Plants & Secondary Metabolites, Volume 11

Tannin Plants Saponin Microbes Health Dr. Madhuri Vajha ● Dr. Kugarthi Jayalakshmi ● Dr. Preeti Chaturvedi PLANTS & SECONDARY METABOLITES VOLUME XI Ambika Prasad Research Foundation, India Plants & Secondary Metabolites VOLUME 11 Dr. Madhuri Vajha Dr. Kugarthi Lakshmi Dr. Preeti Chaturvedi EDITORS Dr. Madhuri Vajha Department of Biological Sciences, G.B. Pant University of Agriculture & Technology, Pantnagar, Uttarakhand, India Dr. Kugarthi Jayalakshmi Department of biochemistry, Sri Padmavathi Mahila Viswavidyalayam, Tirupati, Andhra Pradesh, India Dr. Preeti Chaturvedi Department of Biological Sciences, G.B. Pant University of Agriculture & Technology, Pantnagar, Tanda Range, Uttarakhand, India Title: Plants & Secondary Metabolites, Volume 11 ISBN: 978-81-989192-9-8 Published by: Ambika Prasad Research Foundation An institutional publication Plants & Secondary Metabolites, Volume 11 First Edition: 2025 Copyright © Ambika Prasad Research Foundation The content of this book has been tried best to be provided with authenticated information. All the references necessary are listed. All attempts have been made to publish reliable information and acknowledge the copyright holders. If any copyright material(s) have not been acknowledged, please inform us so we may rectify it in our future reprints. Cover: Hexagonal structure Price: Rs. 1150/- Secondary metabolites: plant defense arsenal and humanity’s healing treasure. Preeti Chaturvedi Secondary metabolites are the plant’s hidden toolbox – tiny molecules that handle the big jobs of defense, signaling, and survival. Kugarthi Jayalakshmi Plants are nature’s master chemists, skillfully converting water, soil, and sunlight into a wealth of remarkable compounds—many beyond the imagination or capability of humankind to create. Secondary metabolites have regained prominence within pharmacognosy and phytochemistry due to their significant medicinal applications, including their roles as antibiotics, anticancer agents, and antioxidants, thus highlighting their renewed importance in natural product drug discovery. Madhuri Vajha Aditi Sunil Kale, Department of Botany, MG Vidyamandir’s L.V.H. Arts, Science and Commerce College Panchavati Nashik, Maharashtra, India Akshayjit Podder, Rupai M.E. School, Under BEEO, Hapjan, Department of School Education, Assam, India Alok Ranjan Sahu, Department of Botany, Vikash Degree College, Bargarh, Odisha, India Amardeep Kaur, S. D. College, Barnala, Punjab, India Bhagwati Prashad Sharma, Department of Botany, Sidharth Government College, Nadaun, Himachal Pradesh, India Brajesh Kumar Sahu, Department of Botany, P.M. College of Excellence, Government College, Vidisha, Madhya Pradesh, India Brijesh Kumar, Social Forestry Division, Bulandshahr, Uttar Pradesh, India Debangshu Agrahari, Department of Dravyaguna, Jeevak Ayurved Medical College & Hospital Research Centre, Chandauli, Uttar Pradesh, India Kadambini Das, University Department of Botany, Babasaheb Bhimrao Ambedkar Bihar University, Muzaffarpur, Bihar, India Kevileto Rote, Department of Soil & Water Conservation (Research, Demonstration and Training Centre) Kohima, Nagaland, India Manish Kumar, S. D. College, Barnala, Punjab, India Manvi Malwal, Post Graduate Government College for Girls 11 Chandigarh, India Paramita Ray, Ambika Prasad Research Foundation, Odisha, India Prasanta Kumar Samantray, P.G. Department of Botany, Shailabala Women’s Autonomous College, Cuttack, Odisha, India CONTRIBUTORS Rajkumari Supriya Devi, Ambika Prasad Research Foundation, Odisha, India Rajnikant Verma, Department of Botany, Govt. Girls College, Seoni Malwa, Narmadapuram, Madhya Pradesh, India Remya Krishnan, Daulat Ram College, Delhi University, Delhi, India Sanjeet Kumar, Ambika Prasad Research Foundation, Odisha, India Saraswati Majhi, Department of Life Sciences, Rama Devi Women’s University, Bhubaneswar, Odisha, India and P.G. Department of Botany, Shailabala Women’s Autonomous College, Cuttack, Odisha, India Satish S. Tambe, Department of Botany, MG Vidyamandir’s L.V.H. Arts, Science and Commerce College Panchavati Nashik, Maharashtra, India Smita Basole, Department of Botany, Balbhim College, Beed, Maharashtra, India Subhalakshmi Rout, Ambika Prasad Research Foundation, Odisha, India Subodh Kumar Kandari, Department of Botany, Omkarananda Sarashwati Government Degree College, Devprayag, Tehri Garhwal (Uttarakhand), India affiliated to: Sridev Suman Uttarakhand University, Badshahithaul, Tehri Garhwal, Uttarakhand, India Sugimani Marndi, Ambika Prasad Research Foundation, Odisha, India Sumitra Jethy, Ambika Prasad Research Foundation, Odisha, India Sweta Mishra, Ambika Prasad Research Foundation, Odisha, India Vikas Tailor, Department of Biology, Swami Vivekanand Government Model School, Block Asind, Bhilwara, Rajasthan, India PREFACE Plants have long stood at the intersection of nature and medicine, offering a silent yet powerful chemistry that continues to shape human survival and scientific curiosity. From ancient healing systems to modern pharmacological innovations, their secondary metabolites have emerged as key players in defense, communication and natural product research. This book, “Plants & Secondary Metabolites, Volume 11” is a compilation of research articles based on study of the phytochemicals and their pharmacology with relevance in a world facing mounting challenges of microbial resistance and ecological imbalance. This volume brings together eleven carefully curated chapters that explore a rich diversity of plant taxa from forest fruits and climbers to desert cacti and hemi-parasitic herbs, each