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Research Detail

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Sabiha Jahan Liya
Department of Mathematics and Natural Sciences, BRAC University, 66 Mohakhali, Dhaka-1212, Bangladesh

Romana Siddique*
Department of Mathematics and Natural Sciences, BRAC University, 66 Mohakhali, Dhaka-1212, Bangladesh

Urinary Tract Infection (UTI) is a worldwide phenomenon in modern times, in which the dependency on antibiotics for its treatment is increasing. The current study was conducted in order to find alternatives to antibiotics by investigating some commercial fruits for their antimicrobial activity. The fruits in this study included green apple (Malus domestica), papaya (Carica papaya), lemon (Citrus limon), and strawberry (Fragaria ananassa), which were used to prepare methanolic and ethanolic extracts through Soxhlet extraction technique. The extracts were used against bacteria that cause UTI, and five different strains were selected: E. coli (ATCC: 15922), E. coli (ATCC: 25922), Pseudomonas aeruginosa (ATCC: 27853), Enterococcus faecalis (ATCC: 29212), and Klebsiella pneumoniae. Antimicrobial tests of the extracts were conducted by following the agar well diffusion method, where ciprofloxacin was used as a positive control, and autoclaved distilled water was used as a negative control. Among the fruits, apple and papaya extracts did not show any zone of inhibition against any of the tested bacteria. However, both lemon and strawberry extracts showed inhibition zone against all of the mentioned bacteria. The ethanolic extracts of lemon and strawberry were more potent than their methanolic extracts. Lemon ethanolic extract showed the highest zone of inhibition against Pseudomonas aeruginosa (ATCC: 27853) (18.34 ± 0.58) and lowest one against Klebsiella pneumoniae (16.00 ± 1.00). Strawberry ethanolic extracts showed the highest zone of inhibition against Pseudomonas aeruginosa (ATCC: 27853) (16.33 ± 0.58) and the lowest one against Klebsiella pneumoniae (13.33 ± 0.58). As antibiotic resistance is paving the way for multidrug resistant bacteria, the results of lemon and strawberry can be considered to be used as an antimicrobial agent in treating urinary tract infections.

  UTI, Soxhlet, Methanolic extracts, Ethanolic extracts, Agar well diffusion
  Department of Mathematics and Natural Sciences, BRAC University, 66 Mohakhali, Dhaka-1212, Bangladesh
  
  
  Pest Management
  Fruit

We therefore aim to find out if the commercial fruits available in our country, like green apple, papaya, lemon, and strawberry, have the potency to exhibit antimicrobial activity against bacteria causing urinary tract infection.

