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Journal of Plant Production and Sustainability DOI: https://doi.org/10.63072/jpps.25010 Volume 1, Issue 2, 2025, Pages 38-49 ISSN: 3135-7423 (Online) https://jplantps.org/ RESEARCH PAPER

Comparative in vitro evaluation of antimicrobial potential of ethanolic leaf extracts from different Artemisia species against infection-causing pathogens Hina Fazal1, Iram Fatima1, Gohar Rahman1, Abdul Haq1, Nisar Ahmad2*, Arshad Iqbal2 and Mohammad Ali2 1

Pakistan Council of Scientific and Industrial Research (PCSIR) Laboratories Complex, Peshawar 25120, Pakistan Center for Biotechnology and Microbiology, University of Swat, Swat-19200, Pakistan

*Correspondence author’s email: ahmadn@uswat.edu.pk Abstract Artemisia species are commonly administered in polyherbal drugs to treat various infectious diseases. The main component of Artemisia species is artemisinin that showed a multi-dimensional biological activity. Focusing on the therapeutic potential, the aim of this study was to provide scientific evidence for the antimicrobial activity of six Artemisia species viz. Artemisia scoparia, Artemisia persica, Artemisia arborescens, Artemisia absinthium, Artemisia vulgaris and Artemisia nilagirica. Leaf extracts were prepared using ethanol and then antimicrobial assay was performed following agar well diffusion method. Eight bacterial strains were grown on nutrient agar, however, C. albicans was grown in potato dextrose agar (PDA) followed by required incubation. Azithromycin, Ciprofloxacin and Clotrimazole were used as standard antibiotics for antimicrobial activity. Results revealed that all six Artemisia leaf extracts exhibited measurable antimicrobial activity, though with varying potency across

species. A. persica showed the strongest and broadest activity, producing the largest inhibition zones (up to 20.5 mm), particularly against E. coli, B. subtilis, K. pneumoniae, and S. mutans. A. scoparia and A. arborescens also demonstrated broad-spectrum inhibition, with moderate activity across all tested strains. A. absinthium and A. vulgaris displayed comparatively lower but consistent antimicrobial effects, with the greatest activity observed against P. aeruginosa and S. aureus, respectively. A. nilagirica exhibited selective activity, showing strong inhibition of S. aureus (17 mm) but no effect on P. aeruginosa, S. typhi, or C. albicans. Overall, the findings indicate that Artemisia species possess varying degrees of antimicrobial potential, with A. persica emerging as the most effective across the tested microorganisms. Hence, it can be concluded that the selected species can be used as a potential source of antimicrobial agents in food and pharmaceutical industries. © 2025 The Author(s) Keywords: Agar-well diffusion assay, Antimicrobial, Artemisia, Bacteria, K. pneumonia, Medicinal plants, S. mutans

Citation: Fazal, H., Fatima, I., Rahman, G., Haq, A., Ahmad, N., Iqbal, A., & Ali, M. (2025). Comparative in vitro evaluation of antimicrobial potential of ethanolic leaf extracts from different Artemisia species against infection-causing pathogens. Journal of Plant Production and Sustainability, 1(2), 38–49. https://doi.org/10.63072/jpps.25010

Introduction Plants have long been recognized as valuable sources of novel therapeutic compounds and have played a crucial role in traditional medicine systems (Parameswari et al., 2019; Khan et al., 2020). Bioactive plant-derived compounds exhibit significant antimicrobial and antitoxin properties, making them reliable candidates for use as antiinfective agents or as adjuncts to conventional therapies (Blaszyle & Holley, 1998). Nonetheless, it is essential to thoroughly investigate the pharmacological properties of these natural mixtures to understand their effects on biological systems (Viegas & Bolzani, 2006; Ushimaru et al., 2007). It is estimated that around 80% of the world’s medicines are derived from plant-based bioactive components (Owolabi et al., 2007). Due to their therapeutic potential, medicinal plants continue to attract considerable interest in modern pharmacological research