contributing a unique profile of phytochemicals and bioactive potential. The studied species are wild and traditionally valued such as Smilax zeylanica, Carissa spinarum,Diospyros sylvatica,Gouania leptostachya, and Striga densiflora, to name a few. Included chapters revisit folk therapeutics and traditional knowledge systems, biochemical and antibacterial profiling while addressing the rising wave of antimicrobial resistance and reminding readers that plant based solutions may offer hope where conventional methods falter. This book serves as an invitation to observe more closely, to question more deeply, and to respect the intelligence embedded within plant systems. It is intended for researchers, students, conservationists, and scientists who seek interdisciplinary exploration between biodiversity and biotechnology, phytochemistry and pharmacology, curiosity and conservation. (Editors) BACKGROUND The accelerating loss of biodiversity, alarming rise of drug resistant microbes and lifestyle related diseases are compelling scientists to turn back to nature for answers. Plants produce special chemicals called secondary metabolites that help them survive in their environment. These same compounds have shown great potential in treating infections and other health problems. However, many useful plants and their chemical properties are still not fully studied or recorded. And, much of this biochemical diversity remains underexplored and insufficiently documented. In this context, “Plants & Secondary Metabolites” emerges as a timely contribution, bringing together multidisciplinary research focused on the phytochemical composition and bioactive potential of diverse plant species, including climbers, forest trees, wild fruits, desert flora, hemi-parasitic herbs and lesser known plant species. This editorial book is needed because many modern medicines are slowly losing their effectiveness due to increasing drug resistance, while new diseases and infections continue to emerge. At the same time, countless plant species with healing potential remain unexplored or poorly documented. Plants produce powerful secondary metabolites that can serve as natural sources of new drugs, yet information about these compounds is scattered and incomplete. By bringing together focused research on different plant species, their phytochemical composition, and their biological activity this particular volume would help fill an important knowledge gap. It aims to provide a single, reliable platform for students, researchers, and professionals to access updated information, support further studies, and encourage the sustainable use of plant based resources in healthcare and biotechnology. CONTENTS Chapter Title Authors Page 1 Evaluation of phytoconstituents and antibacterial activity of Smilax zeylanica L. (Smilacaceae) Saraswati Majhi, Prasanta Kumar Samantray and Sugimani Marndi 01 – 09 2 Study of microbial inhibition and secondary metabolites of Naringi crenulata (Roxb.) Nicolson (Rutaceae) Aditi Sunil Kale, Satish S. Tambe and Sugimani Marndi 10 – 17 3 Phytochemical screening and antimicrobial activity of Carissa spinarum L. (Apocynaceae): a wild therapeutic fruit Sugimani Marndi, Amardeep Kaur, Manish Kumar, Akshayjit Podder, Debangshu Agrahari, Brijesh Kumar, Manvi Malwal and Remya Krishnan 18 - 25 4 Desert bioactives: profiling phytochemicals and antibacterial potential in Cereus repandus (L.) Mill. (Cactaceae) Rajnikant Verma, Brajesh Kumar Sahu, Debangshu Agrahari, Smita Basole and Sugimani Marndi 26 – 32 5 Bioactive richness of the forest ebony: Diospyros sylvatica Roxb. (Ebenaceae) Alok Ranjan Sahu, Sugimani Marndi, Subhalakshmi Rout, Rajkumari Supriya Devi, Vikas Tailor and Sumitra Jethy 33 – 40 6 Biochemical wealth and microbial defense of the forest fruit Melia dubia Cav. (Meliaceae) Sugimani Marndi, Subhalakshmi Rout, Sumitra Jethy, Subodh Kumar Kandari, Kevileto Rote and 41 – 48 Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 6 Figure 2: Phytochemical analysis of S. zeylanica using n-hexane extract Figure 3: Phytochemical analysis of S. zeylanica using hydroethanolic extract Table 3: Estimation of Minimum Inhibitory Concentration (MIC) Hydroethanolic extract Concentration (mg/mL) MIC (mg/mL) 100 50 25 12.5 Smilax zeylanica(fruits) NG NG G G 50 Nutrient broth NG NG NG NG - Microbial broth G G G G - Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 7 (NG: No growth of bacteria; G: Growth) Figure 4: Minimum inhibition concentration (MIC) of fruits of S. zeylanica These observations indicated that the presence of diverse secondary metabolites in the hydroethanolic extract of S. zeylanica contribute to its antibacterial potential against E. coli. If the phytoconstituents can be studied further, they can be used in developing edible plant based therapeutic agents against gastrointestinal ailments like diarrhea, dysentery, and indigestion. CONCLUSION This study reinforced the ethnomedicinal significance of Smilax zeylanica, a plant long recognized in traditional medicine for its broad spectrum of healing properties. The detection of multiple bioactive compounds such as tannins, phenolics and alkaloids in the fruit extract confirmed its potential as a source of natural therapeutic agents. Although the antibacterial activity observed was moderate, it provides a scientific basis for the plant’s traditional applications in treating infectious and inflammatory conditions. Further quantitative and pharmacological studies are recommended for the isolation and characterization of the present active compounds responsible for these effects. Preserving and scientifically validating such traditional knowledge can contribute to the both sides: development of sustainable, Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 8 plant-based therapeutics and the conservation of indigenous medicinal heritage. REFERENCES Ahmad M, Khan MA, Zafar M and Sultana S. (2006). Treatment of common ailments by plant-based remedies among the people of District Attock (Punjab) of northern Pakistan. African Journal of Traditional, Complementary and Alternative Medicines. 