The effect of seasonal variation on the antibacterial activity of Tithonia diversifolia (Hemsl.) A. Gray leaf extracts against pathogenic bacterial strains

doi:10.21203/rs.3.pex-2561/v1

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The effect of seasonal variation on the antibacterial activity of Tithonia diversifolia (Hemsl.) A. Gray leaf extracts against pathogenic bacterial strains

https://doi.org/10.21203/rs.3.pex-2561/v1

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The study investigates the antibacterial activity of Tithonia diversifolia leaves, including methanol, ethanol and water extracts, against Pseudomonas aeruginosa, Klebsiella pneumoniae, Serratia marcescens, Proteus vulgaris, Micrococcus luteus, Staphylococcus aureus, Bacillus subtilis, Clostridium sporogenes using the agar well diffusion technique. Minimum inhibitory concentrations (MIC) and minimum bactericidal concentrations (MBC) were performed on bacteria by a modification of the standard agar dilution method. Qualitative phytochemical analysis of leaf extract was determined by standard analytic methods. The study found that Klebsiella pneumoniae and Bacillus subtilis showed less susceptibility to water extracts, while Pseudomonas aeruginosa and Proteus vulgaris were sensitive to all water extract samples with zone of inhibition of 20.5 mm and 20.0 mm, respectively. However, all methanol extracts were susceptible to Pseudomonas aeruginosa, Proteus vulgaris, and Staphylococcus aureus with zones of inhibition of 18.0, 14.0, and 15.0 mm, respectively. Serratia marcescens showed less susceptibility to most methanol extracts, while Clostridium sporogenes showed susceptibility to all ethanol extracts. MICs and MBC values against bacterial strains ranged from 12 - 50 mgmL^-1 and 25 - 100 mgmL^-1 respectively. The study concluded that the leaves of Tithonia diversifolia contain phytochemicals with broad-spectrum antimicrobial effects, with ethanol extracts showing high sensitivity. The concentration of these phytochemicals varies by season, with higher levels observed during rainy seasons. The study highlights the potential of Tithonia diversifolia leaves for antimicrobial properties.

Antimicrobial activities

phytochemicals

Tithonia diversifolia

Minimum inhibitory concentrations (MIC)

Minimum bactericidal concentrations (MBC)

Antimicrobial resistance (AMR) has posed a significant threat to humanity since the discovery of antibiotics in the 19th century ^[1]. The Institute for Health Metrics and Evaluation ^[2] emphasizes the need for a comprehensive understanding of the resistance cost of Nigeria, particularly its emphasis on clinical medicine over environmental issues. Individual studies have produced knowledge on resistance in Nigeria. However, no coordinated efforts have been made to address this critical issue, as noted by the National Center for Disease Control and Prevention ^[3].

The efficacy of antibiotics against pathogenic bacteria is diminishing as a result of the emergence of antibiotic-resistant genes, which presents a substantial risk to both the environment and human well-being ^[4]. This requires ongoing efforts to discover and use effective antimicrobial substitutes to prevent the emergence or exacerbation of resistance ^[5].

Despite the numerous traditional plants and their derivatives used in disease care, only a few have been scientifically examined for their claimed effects. The potential therapeutic properties of the grey plant Tithonia diversifolia (Hemsl.) A, which is native to Africa and Central and South America, has attracted considerable interest ^[6]. Research has shown that its leaves possess antimicrobial properties, potentially treating certain infections ^{[7, 8, 9, 10]}. Additionally, their leaves possess phytochemical components, making them a prospective subject for future exploration in contemporary medicine ^{[11, 12, 13, 14]}.

The increasing literature on traditional medicine's use in treating bacterial infections and the increasing antimicrobial resistance among bacteria to certain antibiotics necessitates the survey and screening of antimicrobial compounds found in plants used in traditional clinical practices. Therefore, this study aimed to examine the effect of seasons on the antimicrobial properties of Tithonia diversifolia leaf extracts against specific bacterial isolates.

The leaves of T. diversifolia were air dried for three weeks at room temperature (20-22 ^,oC), ground into a coarse powder using an electric miller (Moulinex), and 50 grams of each pulverized sample were extracted using the Soxhlet extraction method with methanol, ethanol and water. The extracts were concentrated under reduced pressure using a rotary evaporator at 40 ^oC, reconstituted with 20% dimethyl sulfoxide (DMSO), and stored in the refrigerator at 4 ^oC until use.

Cultures were standardized using methods from the National Committee for Clinical Laboratory Standards [17]. A 0.2 mL of each 18-hour culture of each bacterium was suspended in universal sterile bottles containing 20 mL of sterile nutrient broth (Hi media) and incubated for 5 hours at 37 ° C to obtain a logarithmic growth phase. A sterile normal saline (0.9 %) was gradually added to compare turbidity with the McFarland standard of 0.5 which corresponds to approximately 1.0 × 10⁸ CFU/mL.

The antimicrobial activities of various extracts on test organisms were determined using agar well diffusion techniques. A 0.1 mL aliquot (optical density equivalent to 10⁷ -10⁸ CFU/mL) of logarithmic phase broth culture of each test organism was seeded in Mueller-Hinton agar medium (Hi-media) and dried in a dryer for 20 minutes. A standard cork borer (7 mm diameter) was used to cut uniform wells on the surface of the agar, and increasing concentrations of 0.15 mL of reconstituted test extract were added. Extracts were pre-incubated for 1 hour, and bacteria plates were incubated at 37 ° C for 18-24 hours. Zones of inhibition (mm) were measured and diluents were included as controls.

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The effect of seasonal variation on the antibacterial activity of Tithonia diversifolia (Hemsl.) A. Gray leaf extracts against pathogenic bacterial strains

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