and drug discovery (Butler, 2004). Their use offers several advantages over synthetic drugs, particularly in addressing multi-drug resistance and minimizing adverse side effects, as herbal remedies are generally regarded as safer and less toxic alternatives (Keshebo et al., 2016; Nisar et al., 2017). Bacterial infections remain among the leading causes of mortality worldwide (Wu et al., 2022). Antibiotic therapy has long been a cornerstone of treatment; however, the misuse and overuse of antibiotics have accelerated the emergence of multidrug-resistant (MDR) bacterial strains (Prestinaci et al., 2015; Crits-Christoph et al., 2022). As a result, many conventional antibiotics and traditional treatments are becoming increasingly ineffective (Styers et al., 2006). Antimicrobial resistance (AMR) has therefore evolved into a serious global public health concern (Dupuis et al., 2022). It is projected that by 2050, infections caused by antibiotic-resistant pathogens could be responsible for approximately 10 million deaths annually, posing not only a health crisis but also a

https://jplantps.org/ significant economic burden (McEwen & Collignon, 2018; Antimicrobial Resistance Collaborators, 2022). The rise of MDR bacteria has severely restricted the clinical utility of existing antibiotics, as effective treatment options are diminishing or, in some cases, no longer available (Magiorakos et al., 2012). To address this challenge, the World Health Organization (WHO) released a global priority list in 2017 identifying antibiotic-resistant bacteria that pose the greatest threat to human health (De Oliveira et al., 2020). Despite the urgent need for new antibiotics, the discovery and development process remains hindered by high costs (often in the hundreds of millions of dollars), long development timelines (averaging around ten years), and limited commercial incentives only 17 novel antibiotics were approved between 2010 and 2021 (Safir et al., 2020; Berini et al., 2022). Consequently, there is growing scientific interest in exploring alternative therapeutic sources. Plant-derived natural products, with their rich diversity of bioactive compounds, have emerged as promising candidates for combating antibiotic-resistant pathogens (Subramani et al., 2017; Fatima et al., 2022). The Artemisia genus, which includes more than 500 species of herbs and shrubs distributed across the globe, represents the largest group within the family Asteraceae (Sengul et al., 2011; Ivănescu et al., 2021). Many species of this genus have long been valued in traditional medicine for their wide range of healing properties. They are rich in diverse secondary metabolites such as essential oils, flavonoids, organic acids, and lactones that exhibit significant pharmacological activities, including antimalarial, hepatoprotective, anticancer, and antiinflammatory effects (Bora & Sharma, 2011; Abad et al., 2012; Ekiert et al., 2021; Shinyuy et al., 2023). Among Artemisia species, Artemisia scoparia, Artemisia persica, Artemisia arborescens, Artemisia absinthium, Artemisia vulgaris, and Artemisia nilagirica, have a known history of geographical diversity and traditional use in different regions (Sengul et al., 2011; Russo et al., 2020; Bidgoli et al., 2021; Sharifi et al., 2022). These are commonly used to treat infections such as inflammation, cough cold, malaria, fever, influenza, and diabetes, digestive disorders and wound healing (Liu et al., 2009; Sharifi et al., 2022; Vitale et al., 2022). The demand for alternative antimicrobial agents, especially those derived from natural sources, has driven extensive research to evaluate plant-based drugs for combating infectious diseases (Vieira et al., 2017; Szczepanik et al., 2018; Guan et al., 2019). Antimicrobial potential of some Artemisia species have been reported in previous literature (Behbahani et al., 2017; Vieira et al., 2017; Szczepanik et al., 2018; Guan et al., 2019), however, no such detailed studies have been conducted on the comparison of antimicrobial activities of six selected Artemisia species. Hence, the current study encompasses both Gram-positive and Gram-negative bacteria, as well as fungal pathogens to assess the antimicrobial potential of the selected species.

Materials and Methods

Journal of Plant Production and Sustainability (2025) 1(2): 38-49 Plants collection The plants and seeds of different species were collected from different localities of Khyber Pakhtunkhwa Province, Pakistan. They were cultivated at Medicinal Botanic Garden, PCSIR, Peshawar. These species namely Artemisia scoparia, Artemisia persica, Artemisia arborescens, Artemisia absinthium and Artemisia vulgaris were successfully acclimatized at Garden and after reaching to stages of maturity they were collected and identified through taxonomic markers (Fig. 1). Their voucher specimens were prepared and deposited at Herbarium, PCSIR (PES), (Table 1) for future reference. Table 1 Voucher specimen numbers of Artemisia species at Herbarium, PCSIR, Peshawar (PES) S. No. Species Voucher 1 Artemisia scoparia 10696 2 Artemisia persica 10697 3 Artemisia arborescens 10698 4 Artemisia absinthium 10699 5 Artemisia vulgaris 10700 6 Artemisia nilagirica 10701 PCSIR = Pakistan Council of Scientific and Industrial Research