4(1):112120. Dhanya SVS, Arbin A, Saema NGK, Sahana BK and Prashith KTR. (2018). Preliminary phytochemical analysis, antimicrobial and antioxidant activity of Smilax zeylanica L. (Smilacaceae). Journal of Drug Delivery and Therapeutics. 8(4): 237-243. García AC, Blancas J, Rodríguez BL, Binnqüist CL, Bahena HC, Calles AIM, Huelsz JAS and Medellín XL. (2021). How does urbanization affect perceptions and traditional knowledge of medicinal plants? Journal of Ethnobiology and Ethnomedicine. 17(1):48. Hossain AM, Saha S, Asadujjaman M and Kahan AS. (2013). Analgesic, antioxidant and antibacterial activity of Smilax zeylanica Linn. (family-Smilacaceae). Pharmacologyonline. 1:244-50. Jena N, Vimala, Singh B, Patra A, Sharma BP, Hossain E and Kumar S. (2025). Methods for ethnobotanical data collection, phytochemistry, antioxidant, anthelmintic, and antimicrobial activities for pharmacological evaluation of medicinal plants. Journal of Biodiversity and Conservation. 9(2): 87-107. Jena PK, Nayak BS, Dinda SC and Ellaiah P. (2011). Investigation on phytochemicals, anthelmintic and analgesic activities of Smilax zeylanica Linn. leafy extracts. Asian Journal of Chemistry. 23(10): 4307. Kamble A and Lobo V. (2022). Smilax zeylanica linn: pharmacognostic, taxonomical and phytochemical status. Herbal Science. 11(1): 1-14. Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 9 Kumar S, Mishra S, Mishra AK and Kumar SN. (2022). Floral Diversity of Koira & Barsuan Ranges of Bonai Forest Division, Odisha. Ambika Prasad Research Foundation, Odisha, India. Rizvi SAA, Einstein GP, Tulp OL, Sainvil F and Branly R. (2022). Introduction to traditional medicine and their role in prevention and treatment of emerging and re-emerging diseases. Biomolecules. 12(10): 1442. Sabarisenthil B and Kalaichelvan VK. (2017). A review on pharmacological activities of Smilax china and Smilax zeylanica. International Journal of Chemical and Pharmaceutical Sciences. 8(1): 57-64. Zhou Y, Zhou Z, Zheng L, Gong Z, Li Y, Jin Y, Huang Y and Chi M. (2023). Urinary Tract Infections Caused by Uropathogenic Escherichia coli: Mechanisms of Infection and Treatment Options. International Journal of Molecular Sciences. 24(13): 10537. Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 10 Chapter 2 Study of microbial inhibition and secondary metabolites of Naringi crenulata (Roxb.) Nicolson (Rutaceae) Aditi Sunil Kale1, Satish S. Tambe1 and Sugimani Marndi2* 1Department of Botany, MG Vidyamandir’s L.V.H. Arts, Science and Commerce College Panchavati Nashik, Maharashtra, India 2Ambika Prasad Research Foundation, Odisha, India *Email-Id:[email protected] DOI: https://doi.org/10.5281/zenodo.17421733 ABSTRACT In recent years, the global shift towards synthetic drug development has overshadowed the exploration of natural plant-based remedies, despite their long-standing contributions to traditional medicine. Naringi crenulata (Roxb.) Nicolson, a lesser-known member of the family Rutaceae, represents one such underexplored species with potential pharmacological importance. Present study aimed to evaluate the phytochemical composition and antibacterial activity of N. crenulata fruit extracts. Qualitative phytochemical screening revealed the presence of bioactive compounds such as tannins, flavonoids, alkaloids, phenols, and saponins predominantly in the hydroethanolic extract, while n-hexane extract showed limited metabolite presence. The antibacterial potential was assessed against Escherichia coli (MTCC 45) using the Minimum Inhibitory Concentration (MIC) method, demonstrating notable inhibition at lower concentrations in hydroethanolic extracts. These findings highlight N. crenulata as a promising source of natural bioactive compounds and support its potential role in developing plant based antibacterial agents. Keywords: Phytochemicals, antibacterial activity, traditional remedies INTRODUCTION Today, the mainstream science gives so much attention to synthetic drug discovery that the real spirit of studying natural products is fading away (Atanasov et al., 2021). The idea of discovering something new has veiled Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 11 the passion of researching the present resources we already have. While nature continues to rejuvenate the vast and largely unexplored library of flora amidst the growing pollution, human population, and destruction of natural habitats (Joppa et al., 2011) we, as a generation can take initiatives to study, understand and preserve the traditional knowledge with immense therapeutic potential that is dying a slow death. Naringi crenulata (Figure 1), the only recognized member of the family Rutaceae under genus Naringi, stands as one such promising species that deserves deeper scientific exploration. Figure 1: Fruits of Naringi crenulata Traditionally used in various indigenous healing systems, this plant is reputed for its medicinal properties against several ailments (Subramanianand Ramakrishnan, 2011). Its roots have been used as a remedy for Cobra bite (Sekhar et al., 2011), alleviating bodily pain (Pattanaiket al., 2008), gastric problems like colic, dysentery, and vomiting (Kumar, 2006; Ramachandran, 