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Method

A detergent is a specific surfactant or a specific mixture of surfactants associated with cleansing properties, when they are present in the dilute solutions. Detergents have been found to be associated with a long fatty acid chain, which allows the surrounding bacteria to get lysed. The compounds have been found to be specifically effective in the removal of dirt particles also, and sometimes these dirt particles are associated with bacteria also. The chelating process has been found to be effective in removing the unnecessary particles from a surface. Detergents have been found to be added into various disinfecting solutions associated with their lower surface tension and enhance the antibacterial effects. As stated in a research paper, it has been observed that the use of detergents in can reduce the growth of bacteria in dentin and also is responsible for killing of bacteria after 3 minutes of exposure (Al-Said and Ashgar 2020). The addition of detergents has been found to be associated with antibacterial activity against E. faecalis in dentinal tubules (AL-Salihi, 2019). A Zone of Inhibition Test, commonly known as a Kirby-Bauer Test, is a qualitative method for measuring antibiotic resistance in humans and for testing the capacity of solids and fabrics to prevent microbial growth in industry. A detergent is a specific surfactant or a specific combination of surfactants that has washing capabilities when present in dilute solutions. This test is used by researchers working on antimicrobial fabrics, surfaces, and liquids as a quick and easy technique to quantify and compare inhibitory activity levels. A sterile swab is used to disseminate roughly one million cells from a single strain onto an agar plate, which is subsequently incubated in the presence of the antimicrobial item (ex: an oxacillin disk). A zone of inhibition emerges on the agar plate if the bacterial or fungal strain is vulnerable to the antimicrobial agent, such as on the agar plate.

As per the information obtained from other research studies, it can be said that laundry detergent is responsible for killing of bacteria by lysing process. Since bacteria cell wall contains lipid and proteins, it can be easily lysed by the use of fatty acids, present in the detergents. In this way, the bacteria will die since cell wall lysis will occur. Sometimes, osmotic lysis also takes place due to the high concentration of detergents in the surrounding media of bacteria.

The method used for performing the experiment in the research study is Disk Diffusion Assay. This method has been used in order to measure the bacterial growth in presence and absence of detergents. The number of cells were counted for both the antibacterial (detergent) containing discs and normal discs. The detergent was diluted to the following dose values – 100%, 90%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 10%. These dilutions were used in order to analyse the growth of bacteria or killing of bacteria in the Petri plates. In other words, it can be said that the dilution of dosage values, specifically enables the researchers to observe the difference between the growth patterns in the above stated dilution values. This method of disc diffusion has been found to be used in other antibacterial analysis experiments also (Kourmouli et al. 2018). Thus, it can be said that the disk diffusion test is best for checking whether a specific antibacterial is imposing an inhibiting or killing effect on the growth of bacteria.

Results

The selected detergents of the research study will include – Dettol and Milton. The two groups of bacteria selected for the experimental study are – E. coli and S. aureus. These two bacteria were selected because they were found to specifically infect the human bodies and E. coli, already resides in the human guts.

Table 1: Dettol (detergent) and E. coli bacteria

Dettol E. coli

 Area of inhibition

 Area of inhibition

 Area of inhibition

 Mean

100%

28

30

29

29

90%

27

29

29

28.33333

80%

25

30

26

27

70%

25

28

27

26.66667

60%

24

26

24

24.66667

50%

23

25

24

24

40%

22

23

24

23

30%

20

22

22

21.33333

20%

19

20

20

19.66667

10%

19

19

20

19.33333

Table 2: Dettol and Staph bacteria

Dettol Staph A

 Area of inhibition

  Area of inhibition

  Area of inhibition

 Mean

100%

35

29

36

33.33333

90%

33

34

34

33.66667

80%

31

32

33

32

70%

29

30

29

29.33333

60%

27

26

27

26.66667

50%

24

25

26

25

40%

22

23

23

22.66667

30%

20

20

21

20.33333

20%

19

20

20

19.66667

10%

20

20

18

19.33333

Table 3: Milton and E. coli

Milton E. coli

 Area of inhibition

  Area of inhibition

  Area of inhibition

 Mean

100%

53.1

52.7

53.2

53

90%

42.4

45.7

49.4

45.83333

80%

41.9

42.6

47.3

43.93333

70%

41.6

42.1

45.9

43.2

60%

52.4

40.2

45.6

46.06667

50%

37.8

35.5

39.4

37.56667

40%

36.4

34.8

37.5

36.23333

30%

24.4

23.5

25.1

24.33333

20%

21.3

23.2

24.2

22.9

10%

20.6

19.7

0

13.43333

Table 4: Milton and S. aureus

Milton Staph A

 Area of inhibition

 Area of inhibition

 Area of inhibition

100%

57.4

58.2

55.8

57.13333

90%

55.7

53.7

52.9

54.1

80%

47.3

49.2

45.8

47.43333

70%

44.2

43.3

45.1

44.2

60%

43.8

42.7

42.8

43.1

50%

38.9

40.7

39.6

39.73333

40%

37.1

38.4

35.7

37.06667

30%

25.7

23.1

29.7

26.16667

20%

20.3

19.6

19.8

19.9

10%

19.6

0

20.5

13.36667

As observed from the above tables, it can be stated that the growth or E. coli, in Dettol is dose dependent. This means that the highest area of inhibition has been found to be associated with 100% Dettol and 10% Dettol has shown the lowest Area of inhibition in the Petri plates. There is a decreasing trend of the inhibition zones with the dilutions of the detergent. Thus, it can be stated that, lowest number of bacteria is present or highest number of bacteria is killed by the highest concentration of the detergent.