Extraction Fresh leaves of six species of Artemisia namely Artemisia scoparia, Artemisia persica, Artemisia arborescens, Artemisia absinthium, Artemisia vulgaris and Artemisia nilagirica were collected and washed with distilled water (Fig. 1). Leaves were dried in shade for 4-5 days and then powdered in blender. The powdered leaves of each species were weighed (Table 2) and extracted with ethanol using rotary evaporator. The extracts were dried under room temperature and the extract yield was measured using following equation: (

)

Subsequently, the dried extracts were dissolved in dimethyl sulfoxide (DMSO) at 100 mg mL-1 concentrations and filtered using Millipore syringe filter (0.2 µm pore size). The obtained extracts were stored at 4 °C for detailed analysis. Tested microorganisms The strains used in this study were American Type culture and collection (ATCC) strains viz., Escherichia coli (ATCC# 8738), Proteus mirabilis (ATCC# 12553), Streptococcus mutans (ATCC# 35668), Staphylococcus aureus (ATCC 6538) and clinical isolates including Klebsiella pneumonie, Salmonella typhi, Bacillus subtilis, P. aeruginosa and Candida albicans (yeast). The bacterial strains were grown in Nutrient Agar (Merck; Germany) and incubated at 37 °C for 24 h, however, C. albicans was grown in potato dextrose agar (PDA) and incubated at 35 °C for 24 h. Azithromycin, Ciprofloxacin, Clotrimazole were used as standard antibiotics for antimicrobial activity.

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Journal of Plant Production and Sustainability (2025) 1(2): 38-50

Table 2 Weight of leaf extracts and the extraction yield obtained from six selected Artemisia species S. No. 1 2 3 4 5 6

Species Artemisia scoparia Artemisia persica Artemisia arborescens Artemisia absinthium Artemisia vulgaris Artemisia nilagirica

Leaves weight (g) 10.48 ± 0.2a 10.19 ± 0.4a 3.76 ± 0.3c 10.10 ± 0.2a 10.10 ± 0.4a 5.90 ± 0.1b

Extract weight (g) 2.43 ± 0.04a 2.16 ± 0.03a 1.33 ± 0.05b 2.01 ± 0.01ab 1.69 ± 0.02b 1.27 ± 0.02b

Extract yield (%) 23.19 ± 2.1b 21.20 ± 1.8bc 35.37 ± 2.7a 19.90 ± 0.9c 16.73 ± 1.7d 21.52 ± 2.2b

Fig. 1 Pictorial presentation of different species of Artemisia (a) Artemisia scoparia (b) Artemisia persica (c) Artemisia arborescens (d) Artemisia absinthium (e) Artemisia vulgaris and (f) Artemisia nilagirica Antimicrobial assay Agar well diffusion method was used to evaluate antimicrobial activities of the leaf extracts (Haq et al., 2019). Inocula of the selected bacterial and fungal strains were prepared in normal saline (0.9%) and their turbidity was adjusted with McFarland standard (0.5). 100 μL volume of each microbial strains were swabbed on their

respective Muller Hinton agar (MHA) media plates. Wells were punched in MHA and sealed. About 100 μL of volume was taken from each leaf extracts and added in their respective wells. The culture plates were incubated at 37 °C for 24 h and then the antimicrobial activities were observed by measuring the diameter of zone of inhibition (mm). The tests were conducted in duplicate.

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Fig. 2 Antimicrobial of A. scoparia leaves extracts (100 µL) observed against nine microbial strains (a) E. coli (b) P. aeruginosa (c) K. pneumonia (d) S. typhi (e) P. mirabilis (f) S. mutans (g) S. aureus (h) B. subtilis (i) C. albicans.

Statistical analysis Three independent experimental replicates (n = 3) were performed for each species to obtain the mean values and standard deviations. Mean values that do not share a common letter are significantly different at p < 0.05. Letters in common indicate groups that are not significantly different based on the least significant difference (LSD) test.