2010). Stem is useful in acne and anti-aging treatments (Mayuree, 2009). In folk medicine, bark is used for puerperal fever and pitta while leaves as a remedy for dysentery and epilepsy (Murty, 2010; Ramani, 2010). However, systematic studies validating these claims Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 12 remain limited. This experimental study was a step towards profiling the secondary metabolites present in its fruits and evaluating its antibacterial efficacy against pathogenic bacteria, which could bridge gap between traditional knowledge and modern science thereby, opening new pathways in natural product research. METHODOLOGY Fruits of Naringi crenulata were collected from the forests of ChandakaDampara Wildlife Sanctuary, Odisha, India. The plant species (Figure 2) was identified by authors followed by referring to the published literature (Kumar et al., 2022). Fruits were washed thoroughly under running tap water to remove soil particles and other surface contaminants. After cleaning, the fruits were air dried under shade at ambient room temperature to minimize thermal degradation of bioactive compounds. The dried material was coarsely powdered using a mechanical grinder and stored in airtight, sterile container to prevent moisture absorption and microbial contamination. Powdered fruit samples were subjected to solvent extraction using the maceration technique. Two solvent systems of differing polarity used for extraction were n-hexane (non-polar) and hydroethanol (7:3; polar) Figure 2: Fruits of Naringi crenulata Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 13 The extracts were filtered using Whatman No. 1 filter paper, and the filtrates were concentrated by air drying at room temperature until solvent residues were completely evaporated. The dried crude extracts were then stored under refrigerated conditions (4 °C) for two experimental setups following the standard procedures (Jena et al., 2025): a) Qualitative phytochemical screening to detect the presence of selected secondary metabolites, and b) Estimation of Minimum Inhibitory Concentration (MIC) to assess antibacterial activity against Escherichia coli (MTCC 45). RESULTS AND DISCUSSION The qualitative phytochemical screening of Naringi crenulata fruit extracts was done while keeping a clear view of solvent polarity on the secondary metabolite detection (Table 1; Figure 3-4). The non-polar n-hexane extract with polarity index of 0.1 appeared transparent and oily which showed the presence of only reducing sugars, indicating limited solubility of complex secondary metabolites in non-polar media. Table 1: Phytochemical screening of Naringi crenulata (fruits) Solvent Polarity Index Extract Color Detected Metabolites Extraction Efficiency Remarks n-Hexane 0.1 Transparent Reducing sugars Low High Extracted mainly lipophilic or simple carbohydrat es. Hydro ethanolic (7:3) 5.2 Pale green Tannins, Saponins, Flavonoids and Reducing sugars Extracted diverse bioactive compounds Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 14 The extraction efficiency was low; suggesting that n-hexane primarily extracted lipophilic or simple carbohydrate constituents. In contrast, the hydroethanolic extract (7:3), with a polarity index of 5.2, exhibited a palegreen coloration and tested positive for tannins, saponins, flavonoids, and reducing sugars. Figure 3: Phytochemical analysis of N. crenulata using n-hexane extract Figure 4: Phytochemical analysis of N. crenulata using hydroethanolic extract The hydroethanolic extract was comparatively richer in diverse bioactive metabolites, reflecting its superior capacity to dissolve both polar and moderately non-polar bioactive compounds. Then, the efficacy of the extract was screened for studying its antibacterial potential. The minimum inhibitory concentration (MIC) of the hydroethanolic extract of N. crenulata fruits was evaluated against Escherichia coli using serial dilutions ranging from 100 to 12.5 mg/mL (Table 2). Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 15 Table 2: Estimation of Minimum Inhibitory Concentration (MIC) Hydroethanolic Extract Concentration (mg/mL) MIC (mg/mL) 100 50 25 12.5 Naringi crenulata (fruits) NG NG NG NG 12.5 Nutrient broth NG NG NG NG - Microbial broth G G G G - Antibiotics NG NG NG NG - (NG: No growth of bacteria; G: Growth) It exhibited a dose dependent inhibitory effect, with no visible bacterial growth (NG) observed at any of the tested concentrations (100, 50, 25, and 12.5) mg/mL, indicating effective antibacterial activity even at the lowest tested concentration. The MIC value was determined to be 12.5 mg/mL (Figure 5), showing the minimum concentration required to inhibit visible bacterial growth. Figure 5: Minimum inhibition concentration (MIC) of fruits of N. crenulata These results suggested that N. crenulata fruit extract possesses significant antibacterial potential, particularly against E. coli and the potential of the fruits to serve as an effective natural remedy against common ailments Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 22 Solvent extraction: The powdered fruit material was subjected to solvent extraction using the Soxhlet method (Jena et al. 2025). Two solvent systems of differing polarity were employed to ensure broad-spectrum extraction of phytoconstituents: a) n-Hexane b) Hydroethanol (7:3) For each extraction, powdered material was placed in a thimble and was subjected to Soxhlet extraction using the two above mentioned solvents. The extracted material was collected and was placed in room temperature for evaporation of extra solvent. The remaining extracts were stored in