Microbiology lab

Adolat Beshimova

June 1, 2018

Saliva and Their Properties

Purpose

To find out if different sources of saliva will inhibit the growth of e.coli bacteria.

Hypothesis

dog saliva is known to have sterile properties and agents that directly attack the bacteria. The amylase
in the saliva of a dog can breakdown the cellular structure of a bacteria and ​The antibody IgA
(immunoglobulin A), which directly attacks bacteria in the saliva, ​due to these reasons we
hypothesize that the saliva of a dog is more effective than of human.

Materials

1. bunsen burner 8. 100 mg of ampicillin 15. electric balance

2. inoculating loops 9. marker 16.
3. Agar powder 10. laminar flow hood
4. 4 petri dishes 11. gloves
5. E. Colia bacteria 12. hole puncher
6. human saliva 13. micropipette tubes
7. dog saliva 14. weighing boat
Procedure

● obtain 4 sterile petri dishes

● Autoclave swabs
● Make agar solution: 0.230 grams of agar into a 100 ml of distilled water, boil until the solution
starts to bubbles
● Pour the hot agar solution into the the petri dishes in the laminar flow hood
● Make sure when pouring the agar solution into the petri dish, that the lid is half open not fully,
this will eliminate contamination
● Wait till the agar solution solidifies
● Make an antibiotic solution. 100 grams of ampicillin into 100 ml of water
● obtain human saliva as well as canis lupus familiaris saliva
● put 5 ml of solutions into 3 microcentrifuge tube
● label these solution
● Make paper discs out of filter paper
● put the 4 paper discs into each solution
● Using the bunsen burner, disinfect the inoculating loop
● Obtain e.coli
● Using the inoculating loop, spread the e.coli to the mother dish
● Wait till mother dish grows the bacteria
● Using the bacteria from the mother dish, spread the bacteria to other petri dishes too
● draw 2 vertical and horizontal lines evenly in the middle of of the petri dish, the line has to be
on the lid. These lines will create 4 sections where we will put the paper discs
● in each side of the corner, place one paper discs out of the one solution in to the petri dish
● label which side is which

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 ● repeat this step all over again for the other 3 petri dishes

Data

Table 1

trial 1 trail 2 trial 3 trial 4 average

ampicillin 10 mm 10.0 mm 9.0 mm 8.0 mm 9.25mm

dog saliva 3 mm 1 mm 2mm 3mm 2.25 mm

human saliva 0.5 mm 0mm 1 mm 1 mm 0.6mm

trial 1 trial 2 trial 3 trial 4 average

ampicillin 314 mm^2 314mm^2 254 mm^2 201 mm^2 271 mm^2

dog saliva 28mm^2 3.14 mm^2 13 mm^2 28 mm^2 18 mm^2

human saliva 0.8 mm^2 0mm^2 3.14 mm^2 3.14 mm^2

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Analysis

The data shows that there wasn’t any much effectiveness of dog saliva compared to
ampicillin, this might be due to human errors as well as other external factors. Some external factors
may include whether or not the dog ate just before the drool was obtained. Another reason why we
suspect it might have not worked is the mechanism of dog licking, when a dog licks a wound, it
removes all the bacteria of a wound. We did not stimulate this environment, on bacteria. We also
suspect the drool might have had some bacteria itself too. Experimental errors would be not washing
the incubator, not properly sterilizing the tools. Our control, ampicillin worked the best in terms of
zone of inhibition and human saliva did not significantly show inhibition according to data.
Ampicillin worked the best because it acts as an irreversible inhibitor of the enzyme transpeptidase,
which is needed by bacteria to make the cell wall. It inhibits the third and final stage of bacterial cell
wall synthesis in binary fission, which ultimately lead to the lysis of the cell that’s why ampicillin is
antibacterial.

Conclusion

Our research was inconclusive due to human errors. In terms of the result we obtained, it could be
noted that human salvia is not antibacterial in fact, according to our data, there were some instance
where the bacteria grew on the filter paper that contained the human saliva. If these steps were
repeated with more precautions and without human errors, there will be more accurate result.

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