Results

tested microbial strains, with inhibition zones ranging from 15.5 mm to 20.5 mm (Fig. 3; Table 3). The extract produced the largest zones of inhibition against E. coli and B. subtilis, each measuring 20.5 ± 2.2 mm and 20.5 ± 2.1 mm, respectively. Notable inhibitory effects were also observed for K. pneumoniae (19.5 ± 1.3 mm) and S. mutans (19.5 ± 1.5 mm), indicating strong responsiveness of these organisms to the extract. Moderate but consistent activity was recorded against P. aeruginosa (18.5 ± 1.7 mm), S. typhi (18.5 ± 1.3 mm), and S. aureus (18 ± 1.9 mm). Lower inhibition values were observed for P. mirabilis (15.5 ± 1.8 mm) and C. albicans (16.5 ± 0.6 mm), although both strains still exhibited measurable sensitivity to the leaf extract.

Antimicrobial potential of A. scoparia leaf extract The antimicrobial assessment of A. scoparia leaf extract revealed measurable inhibitory effects against all tested microbial strains, with inhibition zones ranging from 13.5 mm to 17.5 mm (Fig. 2). The extract produced inhibition zones of 15.5 ± 1.1 mm for E. coli, 17.5 ± 2.3 mm for P. aeruginosa, 16.5 ± 0.9 mm for K. pneumoniae, 15 ± 1.5 mm for S. typhi, 13.5 ± 2.1 mm for P. mirabilis, 17 ± 0.9 mm for S. mutans, 14.5 ± 1 mm for S. aureus, 16 ± 1.1 mm for B. subtilis, and 13.5 ± 0.7 mm for C. albicans, as presented in Table 3. These findings indicate that the leaf extract exhibits a broad antimicrobial spectrum, with varying degrees of growth inhibition observed across the bacterial and fungal species included in the study. Antimicrobial potential of A. persica leaf extract The antimicrobial evaluation of A. persica leaf extract demonstrated substantial inhibitory activity against all

Antimicrobial potential of A. arborescens leaf extract The antimicrobial assessment of A. arborescens leaf extract revealed a measurable inhibitory effect across all tested microorganisms, with zones of inhibition ranging from 11 mm to 17 mm (Fig. 4). The most prominent activity was observed against K. pneumoniae, which exhibited an inhibition zone of 17 ± 1.3 mm, representing the highest response among the evaluated strains. Moderate inhibition was recorded for B. subtilis (14 ± 1.2 mm), S. aureus (13.5 ± 1.3 mm), and P. aeruginosa (13 ± 1.0 mm), indicating consistent susceptibility to the extract. Additionally, inhibition zones of 12.5 ± 0.7 mm, 12.5 ± 1.1 mm, and 12.5 ± 1.1 mm were observed for E. coli, S. typhi, and S. mutans, respectively, reflecting comparatively lower but still notable antimicrobial effects. The least activity was detected against P. mirabilis (11.5 ± 1.1 mm) and C. albicans (11 ± 0.9 mm), which exhibited the smallest inhibition zones.

https://jplantps.org/ Antimicrobial potential of A. absinthium leaf extract The antimicrobial activity of A. absinthium leaf extract demonstrated a broad yet variable response across the tested microbial strains, with inhibition zones ranging from 10.5 mm to 15 mm (Fig. 5). The extract showed the strongest activity against P. aeruginosa, which produced an inhibition zone of 15 ± 0.9 mm, indicating a relatively high level of sensitivity to the phytochemicals present in the extract. Moderate inhibitory effects were recorded against K. pneumoniae (13 ± 0.9 mm), S. aureus (12.5 ± 0.4 mm), and B. subtilis (12.5 ± 0.3 mm), suggesting that these organisms exhibit consistent but slightly lower susceptibility. Similarly, inhibition zones of 11.5 ± 0.5 mm and 11.5 ± 0.4 mm were observed for E. coli and S. typhi, respectively, reflecting modest antimicrobial action. Reduced activity was noted against P. mirabilis (12 ± 0.3 mm), while the lowest inhibition zones, both measuring 10.5 mm, were recorded for S. mutans (10.5 ± 0.3 mm) and C. albicans (10.5 ± 0.4 mm), indicating limited responsiveness to the extract. Antimicrobial potential of A. vulgaris leaf extract The antimicrobial assessment of A. vulgaris leaf extract revealed a variable yet noteworthy inhibitory effect across the tested microbial strains. Overall, the extract produced inhibition zones ranging from 9.5 mm to 15.5 mm (Fig. 6). The strongest response was observed against S. aureus, which exhibited an inhibition zone of 15.5 ± 0.4 mm, indicating pronounced sensitivity to the bioactive constituents of the extract. This was followed closely by B. subtilis and P. mirabilis, both showing inhibition zones of 14.5 ± 0.3 mm and 14.5 ± 0.2 mm, respectively, suggesting that these organisms also respond strongly to the phytochemicals present. Moderate antimicrobial activity was recorded against C. albicans (13 ± 0.2 mm), while