sterile glass vials under refrigerated conditions (4 °C) until further experimental use. Qualitative phytochemical screening: Preliminary phytochemical screening of both n-hexane and hydroethanolic extracts was performed following standard protocols (Jena et al., 2025) to detect the presence of major classes of secondary metabolites. Determination of antibacterial activity: The antibacterial potential of the extracts was evaluated against Escherichia coli (MTCC 45) obtained from the Microbial Type Culture Collection, Chandigarh, India. The minimum inhibitory concentration (MIC) was determined using the broth microdilution method as described by Jena et al., (2025) with minor modifications. RESULTS AND DISCUSSION Depending on solvent polarity, the qualitative phytochemical screening of Carissa spinarum fruits was analyzed which detected the difference in secondary metabolite profiles (Table 2). Table 2: Detection of secondary metabolites of C. spinarum fruit extract Polarity Solvent extract Detected secondary metabolites Phytochemical richness Non-polar n-Hexane Reducing sugars Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 23 Polar Hydroethanolic Tannins, Saponins, Phenolic compounds and Reducing sugars ( = low, = high) The non-polar n-hexane extract showed a limited phytochemical spectrum where, reducing sugars was the only detected constituent, indicating a lower phytochemical richness. In contrast, the hydroethanolic (7:3) extract demonstrated a broader range of metabolites, showing tannins, saponins, phenolic compounds, and reducing sugars. The higher metabolite diversity in the polar extract suggested that polar solvents are more effective in extracting bioactive constituents from C. spinarum fruits. The antibacterial potential of the hydroethanolic extract of C. spinarum fruits was then evaluated against Escherichia coli by determining the minimum inhibitory concentration (MIC) (Table 3). Table 3: Estimation of Minimum Inhibitory Concentration (MIC) Hydroethanolic Extract Concentration (mg/mL) MIC (mg/mL) 100 50 25 12.5 Carissa spinarum (fruits) NG NG NG G 25 Nutrient broth NG NG NG NG - Microbial broth G G G G - Antibiotics NG NG NG NG - (NG: No growth of bacteria; G: Growth) The results showed that bacterial growth was completely inhibited (NG) at concentrations of 25 mg/mL and above, establishing MIC at 25 mg/mL (Figure 3). At lower concentrations (12.5 mg/mL), visible bacterial growth (G) was observed, confirming concentration dependent inhibitory activity. Control assays showed no bacterial growth (nutrient broth and antibiotic standards) validating the experimental conditions, whereas the microbial Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 24 broth control showed normal growth, confirming the viability of the bacterial inoculum. Figure 3: Minimum inhibition concentration (MIC) of fruits of C. spinarum CONCLUSION Rapid urbanization continues to exert pressure on wild habitats, while the rich knowledge of wild medicinal resources remains largely unexplored. In the present study, we profiled the phytoconstituents of Carissa spinarum fruits and assessed their phytochemical richness, which was further validated by determining the minimum inhibitory concentration (MIC) against Escherichia coli. Nevertheless, comprehensive scientific documentation, conservation of medicinal plants, and rigorous clinical evaluation are crucial to integrate traditional wisdom with modern medical science and to ensure the sustainable and responsible utilization of these bioresources in contemporary healthcare. REFERENCES Ansari IM and Patil DT. (2018). A brief review on phytochemical and pharmacological profile of Carissa spinarum L. Asian Journal of Pharmaceutical and Clinical Research. 11(9):12-18. Feyissa D and Melaku Y. (2016). Phytochemical, antibacterial and antioxidant studies of the leaves of Carissa spinarum. International Journal of Chemistry and Pharmaceutical Sciences.7: 25-30. Jena N, Vimala, Singh B, Patra A, Sharma BP, Hossain E and Kumar S. (2025). Methods for ethnobotanical data collection, phytochemistry, Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 25 antioxidant, anthelmintic, and antimicrobial activities for pharmacological evaluation of medicinal plants. Journal of Biodiversity and Conservation. 9(2): 87-107. Kumar S. (2022). Medicinal plants. Intech Open, United Kingdom. Nishteswar K. (2014). Depleting medicinal plant resources: a threat for survival of ayurveda. Ayu. 35(4): 349-350. Sharma N, Kumar V, Gupta N, Shekhar P and Kaur PS. (2023). Traditional importance, phytochemistry, pharmacology, and toxicological attributes of the promising medicinal herb Carissa spinarum L. Separations.10(3):158. https://doi.org/10.3390/separati ons10030158 Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 26 Chapter 4 Desert bioactives: profiling phytochemicals and antibacterial potential in Cereus repandus (L.) Mill. (Cactaceae) Rajnikant Verma1, Brajesh Kumar Sahu2, Debangshu Agrahari3, Smita Basole4 and Sugimani Marndi5* 1Department of Botany, Govt. Girls College, Seoni Malwa, Narmadapuram, Madhya Pradesh, India 2Department of Botany, P.M. College of Excellence, Government College, Vidisha, Madhya Pradesh, India 3Department of Dravyaguna Vijnana, Jeevak Ayurved Medical College and Hospital Research Centre, Chandauli, Uttar Pradesh, India 4Department of Botany,Balbhim College, Beed, Maharashtra, India 5Ambika Prasad Research Foundation, Odisha, India *Email-Id:sugimani2marn[email protected]m DOI: https://doi.org/10.5281/zenodo.17452212 ABSTRACT Cereus repandus, a succulent tree, has long been recognized in traditional medicine for its potential