As observed from the above tables, it can be stated that the growth or S. aureus, in Dettol is dose dependent. This means that the zone of inhibition of bacteria has been found to be associated with 100% Dettol and 10% Dettol has shown the lowest Area of inhibition in the Petri plates. There is a decreasing trend of the inhibition zones with the dilutions of the detergent. Thus, it can be stated that, lowest number of bacteria is present or highest number of bacteria is killed by the highest concentration of the detergent.

As observed from the above tables, it can be stated that the growth or E. coli, in Milton is dose dependent. This means that the highest inhibition zones of bacteria have been found to be associated with 100% Milton and 10% Milton has shown the lowest Area of inhibition in the Petri plates. There is a decreasing trend of the bacterial inhibition zones with the dilutions of the detergent. Thus, it can be stated that, lowest number of bacteria is present or highest number of bacteria is killed by the highest concentration of the detergent.

As observed from the above tables, it can be stated that the growth or S. aureus, in Milton is dose dependent. This means tha the highest number of bacteria has been found to grow in 100% Milton and 10% Milton has shown the lowest Area of inhibition in the Petri plates. There is a decreasing trend of the area of inhibition with the dilutions of the detergent. Thus, it can be stated that, lowest number of bacteria is present or highest number of bacteria is killed by the highest concentration of the detergent.

concentration of the detergent

Fig 1: Graphical representation of the area of inhibition vs the dosage ranges of the detergents Dettol and Milton, chosen for the experiment. The decreased tendency of the graph has been observed for the increase in dilutions of the chemical detergents. This means that the area of inhibitions has decreased as the concentrations of chemical detergents went on decreasing. The box plot has shown that that the statistical data is not normally distributed. The data is not skewed since the data is distributed randomly around the error bars, in a decreasing order. The area of inhibition has been found to differ randomly, in a decreasing order. The same has also been represented by the tables where there is a difference in decreasing rate of the zones of inhibition as per the percentages Dettol and Milton concentrations.

When present in dilute solutions, a detergent is a specific surfactant or a specific blend of surfactants associated with washing capabilities. In other words, sodium salts of lengthy chain connected to alkyl hydrogen sulphate chains or even a long chain of benzene sulfonic acid can be found in a wide range of detergents. Surfactants and chelating agents are known to be present in detergents. This substance is used to remove dirt from dirty surfaces and to shift undesirable metal ions that may be found on a variety of surfaces and clothing.

Detergents have been discovered to be added to a variety of disinfecting solutions because they lessen surface tension and improve antibacterial properties. According to a research article, the use of detergents in may prevent the growth of germs in dentin and is also responsible for eliminating bacteria after 3 minutes of exposure (Niyonsaba 2019). The addition of detergents to dentinal tubules has been linked to antibacterial activity against E. faecalis (Pannerselvam 2021). According to information acquired from earlier research investigations, laundry detergent is responsible for the lysing process that kills germs.

As seen in the tables above, the development of microorganisms in Dettol and Milton is dosage dependant. This indicates that the maximum number of bacteria were discovered to grow in 100 percent diluted Milton, while the lowest Area of inhibition were found in 10 percent diluted Milton in Petri plates. The bacterial growth is diminishing when the detergent dilutions are increased. As a result, the lowest number of bacteria is present or the maximum number of bacteria is destroyed by the detergent at the highest concentration.

The decrease in growth of bacteria is thus associated with the killing of bacteria, at high concentrations of Dettol and Milton. On the other hand, it has been observed that Dettol has less inhibitory effect than Milton. In other words, it can be said that there is a very high reduction of inhibition zones between 100% Milton and 10% Milton. For Dettol, this reduction has been observed to be very less, however, a reduction in inhibition zones were observed. Zone of inhibition around paper discs, represents the fact that it has prevented the growth of bacteria in the zone or has killed every bacterium, which has grown there. In this way, circular zones of inhibitions were observed around the paper discs which are placed in the Petri plates. On the other hand, it has been observed that the paper discs associated with the Petri plates contain the chemical compounds, which have been found to create a major issue associated with the growth of bacteria (Sneha et al. 2021).