Journal of Plant Production and Sustainability (2025) 1(2): 38-49 inhibition zones of 12.5 ± 0.3 mm and 12 ± 0.4 mm were noted for E. coli and P. aeruginosa, respectively, reflecting a modest but consistent level of sensitivity. Comparatively lower inhibition was observed in K. pneumoniae (11 ± 0.2 mm) and S. mutans (10.5 ± 0.3 mm), indicating reduced susceptibility. The least responsive strain was S. typhi, which produced the smallest inhibition zone of 9.5 ± 0.2 mm. Collectively the results summarized in Table 3 and depicted in Fig. 6 show that S. aureus, B. subtilis, and P. mirabilis are the most sensitive strains to A. vulgaris, while S. typhi demonstrates comparatively lower sensitivity. Antimicrobial potential of A. nilagirica leaf extract The antimicrobial evaluation of A. nilagirica leaf extract demonstrated a distinct pattern of activity across the tested microbial panel, with inhibition zones ranging from 0.00 mm to 17 mm. The strongest response was recorded against S. aureus, which exhibited an inhibition zone of 17 ± 0.9 mm, indicating notable susceptibility to the phytochemical components of the extract. Moderate inhibitory effects were observed against S. mutans and B. subtilis, each producing zones of 11.5 ± 0.2 mm, while K. pneumoniae showed slightly lower sensitivity with a zone of 12.5 ± 0.3 mm. A minimal inhibitory effect was detected in E. coli and P. mirabilis, both demonstrating inhibition zones of 9.5 ± 0.1 mm and 9.5 ± 0.3 mm, respectively. In contrast, A. nilagirica showed complete inactivity against P. aeruginosa, S. typhi, and C. albicans, as evidenced by the absence of any inhibition zones (0.00 mm), suggesting strong resistance or lack of responsiveness in these strains. These findings, summarized in Table 3 and illustrated in Fig. 7, highlight the selective antimicrobial capability of A. nilagirica, with particularly strong activity against S. aureus and limited or no activity against certain Gram-negative bacteria and fungal species.

Fig. 3 Antimicrobial of A. persica leaves extracts (100 µL) observed against nine microbial strains (a) E. coli (b) P. aeruginosa (c) K. pneumonia (d) S. typhi (e) P. mirabilis (f) S. mutans (g) S. aureus (h) B. subtilis (i) C. albicans

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Journal of Plant Production and Sustainability (2025) 1(2): 38-49

Fig. 4 Antimicrobial of A. arborescens leaves extracts (100 µL) observed against nine microbial strains (a) E. coli (b) P. aeruginosa (c) K. pneumonia (d) S. typhi (e) P. mirabilis (f) S. mutans (g) S. aureus (h) B. subtilis (i) C. albicans