health benefits. Present study aimed to profile the phytochemical composition and evaluate the antibacterial activity of its hydroethanolic fruit extract. Qualitative phytochemical analysis revealed the presence of primary metabolites, including proteins, carbohydrates, and amino acids, highlighting the nutritional value of the fruit. Secondary metabolites such as tannins, flavonoids, and reducing sugars were detected, indicating potential pharmacological properties. The antibacterial activity was assessed against Escherichia coli, which established a minimum inhibitory concentration (MIC) of 50 mg/mL. The extract exhibited a concentration dependent inhibition of bacterial growth, suggesting moderate antibacterial potential. The observed bioactivity is likely associated with the presence of secondary metabolites, particularly tannins and flavonoids. These findings support the therapeutic property of C. repandus and highlight its potential as a source of bioactive compounds for future pharmacological applications. Keywords: Antibacterial, ethnomedicine, nutraceuticals Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 27 INTRODUCTION Cacti are remarkable desert plants that have adapted to survive under extreme environmental conditions, storing water and nutrients within their succulent tissues (Hultineet al., 2023). Many cactus species are valued for their nutritional and medicinal properties. Edible parts such as fruits and cladodes (flattened stems) are rich in nutrients including carbohydrates, amino acids, vitamins, and minerals. They also contain a wide range of bioactive compounds such as flavonoids, tannins, alkaloids, and phenolics that contribute to its pharmacological properties (Conte et al.,2025). Traditionally, cacti have been consumed as food and used in folk medicine across arid regions of the America, Africa, and Asia for treating ailments like diabetes, obesity, and digestive disorders (Monteiroet al.,2023). Native to the coasts of Venezuela to Colombia in the Caribbean Sea (POWO, 2025), Cereus repandus (Figure 1), commonly called as the Peruvian apple cactus or Cadushi by the locals, thrives in typical dry tropical habitats. Figure 1: Fruit of Cereus repandus attached to the plant Its interchangeable taxonomic identity often gets confused with the name C. peruvianus L. In southern part of Brazil, its cladodes formulation is known to inhibit gastric lesions and its extract is also marketed as a herbal Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 28 medicine used for weight loss (Nunes et al.,2022). Its stems are reviewed (Morton, 1967) to be used as a staple food anddried stems to be used as torches, fuel and fences and when matured, thick trunks are also used for carpentary. In recent years, scientific researches have also highlighted cacti as functional foods with potential health values. Their ethnobotanical use has also been linked to today’s nutraceutical and pharmaceutical applications. In that light, fruits of C. repandus (Figure 2) were screened for studying their phytochemical richness and antimicrobial activity against Escherichia coli, a gram negative bacteria associated with common gastrointestinal disorders (Petersen, 2022). METHODOLOGY The fruits of Cereus repandus were collected from nearby Athagarh areas of Cuttack district, Odisha, India. The plant species was identified by the authors (Figure 1). Figure 2: Collected fruit sample of C. repandus The Soxhlet extraction method was adopted using hydroethanolic extract for qualitative phytochemical analysis (Table 1). Detection of primary and Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 29 secondary metabolites was conducted and antibacterial activity was studied by determining the minimum inhibitory concentration (MIC) using Escherichia coli using standard methods. (Devi et al.,2023; Jena et al.,2025; Sharma et al.,2025). RESULTS AND DISCUSSION The hydroethanolic extract of Cereus repandus fruits was analyzed for the presence of primary and secondary metabolites (Table 1). The extract contained primary metabolites including proteins, carbohydrates, and amino acids (Figure 3), indicating the nutritional and metabolic potential of the fruit. Secondary metabolites such as tannins, flavonoids, and reducing sugars were also detected (Figure 4), confirming the presence of key phytochemicals that may contribute to the pharmacological efficacy of the fruit. Table 1: Qualitative phytochemical analysis of Cereus repandus for detection of primary and secondary metabolites Extract Category of Metabolites Detected Bioactive Compounds Significance Hydroethanolic Primary Metabolites Proteins, Carbohydrates and Amino acids Indicates the nutritional and metabolic potential of the fruit. Secondary Metabolites Tannins, Flavonoids and Reducing sugars Confirms the presence of key phytochemicals contributing to pharmacological efficacy. Figure 3: Detection of primary metabolites Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 30 Figure 4: Detection of secondary metabolites These findings suggest that C. repandus possesses both nutritional and bioactive properties, supporting its traditional and potential therapeutic applications. The antibacterial potential of the hydroethanolic extract was evaluated against Escherichia coli using the broth microdilution method (Table 2). The extract inhibited bacterial growth at concentrations of 50 mg/mL and above, establishing the minimum inhibitory concentration (MIC) at 50 mg/mL (Figure 5). At lower concentrations (25 and 12.5 mg/mL), visible bacterial growth was observed, indicating a concentration dependent effect. Control experiments validated