Bacterial Strains. The following bacterial strains have been used for the current study: E. coli (ATCC: 15922), E. coli (ATCC: 25922), Klebsiella pneumoniae, Pseudomonas aeruginosa (ATCC: 27853), and Enterococcus faecalis (ATCC: 29212), which were collected from International Centre for Diarrhoeal Disease Research, Bangladesh (icddr,b). All the bacterial strains were maintained by weekly subculture. Chemicals and Reagents. The chemicals, agars, and antibiotic discs, which have been used in this experiment, belong to the following companies: Merck (New Jersey, United States), HiMedia (Mumbai, India) and Oxoid Limited (Cheshire, England), respectively. Preparation of Fruit Extracts. The fruits were bought fresh from the market and cut into thin slices. Then, they were dried under direct sunlight for 5–6 days and became crispy. Afterwards, they were ground into fine powder by using a grinder. 75 g of the ground powder was measured and subjected to Soxhlet extraction, where the ground powder is placed in the thimble, and the thimble is put in the chamber of Soxhlet apparatus. The extracting solvent is heated to a boiling point, and the vapors condense in the condenser. The condensed extractant then drips into the thimble containing the powder and extract it by contact. Afterwards, when the liquid reaches a certain level in the siphon tube, the solution containing the extract accumulates in a flask. 250 ml of absolute methanol and absolute ethanol were used as solvents to prepare methanolic and ethanolic crude extracts. Then, the solution was run through a rotary evaporator to evaporate the respective solvents, and the crude extracts were stored in autoclaved Macartney bottles. The crude extracts were later diluted to a concentration of 0.2 mg/μl and 0.6 mg/μl using the same respective solvents before conducting agar well diffusion. Agar Well Diffusion. The 24 h subcultured plates of bacterial strains were placed in the laminar hood and were used to make bacterial suspensions. The turbidity of the suspension was compared with the 0.5% MacFarland standard solution. Then, an autoclaved cotton swab was dipped into the suspension and rubbed horizontally across the surface of the labelled Mueller–Hinton agar (MHA) plates to conduct lawn culture of the bacterial strains. Then, a cork borer was used onto the MHA plates to create 3 wells on 3 different quadrants of the agar. After that, each well was labelled and filled accordingly with 60 mL of respective diluted methanolic and ethanolic fruit extracts and distilled water. Ciprofloxacin antibiotic disk was used as a positive control and placed onto one quadrant. Then, the MHA plates were kept in the incubator for 24 h at 37 °C, and the results were recorded the next day. All the tests were conducted 3 times to obtain the mean value of zones of inhibition, which was afterwards used to calculate the standard deviation value for the respective fruit extracts and the activity index for measuring the relative efficacy. Ethics. This study was performed in accordance to the good laboratory practice (GLP). No ethical approval was needed from the Institutional Review Board because there was no animal involved in conducting the study. ethanol extracts of apple gave zone of inhibition against Klebsiella sp. (9.2 mm) and Pseudomonas sp. (16.3 mm). According another study, both apple methanol and ethanol extracts showed inhibitory activity against S. aureus (14.3 mm and 13 mm, respectively) and EAEC (16.3 mm and 14.7 mm, respectively). But the extracts did not show any results against E. faecalis, P. aeruginosa or Klebsiella spp.. The unusual results in the current study may be due to the differences in the variety of apple used in this experiment compared to other studies. Antimicrobial properties are related to the bioactive compounds of the fruits, and apples are an important source of bioactive compounds like flavonoids, phenolic compounds, and anti-oxidants. Their concentrations and activity may vary with cultivar and variety. Papaya. In the current study, papaya extracts of methanol and ethanol did not show any zone of inhibition against any of the selected bacteria. According to one study, methanol extracts of Carica papaya leaf showed zone of inhibition against P. aeruginosa (17 mm), E. coli (12 mm), and Klebsiella pneumoniae (11 mm). However, the present study was conducted using only the ripened pulp of the papaya fruit, unlike other studies where its leaf or seed or peels were used. In another study, methanolic and ethanolic extracts of ripened papaya fruit pulp were used. The ethanol extract did not give any zone of inhibition for E. coli and P. aeruginosa. However, the methanol extract showed inhibition zone only against P. aeruginosa (11 mm) but not for E. coli. This variance in result may be due to the variety of the papaya used here. Also, the bacterial strains used in the experiments could be more resistant to the inhibitory effects of papaya fruit pulp extracts. Lemon. The methanolic and ethanolic extracts of lemon showed positive results for all of the bacterial strains tested. The results were shown as mean value ± the value of standard deviation. Lemon methanolic extract showed the highest antimicrobial activity against P. aeruginosa (ATCC: 27853) (16.67 ± 1.53) and the lowest antimicrobial activity against K. pneumoniae (14.67 ± 0.58). Lemon ethanolic extract showed the highest antimicrobial activity against P. aeruginosa (ATCC: 27853) (18.34 ± 0.58) and the lowest for K. pneumoniae (16.00 ± 1.00).

  European Journal of Microbiology and Immunology 8(2018)3, pp. 95–99
  
Funding Source:
1.   Budget:  
  

The use of antibiotics for treating any kind of infection or disease is very common these days. Their use often goes unmonitored or uncontrolled. Such a situation is giving rise to different multi-drug resistant bacteria as well, even in case of UTI. In this scenario, turning to nature is only inevitable, which, considering the advancement in the scientific research field, does not disappoint us. Lemon and strawberry showed antimicrobial potency against the tested bacteria according to this study; hence, looking for the alternatives using fruits is a dependable choice. All we have to do is to choose the right technique for identifying the right component to be used for treating the appropriate infection or disease.

  Journal
  


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