Fig. 5 Antimicrobial of A. absinthium leaves extracts (100 µL) observed against nine microbial strains (a) E. coli (b) P. aeruginosa (c) K. pneumonia (d) S. typhi (e) P. mirabilis (f) S. mutans (g) S. aureus (h) B. subtilis (i) C. albicans Table 3 Antimicrobial activity observed in six selected Artemisia species against nine microbial strains Zone of inhibition (mm) Microbial A. A. A. A. A. A. persica Azith. Cipro. Clot. strains scoparia arborescens absinthium vulgaris nilagirica E. coli 15.5±1.1 20.5±2.2 12.5±0.7 11.5±0.5 12.5±0.3 9.5±0.1 20±1.1 35±2.6 P. aeruginosa 17.5±2.3 18.5±1.7 13±1.0 15±0.9 12±0.4 40±2.9 38±3.1 K. pneumoniae 16.5±0.9 19.5±1.3 17±1.3 13±0.9 11±0.2 12.5±0.3 23±1.8 S. typhi 15±1.5 18.5±1.3 12.5±1.1 11.5±0.4 9.5±0.2 32±1.9 42±3.2 P. mirabilis 13.5±2.1 15.5±1.8 11.5±1.1 12±0.3 14.5±0.2 9.5±0.3 41±2.7 42±2.9 S. mutans 17±0.9 19.5±1.5 12.5±1.1 10.5±0.3 10.5±0.3 11.5±0.2 31±2.2 50±3.2 S. aureus 14.5±1 18±1.9 13.5±1.3 12.5±0.4 15.5±0.4 17±0.9 38±2.5 35±2.5 B. subtilis 16±1.1 20.5±2.1 14±1.2 12.5±0.3 14.5±0.3 11.5±0.2 26±1.6 44±4.1 C. albicans 13.5±0.7 16.5±0.6 11±0.9 10.5±0.4 13±0.2 32.5±2.2 Escherichia coli (ATCC# 8738), Streptococcus mutans (ATCC# 35668), S. aureus (ATCC 6538), Proteus mirabilis (ATCC# 12553), Bacillus subtilis, Salmonella typhi, Klebsiella pneumonie, Candida albicans (Clinical isolates), Artemisia scoparia, Azithromycin (Azithro), Ciprofloxacin (Cipro), Clotrimazole (Clot), n = 3 for replicated independent experiment for each species.

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Journal of Plant Production and Sustainability (2025) 1(2): 38-49

Fig. 6 Antimicrobial of A. vulgaris leaves extracts (100 µL) observed against nine microbial strains (a) E. coli (b) P. aeruginosa (c) K. pneumonia (d) S. typhi (e) P. mirabilis (f) S. mutans (g) S. aureus (h) B. subtilis (i) C. albicans

Fig. 7 Antimicrobial of A. nilagirica leaves extracts (100 µL) observed against nine microbial strains (a) E. coli (b) P. aeruginosa (c) K. pneumonia (d) S. typhi (e) P. mirabilis (f) S. mutans (g) S. aureus (h) B. subtilis (i) C. albicans

Discussion In current study, antimicrobial potential of six Artemisia species were evaluated using some selected microbial strains. Among all species, A. persica showed highest activity as it produced highest inhibition zones against E. coli, B. subtilis, K. pneumonia and S. mutans. Our study correlates with the earlier findings of Bidgoli et al. (2021), who observed acceptable antifungal activity in A. persica essential oil against Aspergillus strains and thus, this specie can be used to prevent food crops from fungal

contaminations. Previously, researchers have reported antimicrobial potential in the essential oils of Artemisia species (Bouzenna et al., 2013; Aati et al., 2020). However, present study was designed to evaluate the antimicrobial activity of plant extracts for the first time. Said et al. (2016) collected aerial parts of Artemisia arborescens from different sites of the Mediterranean area (Southwestern Algeria and Southern Italy) and determined the antimicrobial activity. Similarly, Bouzenna and Krichen (Bouzenna et al., 2013) determined significant antifungal activity in the essential oil of A. arborescens against Rhizoctonia solani. The results indicated

