Antibacterial Activity of Paracetamol, Trimethoprim/sulfamethoxazole, Azithromycin, and Ciprofloxacin

Paracetamol have indirect antibiotic activity and are commonly coadministered with antimicrobial therapy. This study aimed to assess the synergistic effect of Paracetamol to with Azithromycin, Trimethoprim/sulfamethoxazole and Ciprofloxacin against salmonella and Escherichia coli. In this study, Ciprofloxacin, Trimethoprim/sulfamethoxazole and Azithromycin was mixed paracetamol against salmonella and Escherichia coli. The results showed that the interaction between Paracetamol and Azithromycin, Trimethoprim/sulfamethoxazole and ciprofloxacin exhibited more in inhibitory potency against on Gram-negative salmonella and Escherichia coli. The combinations of Paracetamol with the Azithromycin and Trimethoprim/sulfamethoxazole showed significant inhibitory activity on the salmonella and Escherichia coli comparing to the inhibition activity showed from the tested antibiotics alone.


Introduction
Acetaminophen or paracetamol chemically name N-acetyl-p-aminophenol is one of the most commonly used analgesic and anti-pyritic agents for alleviating acute and chronicpain.Due to its safety, acetaminophen is prescribed for patients in whom non-steroidal anti-inflammatory drugs (NSAIDs) are contraindicated, such as those with gastric ulcers and bronchial asthma, pregnant women, nursing mothers, and children (Leung, 2012;Roberts et al., 2016).It is typically used for mild to moderate pain.
Paracetamol is part of the class of drugs known as "aniline analgesics ". (Hanel and Lands 1982).It does not have significant anti-inflammatory activity and how it works is not entirely clear ( .Zainab R.Abdul-Hussein , 2014(.Paracetamol have indirect antibiotic activity and are commonly coadministered with antimicrobial therapy, it is important to understand the interactions between these two classes of drugs ( Zainab R Abdul-Hussein 2018 ) .This project had the following objectives with the hypothesis that the combinational therapy of an antibiotic and an Acetaminophen agent is viewed as an adjuvant molecule presents a viable therapeutic alternative to treat infections and inflammatory conditions.

Materials And Methods
This study, is based on evaluating the effectiveness of Acetaminophen or paracetamol in combination with some antibiotics against several strains of bacteria.

Tested Bacteria:
Escherichia coli and Salmonella typhimurium.

Preparation of bacterial sample:
Two isolates of Gram stain negative bacteria were included in this study, ( Escherichia coli and Salmonella typhimurium) both bacteria were obtained from private laboratory in Shahat city.

Antibacterial Assay:
Ciprofloxacin, Trimethoprim/sulfamethoxazole and Azithromycin put on nutrient agar medium plates, which their surfaces were previously streaked separately with tested E.Coli and also Ciprofloxacin, Trimethoprim/sulfamethoxazole and Azithromycin put on nutrient agar medium plates, which their surfaces were previously streaked separately with tested Salmonella typhimurium.The same procedures were done for tested paracetamol.
All plates were incubated at 37C° for 24 hours.After 24 hours the inhibition zones were measured in (mm) unit.

Assay of combinational effect of Paracetamol, Azithromycin, Ciprofloxacin and Trimethoprim/sulfamethoxazole
Each Ciprofloxacin, Trimethoprim/sulfamethoxazole and Azithromycin was mixed paracetamol put on nutrient agar medium plates, which their surfaces were previously streaked separately with tested E.Coli and also Ciprofloxacin, Trimethoprim/sulfamethoxazole and Azithromycin was mixed paracetamol put on nutrient agar medium plates, which their surfaces were previously streaked separately with tested Salmonella typhimurium.
All plates were incubated at 37C° for 24 hours.After 24 hours the inhibition zones were measured in (mm) unit.

Results and Discussion
In this study, results of antibacterial represent in The following Figures 1-4.When antibacterial Activity of Azithromycin+paracetamol, Ciprofloxacin alone, Ciprofloxacin +paracetamol and paracetamol alone on Escherichia coli, the results revealed that active performance against the tested bacteria; with mean of inhibition zones of 20 mm, 21 mm, 22 mm and 16mm, respectively.In addition Azithromycin no found any activity against Escherichia coli (figure 1).In other assay where a combination Trimethoprim/sulfamethoxazole +paracetamol had done and investigated for the antibacterial activity, the results showed inhibition zones of mean of 25mm against Escherichia coli (figure 2).

1-Antibacterial activity of paracetamol
In this study, paracetamol was tested separately for their antibacterial activity and showed antibacterial activity against salmonella Gram-negative, flagellated facultatively anaerobic bacilli bacterium and Escherichia coli Gram-negative, facultative anaerobic, rod-shaped, coliform bacterium.
Paracetamol, the most widely used are reported to improve the performance of antibiotics either through their synergistic antibacterial action with antibiotics or by reducing adherence, production of biofilm, and other virulence factors as well as by altering pathogen susceptibility to antibiotics.(Singh et al, 2021).
Paracetamol and/or non-steroidal anti-inflammatory drugs (NSAIDS) are of use for symptomatic relief.However, recent research suggests that some NSAIDs may also have the capacity to treat UTI caused by E. coli (Mazumdar K, Dutta NK. et al 2007).The effect of acetaminophen on the susceptibility of Pseudomonas aeruginosa against antibiotic was less significant than urinary tract bacteria.Up to date of this study, no study found done to determine the synergistic effects of paracetamol with Azithromycin, Trimethoprim/sulfamethoxazole, Ciprofloxacin against salmonella and Escherichia coli.This study is the first study investigate if there is any synergistic effect between the abovementioned drugs, and it revealed interesting evidence where it proved that the combination of Azithromycin , Trimethoprim/sulfamethoxazole, ciprofloxacin and Paracetamol have a good synergistic effect on salmonella and Escherichia coli , where it clarified that the antibacterial activity of paracetamol with Azithromycin, Trimethoprim/sulfamethoxazole combination is stronger than that given from Azithromycin and Trimethoprim/sulfamethoxazole, alone.These findings point to the possible adjuvant use of Azithromycin, Trimethoprim/sulfamethoxazole, ciprofloxacin and paracetamol in the treatment of salmonella and Escherichia coli infections.

Conclusion
This study conclude that paracetamol has a good synergistic effect with Azithromycin , Trimethoprim/sulfamethoxazole and ciprofloxacin against the Gram-negative salmonella and Escherichia coli, and it suggests that this combination could be considered as a good regimen to treat infection caused by these two species of pathogenic bacteria.In addition, this study recommends further studies to figure out the exact chemical reaction done between the tested antibiotic and paracetamol agents and stand behind this promising activity.

Fig. 2 :
Fig. 2: Antibacterial Activity of Trimethoprim/sulfamethoxazole alone and Trimethoprim/sulfamethoxazole +paracetamol on Escherichia coli In a next step the antibacterial activity of Trimethoprim/sulfamethoxazole alone, Trimethoprim/sulfamethoxazole+ paracetamol, Ciprofloxacin alone and Ciprofloxacin +paracetamol against both Salmonella was investigated and the result showed growth inhibition zones of mean of 25 mm, 27 mm, 23 mm and 23 mm respectively (figure 3).