cells. In one article, ³The Combination of Zinc
Marker Strain in Rich and Minimal Media´ (Park, S.Y., et al. 199-207), researchers¶ utilized
Amalgamated compounds of zinc and acidic pH levels which influenced aerobic growth on a
certain bacteria known as Salmonella Typhimurium. With Ph levels below .05 significantly
inhibited or ceased growth of this bacterium. The growth rates of S. Typhimurium significantly
decreased as a combination of poultry isolate was present in zinc acetate than zinc sulfate.
Growth rates of the bacteria itself were also decreased in minimal Medias than in comparisons to
rich Medias. In pertinence to this article mentioned, our group figured to see how an alternate
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bacterium situated within intestines of organism, Escherichia coli, may be influenced chemically
when pressured at certain pH values between .01 and 1 with strong corrosive acids. Another
article we took into consideration was "Growth and genetic responses of Salmonella
gastrointestinal tracts of poultry. This fact and event is hence analyzed to see any potential of
shifts of 6.17 to 7.35 which was critical in recording the outcome at that variable. By doing so,
they observed how increases in pH values of chemicals/ substances, resulted in higher cell
protein concentrations, glucose disappearance, and glucose and ATP yields. At high pH
influences, Salmonella Typhimurium survives in the tract compared to lower pH, which wasn¶t
Salmonella Typhimurium, where the higher and augmented the acid is, and the more influence it
can induce, while lower standards of pH values (at high pH value) somehow inhibit growth of
the bacterium. From reading this article, we concluded how strong, corrosive substances can
greatly affect a bacterium¶s growth rate and how the surrounding temperatures are also a key
factor for variables to be taken into consideration. It also occurred that we had questions whether
our target of E. coli can also be affected similarly in terms of the Salmonella Typhimurium. We
came to a thought of subjecting E. coli to a strong acid such as sulfuric acid as well as other acids
less in strength to compare the growth rates and potential alternations or dissimilarities. Another
article that we used was called "Comparative acid stress response of Listeria monocytogenes,
conditions." (Koutsoumanis, K. P., and J. N. Sofos. 321-326). This article greatly relates to our
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project because it includes reactions of various bacteria like Listeria monocytogenes, Escherichia
coli O157:H7 and Salmonella enterica serotype Typhimurium. All these bacteria are shown to be
conditions. This article elucidated how acidic environments provided a resistance against lethal
acidic conditions. Low pH habituation showed a different case where each response was
different of each pathogenic bacteria. Within the pH ranges of 5·0±6·0, 4·0±5·5 and 4·0±5·0,
Salmonella showed the most resistance among the rest. This article was ideal in giving a ³bigger
picture´ to our project. By showing the different kinds of working and failed bacterial
resistances, helped brought a slight conclusion to whether low pH values of chemical substances
can affect bacteria greatly. The pH variability shown in the article was also beneficial to our
project; for we are trying to find if low pH valued substances can influence Escherichia coli upon
growth response, rate or reactions, including values where the substances can inhibit growth if
possible or applicable. Our last article we took into consideration was "Adaptive responses of
Salmonella enterica serovar Typhimurium DT104 and other S. Typhimurium strains and
Escherichia coli O157 to low pH environments."(Jonge, R., W.S. Ritmeester, and F.M. Leusden.
625) So far we have discussed how the last three articles mentioned and explained the
comparative acidic conditions of several bacteria, the growth rates and response of Salmonella
Typhimurium, and three, how Acidic substances affect growth of anaerobic Salmonella. This
final article completes our variables we are searching for and is explain the adaptive responses of
the Salmonella bacterium. The article shows how Salmonella Typhimurium was subjected to
extreme acidic conditions. Using its current condition, it was compared to its growth and/or
adaptive response to that of acid-resistant E. coli O157 and other S. Typhimurium phage types.
At a certain phase of S. Typhimurium on the culture dish, the bacterium showed resistance at the
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pH of 2.5 hence concluding that the ability of S. Typhimurium to survive at pH value of 2.5 was
shown to be dependent upon other variables, considerably amino acids. This article was crucial
to our project because it provided the last of the three variables needed to be taken into
consideration; Salmonella Typhimurium adaptive response to acidic conditions itself. All four of
the professionally reviewed journal articles provided an essential outline for our project. In
provided the many variables for our project to be taken into consideration. As a result, our group
concurred to base our project upon Escherichia coli and its effects when subjected to low pH
à :
The focal purpose of this experiment, primarily, is to observe growth responses of Escherichia
coli affected by ph variability (mainly upon low pH valued substances such as sulfuric acid).
Upon this, we are to see the effects of the bacterium itself and address the problem statement or
question as: If low pH, caused by the addition of sulfuric acid with amalgamation of Tryptic Soy
Agar, cause changes in the growth rates of E. coli or if possible induce growth inhibition?
:
If Escherichia coli are subjected within sulfuric conditions at 37 degrees Celsius, then the
bacteria will most likely have its cell membrane ruptured in a way that it will cease growth or die
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:
mc Petri dishes (or more if needed for more test variables) ± provided by laboratory.
scientist/laboratory
mc Alternate nutrient Agar/Tryptic dishes (10 Petri dishes with agar) ± website (Science
Stuff)
à
:
Control group:
Test group:
Measurements:
- Bacterial growth will be algebraically analyzed and concluded through approximate formulas
Frequency:
Independent Variable:
- pH conditions applied.
Dependent Variable:
1.c Put on lab coat, latex gloves, and take out cotton swabs and Petri dishes.
2.c Fill up Petri dishes with necessary nutrients or use one that is already present.
3.c Using a sterilized cotton swap, place a small sample of E. coli (2 grams for adaption to
6.c Set ³C´ cultures with pH value of 7 using distilled water mixed with the nutrients.
.c Determine amount of sulfuric acid to be added to make the pH level of 0.5 using the
9.c Add the determined amount of sulfuric acid (10ml) to Petri dishes labeled ³pH ± 0.5´ and
10.cAt the same time, add the ³C´ control group Petri dishes in incubator at 37 degrees
Celsius.
12.cLeave the incubator for the time being and record any observations of initial growth rates.
13.cMonitor the growth from all Petri dishes for the duration of one month. Make sure to take
pictures of the cultures and to note any changes on the area on the Petri dish that the
culture has covered. In order to prevent any corrupted data, the cultures will be monitored
weekly.
14.cWhen one month has passed, carefully take out dishes and dispose upon supervision.
15.cAt the end of the experiment, use disinfectants, such as 70% ethanol, a 10% bleach
17.cCreate logarithmic graph or exponential delineating the average growth rate of each
bacterial culture.
1.cOnce calculated for each culture, plot the functions on a y ± axis versus x ± axis graph.
19.cCompare average graphs of ³C´ cultures to ³pH ± 0.5´ cultures and record observations.
20.cFinally, gather a final conclusion about E. coli growth rates and record in a research
report or table.
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? :
/releases/2010/02/10020210392.htm>.
Park, S.Y., et al. "The Combination of Zinc Compounds and Acidic pH Limits Aerobic Growth
of a Salmonella Typhimurium Poultry Marker Strain in Rich and Minimal Media." X
Dunkley, K.D., et al. "Growth and genetic responses of Salmonella Typhimurium to pH-shifts in
Jonge, R., W.S. Ritmeester, and F.M. Leusden. "Adaptive responses of Salmonella enterica
serovar Typhimurium DT104 and otherS. Typhimurium strains andEscherichia coli O157 to low
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