The results of the experiment can also be justified by evidence base. According to the findings of a research study, it has been found that bacteria are lysed by the presence of detergents and thus leads to the killing of the bacteria (Trisia, Philyria and Toemon 2018). On the other hand, it has also been observed that higher the strength of the detergent, higher is the number of killed bacteria, present inside the zones of inhibition (Ughamba, Erondu and Anyanwu 2019). The fact that the zone of inhibition is the representation of the fact that bacteria are getting killed in the Petri Plates. Another research study has stated that similar findings are observed with a drug’s effect on the growth of bacteria (Niyonzima 2019). In other words, it can be said that this experimental research study plays a major role in proving that detergents can be an agent, which can kill bacteria with proper dosage values. Moreover, it has been observed that the chemical compounds exerting lysing effect on bacteria, have been discussed specifically in many research studies (Ughamba, Erondu and Anyanwu 2019).

As per the findings from another research study, it can be stated that detergents are a major cost-effective source for killing bacteria (Yin et al., 2021). In other words, it can be said that the findings of the present experimental study can be stated to be justified. Since the current experiment has proved that there is a specific relationship between the growth of bacteria and the use of detergents as antibacterial agents, it can be stated that detergents will be effective in bringing out major changes in the killing process of a group of bacteria. Therefore, it can be said that the findings of the research study are specifically justified.

Conclusion

Detergents lower surface tension and boost antibacterial characteristics, they have been identified to be added to a range of disinfection solutions. According to a study, the use of detergents in dentin can help to inhibit the formation of germs and can also help to eliminate bacteria after 3 minutes of exposure (Sinta, P.H., Furtuna, D.K. and Fatmaria, F., 2020). Antibacterial activity against E. faecalis has been connected to the addition of detergents to dentinal tubules (Saeed, Iqbal and Ashraf 2020). Laundry detergent is thought to be important for the lysing process that kills bacteria, according to previous research findings. As can be seen from the above data section, E. coli growth in Dettol is dosage dependant. This indicates that 100 percent Dettol has the maximum area of inhibition, whereas 10 percent Dettol has the lowest area of inhibition on Petri plates. With increasing detergent dilutions, the inhibitory zones decrease in size. The same results were observed for the S.aureus and Milton detergent too. As a result, the lowest number of bacteria is present or the maximum number of bacteria is destroyed by the detergent at the highest concentration. In other words, it can be said that the experiment has addressed the aim and objectives in a very specific way. Moreover, it can be said that the overall conclusion of the experiment was also justified. However, it can be said that the deaths of bacteria were not confirmed. By the above statement, it is meant that it was only confirmed that bacteria present in the zones of inhibition were either dead or did not grow. Whether the detergent killed the growing bacteria, or it prevented their growth, cannot be determined by this experiment.

The experiment has a very specific drawback. There was no step performed to detect the dead bacteria present in the zones of inhibition. If dead bacteria are present in the zone of inhibition, then, it can be stated that bacteria were killed to form the zone. However, it was not detected whether the detergent was responsible for actually killing the bacteria or preventing its growth. This drawback can be solved in the future experiments by using a specific strategy to detect the dead cells, in order to prove whether the detergent dosages were associated with the killing of bacteria or not.

On a concluding note, it can be said that there is a specific growth preventing effect of different dosage ranges of the detergent on the selected groups of bacteria. In other words, it can be said that the experimental study has proved that the detergents can be used for the prevention of bacteria from growing on several surfaces.

References

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Al-Said, H. M., & Ashgar, S. S. (2020). A comparison of antibacterial efficacy of some household detergents available in Makkah, Saudi Arabia, against 16 ATCC bacterial strains and clinical isolates. African Journal of Microbiology Research, 14(8), 388-394.

AL-Salihi, S. S. (2019). Antibacterial Activity of Some Disinfectants and Detergents on some pathogenic bacteria. Journal of Pharmaceutical Sciences and Research, 11(2), 590-597.

Kourmouli, A., Valenti, M., van Rijn, E., Beaumont, H.J., Kalantzi, O.I., Schmidt-Ott, A. and Biskos, G., 2018. Can disc diffusion susceptibility tests assess the antimicrobial activity of engineered nanoparticles? Journal of Nanoparticle Research, 20(3), pp.1-6.

Kraft, L., Ribeiro, V.S.T., de Nazareno Wollmann, L.C.F., Suss, P.H. and Tuon, F.F., 2020. Determination of antibiotics and detergent residues in decellularized tissue-engineered heart valves using LC–MS/MS. Cell and Tissue Banking, 21(4), pp.573-584.

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Mian, A.H., Fatima, T., Qayyum, S., Ali, K., Shah, R. and Ali, M., 2020. A study of bacterial profile and antibiotic susceptibility pattern found in drinking water at district Mansehra, Pakistan. Applied Nanoscience, 10(12), pp.5435-5439.

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Niyonzima, F.N., 2019. Detergent?compatible bacterial cellulases. Journal of basic microbiology, 59(2), pp.134-147.

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