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Introduction

Streptococcus faecalis is a bacteria that lives in the human digestive tract. At least 18 species of

this bacterium may be found within the human body. The majority of the microorganisms you

encounter will be of this kind. S. faecalis is a bacterium that is normally harmless when it is

located in the intestines and poses no threat to health as long as it stays there. Most cases of

septicemia, endocarditis, and meningitis are brought on by S. fecalis bacteria that have entered

the patient's bloodstream or urine. Gram-positive bacteria, including S. faecalis, have a thicker

cell wall and more discerning mechanisms for letting molecules and substances in and out of the

cell. The antibiotic resistance of S. faecalis is quite great. The organisms can generate ATP in the

presence of oxygen thanks to their facultative anaerobic nature, but it may rapidly convert to

fermentation if oxygen becomes scarce. The absence of catalase activity means hydrogen

peroxide cannot be broken down into water and oxygen. The answer becomes immediately clear

when hydrogen peroxide is introduced to the mixture. In conclusion, they earn their name,

streptococcus, since they are round and arranged in pairs or short chains.

Background

S. faecalis plays a pivotal role in glucose metabolism. Sugars and glycerol are examples of

carbohydrates. S. faecalis contains a phosphogluconate route and a glycolysis process known as

the Embden-Meyerhof-Parnas pathway. This bacteria can produce ATP without oxygen because

it can kick off fermentation using carbohydrates. Since the human digestive system is an oxygen-

deficient environment, this behavior is expected from the bacteria living there (Alghamdi &

Shakir, 2020). As a bonus, we can now relate our studies to the actual pathways present in these

bacteria. Since glucose is a carbohydrate that S. faecalis consumes during fermentation to digest

it and produce ATP for itself, this is most easily detected with the MR-VP Broth test. S. faecalis
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utilizes glucose and other carbs, which aid it when it has to undertake fermentation to gain

energy, allowing it to live much longer and better, even though it does not use any nitrate in any

manner, nor does it use acetoin. This might be because S. faecalis lives in a setting encouraging

evolutionary plasticity. This might be because S. faecalis lives in a setting encouraging

evolutionary plasticity.

Materials and Methods

You will need a Bunsen burner, a flame loop, a starch agar plate, an MRVP Broth tube, and a

Nitrate Broth tube to carry out this experiment. The process required that the organism be

inoculated using an aseptic technique and then streaked in a straight line over the medium on the

starch plate. The sample was then incubated for 24 hours, analyzed, subjected to an iodine test,

and examined again after 48 hours.

The four main steps of the gram staining procedure are as follows: To begin, a smear of the

bacteria culture that had been fixed with heat was stained with crystal violet, the primary stain.

The smear was then studied under a microscope. The germs were sterilized and then attached to

a microscope slide. Crystal violet is bound to iodide in the second stage, which causes the stain

to get entrapped on the cell wall. Third, quick decolonization was performed using a mixture of

acetone and ethanol. Counterstaining with safranin allowed the identification of the samples

(Medical Chemical Corporation, 2016). We used two test tubes labeled A and B to ensure our

observations were correct. The area, motility, and citrate tests were conducted using these test

tubes. Meanwhile, the nitrate and starch were each tested in their petri dishes. This table displays

the generated and documented findings. Microsoft Excel was utilized to do the statistical

analysis.
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Discussion

Because glucose was present in the MRVP broth, S. faecalis was able to ferment it and produce

the acid that was then used in the methyl red test to produce a positive result. Using this glucose

resulted in a broth with a pH of 4.4 or lower, which was necessary for a positive methyl red test

result. The VP test, which looks for acetoin, was also performed, but the result was negative

since the substance was red. This evidence suggests that S. faecalis were producing acetoin

(Alghamdi & Shakir, 2020). Acetoin is produced by S. faecalis when the bacteria are trying to

break down sugars like glucose without contributing significantly to the acidification of their

surrounding medium or broth. Because S. faecalis bacteria do not break down the starch in any

way or use it for energy synthesis, they failed the starch test and were, therefore, not allowed

inside the test cells. Because of this, S. faecalis did not pass the starch test. We know from

Ramsey's work that maltose was found in this cell, but it seems that S. faecalis does not need this

specific sugar.

Conclusion

The results of this study suggest that gram-positive bacteria predominate among the bacterial

species present. Compared to the nitrate, motility, starch, and citrate tests, the urea test resulted

in a much shorter turnaround time. Bacillus subtilis and other bacterial species became pink in

the Urea test and red in nitrate broth after first looking orange. The B. subtilis bacteria in the

petri dish took the shape of rods. The results allow me to conclude that Bacillus subtilis was

present in the sample and identified as the predominant bacterium.

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References

Alghamdi, F., & Shakir, M. (2020). The influence of Enterococcus faecalis as a dental root canal

pathogen on endodontic treatment: A systematic review. Cureus, 12(3).