https://jplantps.org/ that A. arborescens may be used as a valuable option in the control of the food-borne pathogens. Aati et al. (2020) examined antimicrobial activity of the isolated oils of A.absinthium and A. scoparia growing in Saudi Arabia using broth microdilution method, revealing that the essential oils isolated from the examined Artemisia species displayed growth inhibiting actions in a concentration-dependent manner on tested microorganism species. Our study also confirms the previous research findings by Cha et al. (2005); Sengul et al. (2011) who also reported significant antimicrobial potential in A. scoparia and A. absinthum against the examined strains. Hence, it can be inferred that the examined essential oils could be used to develop effective natural antimicrobial remedies with potential application in the fields of cosmetic industry, food manufacturing and medicine. In present study, A. nilagirica exhibited lowest antimicrobial potential and remain inactive against some of the tested strains which can be correlated with the earlier findings of Ahameethunisa and Hopper (2010) who recorded significant inhibitory potential in the methanol and hexane extracts of A. nilagirica against some grampositive and gram-negative bacteria. Furthermore, Hiremath et al. (2011) determined significant inhibitory activity of the aqueous and alcohol extract of Artemisia vulgaris indicating that the plant can fight these organisms effectively. Lou et al. (2011) investigated the antibacterial properties and mechanism of action of chlorogenic acid against various pathogenic bacteria. Their findings revealed that chlorogenic acid exhibited strong antibacterial activity, with minimum inhibitory concentration (MIC) values ranging from 20 to 80 μg/mL. The study further demonstrated that chlorogenic acid interacts with and partially disrupts the bacterial outer membrane, resulting in irreversible changes to membrane permeability and a slight leakage of nucleotides. Numerous studies have documented the antimicrobial potential of Artemisia species leaf extracts against a variety of human pathogenic bacteria and fungi (Cosoveanu et al., 2012; Juvatkar et al., 2012; Massiha et al., 2013; Parveen et al., 2014; Tajehmiri et al., 2014; Ghareeb, 2018; Alotibi & Rizwana, 2019; Parameswari et al., 2019; Benderradji et al., 2021). For instance, methanolic leaf extracts of A. campestris have demonstrated significant antibacterial activity against several human pathogens (Naili et al., 2010). Similarly, Al-Moghazy et al. (2017) reported that A. vulgaris leaf extracts possess antibacterial effects against multiple pathogenic bacterial strains. In contrast, studies examining the antimicrobial efficacy of Artemisia leaf extracts against phytopathogenic fungi and bacteria remain relatively limited (Carvalho et al., 2011; Mengane & Kamble, 2014; Nehad et al., 2017; Ma et al., 2019). Among the few available reports, Mengane and Kamble (2014) as well as Ma et al. (2019) observed that A. annua leaf extracts were highly effective against the phytopathogenic fungus Fusarium oxysporum. This strong antifungal activity may be linked to the chemical

Journal of Plant Production and Sustainability (2025) 1(2): 38-49 composition of A. annua (Potawale et al., 2008; Massiha et al., 2013). In particular, vitexin the second most abundant compound identified in A. annua extracts has been suggested to possess important antifungal properties (El-Khateeb, 2010).

Conclusion The results of this study highlight the significant antimicrobial potential of Artemisia species, indicating that their crude ethanolic extracts may serve as effective natural agents against various diseases caused by resistant microorganisms. Given that plant pathogens are a major constraint to crop productivity worldwide, these findings provide valuable guidelines for future research exploring other Artemisia species as potential sources of bioactive compounds effective against phytopathogens. The promising antimicrobial properties observed in this study suggest that Artemisia-derived extracts can be developed into eco-friendly alternatives to synthetic pesticides and antimicrobial drugs. Nevertheless, further investigations are needed to identify and characterize the specific active compounds responsible for these antimicrobial effects and to better understand their mechanisms of action. Future research should also focus on the endophytic microbial communities inhabiting Artemisia plants and the metabolites they produce, as these symbiotic microorganisms often generate potent bioactive substances that may enhance the antimicrobial efficacy of plant extracts. Combining Artemisia extracts with metabolites from associated endophytes could form the basis for innovative biological control strategies targeting specific phytopathogens. In addition, before practical applications can be recommended, comprehensive toxicological and clinical studies are necessary to evaluate the safety and efficacy of Artemisia-based formulations. Such efforts would support the development of standardized, effective, and safe herbal products that could serve as sustainable alternatives to conventional synthetic antimicrobial agents currently in use. Declarations i. Ethics approval and consent to participate Ethical approval and informed consent were not required for this study as it did not involve human participants, human data, or animals. ii. Consent for publication Consent for publication is not applicable. iii. Data availability All data generated or analyzed during this study are included in this article. iv. Competing interests Authors have declared that no competing interests exist.

https://jplantps.org/ v. Authors’ contributions H.F. executed experimental work; I.F. led the conceptualization of the study and guided the overall research design; G.R. managed the development of the methodology and supervised plant growth and stress-tolerance assays; A.H. prepared the initial version of the manuscript; N.A. handled statistical analyses and supported the interpretation of the findings; A.I. refined the manuscript through critical review and revision; M.A. assisted with data validation and ensured the accuracy and consistency of reported results. vi. Funding No funding was received for the design, data collection, analysis, interpretation, or writing of this research manuscript. vii. Acknowledgement Not applicable. viii. SDGs addressed Good Health and Well-Being (SDG 3); Industry, Innovation and Infrastructure (SDG 9); Responsible Consumption and Production (SDG 12)

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