the results, with nutrient broth showing no growth and microbial broth demonstrating normal bacterial proliferation, while antibiotic controls exhibited complete inhibition. These results suggest that the hydroethanolic extract of C. repandus fruits possesses moderate antibacterial activity, likely attributable to the detected secondary metabolites, such as tannins and flavonoids, known for their antimicrobial properties. Table 2: Estimation of Minimum Inhibitory Concentration (MIC) Hydroethanolic Extract Concentration (mg/mL) MIC (mg/mL) 100 50 25 12.5 Cereus repandus (fruits) NG NG G G 50 Nutrient broth NG NG NG NG - Microbial broth G G G G - Antibiotics NG NG NG NG - (NG: No growth of bacteria; G: Growth) Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 31 Figure 5: Determination of minimum inhibitory concentration (MIC) CONCLUSION The hydroethanolic extract of Cereus repandus fruits bespeaks dual significance: nutritional and bioactive. The presence of primary metabolites shows its metabolic and dietary value, while secondary metabolites, notably tannins and flavonoids, contribute to its moderate antibacterial activity against E. coli. This study validates the traditional use of C. repandus and highlights its potential as a natural source of bioactive compounds with pharmacological relevance. Further studies exploring other microbial strains and in vivo applications could expand its therapeutic potential and support its integration into modern medicinal frameworks. REFERENCES Conte G, MinhósYI, Moraes EM, Costa FBD and Franco FF. (2025). Secondary metabolites of Cactaceae: current knowledge and perspectives. Discover Plants. 2: 243. Devi RS, Satapathy KB and Kumar S. (2023). Validation of tribal claims for formulation of future drugs through evaluation of ethnopharmacological values of Ludwigia adscendens. Medicinal Plants. 15(4): 691-697. Hultine KR, Hernández HT, Williams DG, Albeke SE, Tran N, Puente R and Larios E. (2023). Global change impacts on cacti (Cactaceae): Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 38 Figure 3: Detection of secondary metabolites in n-hexane extract Figure 4: Detection of secondary metabolites in hydroethanolic extract Table 2: Estimation of Minimum Inhibitory Concentration (MIC) Extract Type Tested Organism Range of Concentrations (mg/mL) MIC (mg/mL) Antibacterial Strength Hydroethanolic extract (Diospyros sylvatica fruits) E. coli 12.5 – 100 100 Moderate Antibiotic control E. coli Standard - Strong Nutrient broth - - - Negative Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 39 Figure 5: Determination of minimum inhibitory concentration (MIC) CONCLUSION The results of this study revealed that Diospyros sylvatica is not only a symbol of ecological endurance but also a reservoir of valuable phytochemicals. The presence of diverse bioactive compounds and noticeable antibacterial activity indicates that this species holds great promise for developing natural remedies and plant based antibacterial agents. By bridging traditional knowledge with modern science, the study reaffirms the medicinal potential of forest ebony and underlines the need for its conservation and further pharmacological research. REFERENCES Devi RS, Satapathy KB and Kumar S. (2023). Validation of tribal claims for formulation of future drugs through evaluation of ethnopharmacological values of Ludwigia adscendens. Medicinal Plants. 15(4): 691-697. Fankam AG, Nchiozem-Ngnitedema VA, Seukep AJ, Bitchagno GTM and Kuete V. (2025). Ethnopharmacology, phytochemistry and pharmacological potential of the genus Diospyros. Journal of Ethnopharmacology. 120763. Jena N, Vimala, Singh B, Patra A, Sharma BP, Hossain E and Kumar S. (2025). Methods for ethnobotanical data collection, phytochemistry, antioxidant, anthelmintic, and antimicrobial activities for Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 40 pharmacological evaluation of medicinal plants. Journal of Biodiversity and Conservation. 9(2): 87-107. Plants of the world online (POWO). (2025). Diospyros sylvatica Roxb. Royal Botanic Gardens Kew, United Kingdom. Rao CG, Sri BS, Lakshmi BS, Babu YR and Padal SB. (2024). GC-MS analysis and screening of antimicrobial potentialities of the medicinal plants Diospyrossylvatica Roxb. And Diospyroschloroxylon Roxb. Innovations. 79: 1133-1152. Ribeiro A, Serrano R, da Silva IBM, Gomes ET, Pinto JF and Silva O. (2023). The Genus Diospyros: A review of novel insights into the biological activity and species of Mozambican flora. Plants. 12(15):2833. Vaou N, Stavropoulou E, Voidarou C, Tsigalou C and Bezirtzoglou E. (2021). Towards advances in medicinal plant antimicrobial Activity: a review study on challenges and future perspectives. Microorganisms.9(10):2041. Wallnöfer B. (2001). The biology and systematics of Ebenaceae: a review. Annalen des Naturhistorischen Museums in Wien. Serie B für Botanik und Zoologie. 1:485-512. Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 41 Chapter 6 Biochemical wealth and microbial defense of the forest fruit Melia dubia Cav. (Meliaceae) Sugimani Marndi1, Subhalakshmi Rout1, Sumitra Jethy1, Subodh Kumar Kandari2, Kevileto Rote3 and Sanjeet Kumar1* 1Ambika Prasad Research Foundation, Odisha, India 2Department of Botany, Omkarananda Sarashwati Government Degree College, Devprayag, Tehri Garhwal (Uttarakhand), India affiliated to: Sridev Suman Uttarakhand University, Badshahithaul, Tehri Garhwal, Uttarakhand, India 3Department of Soil & Water Conservation (Research, Demonstration and Training Centre) Kohima, Nagaland, India *Email-Id:s[email protected]. DOI: https://doi.org/10.5281/zenodo.17466821 ABSTRACT Plants have long served as valuable sources of natural therapeutic agents, especially in traditional medicinal systems. Melia dubia Cav. (Meliaceae), commonly known as the forest neem or Malabar neem was explored in this study to screen the phytochemical composition and antibacterial activity of its fruits by using solvent extraction method. Qualitative phytochemical screening revealed that the hydroethanolic extract contained several bioactive compounds, including tannins, saponins, flavonoids, terpenoids, reducing sugars and alkaloids, whereas the non-polar extract did not detect any tested metabolites. Antibacterial activity assessed against Escherichia coli demonstrated complete growth inhibition at 100 mg/mL concentration of the hydroethanolic extract, confirming moderate antibacterial potential. These findings support the ethnomedicinal relevance of M. dubia and highlight its potential as a source of natural antibacterial compounds for future pharmacological development. Keywords: Pharmacological value, phytochemicals, traditional medicine Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 42 INTRODUCTION Medicinal plants have been an integral part of human healthcare since ancient times, providing essential remedies for various diseases (Mahalakshmi et al., 2020; Haji et al., 2024; Susheela et al., 2008). These ethnobotanical knowledge from across the world forms the foundation for many modern drug discoveries, particularly those addressing bacterial infections and resistance. Many contemporary pharmaceuticals have originated from plant derived compounds that were first identified through traditional medicinal practices (Nasim et al., 2022). In recent years, the rise of antibiotic resistance has renewed global interest in ethnomedicine for exploring novel antimicrobial agents from natural sources (Anand et al., 2019). This exploration of plant based bioactive compounds gives a promising alternative to synthetic drugs which is sustainable and less expensive. Melia dubiais one such multipurpose tree belonging to family Meliaceae, native to tropical and subtropical regions of Asia and Africa (Ramanan et al., 2024). Traditionally, various parts of this plant including fruits, leaves, bark, and seeds have been used to treat ailments such as colic, skin infections, fever, asthma, malaria, and venereal diseases (Table 1). Table 1: Ethnomedicinal uses of different parts of Melia dubia Plant part Traditional medicinal uses Mode of use Geographical reference Fruit Used as an anthelmintic, astringent, and for treating colic, skin diseases, sores, and scabies (Mahalakshmi et al., 2020; Haji et al., 2024; Susheela et al., 2008). Green fruits are made into a paste for treating skin ailmentsand fruit juice mixed with sulphur used for scabies. India, Philippines (Laguna) Leaves Used to treat stomachache, asthma, eczema, and malaria; also as mosquito repellent and pesticide (Haji et al., 2024). Boiled leaves used orally for stomachache; burnt leaves used for repelling mosquitoes; mashed leaves mixed with kerosene applied on skin for acne, allergies, and ringworm. India, Philippines (Mindanao), Local tribal practices Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 43 Stem Bark Applied for skin ailments, open wounds (Haji et al., 2024). Decoction or extract of bark applied topically or consumed. Indonesia (West Timor), Philippines Seeds Used as insect repellent (Haji et al., 2024). Oil extracted for topical or fumigant use. Philippines Wood Valued for furniture, farm tools, and carpentry due to its durable quality (Goswami et al., 2020). Processed wood material. India and Southeast Asia Whole Plant Employed traditionally to treat leprosy, eczema, asthma, malaria, fevers, venereal diseases, diabetes cholelithiasis, acariasis, and for reducing pain (Mahalakshmi et al., 2020; Haji et al., 2024; Susheela et al., 2008). Decoctions, poultices, or powders prepared from entire plant parts. India, Africa, Southeast Asia Figure 1: Fruits of Melia dubia The plant is also valued for its durable timber and its ecological adaptability to dry deciduous habitats (Goswami et al., 2020). Despite its extensive Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 44 traditional and economical use, the phytochemical and antibacterial potential of M. dubia fruits remain underexplored. Therefore, this study was undertaken to qualitatively assess its phytochemical composition and evaluate the antibacterial activity of its hydroethanolic fruit extract against Escherichia coli, a common multifaceted pathogenic bacterium responsible for various gastrointestinal diseases (Braz et al., 2020). METHODOLOGY Mature fruits of Melia dubia were collected from the natural forest stands of Chandaka, Odisha, India and identified by the authors. The samples were given tags and were put in sterile cotton bags (Figure 2). Figure 2: Collected fruit sample of Melia dubia for experimental use Then, the fruits were washed thoroughly to remove debris and surface contaminants. They were extracted using the maceration method (Devi et al., 2025) with two solvents of different polarity: n-hexane and hydroethanol (1:1). The extracts were concentrated at room temperature to remove residual solvent and stored in sterile vials at 4 °C until further use. Preliminary phytochemical screening of both extracts was carried out to detect major classes of secondary metabolites. Then, the antibacterial Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 45 potential of the extracts was assessed against Escherichia coli (MTCC 45) and the minimum inhibitory concentration (MIC) was determined using the broth micro dilution method with minor modifications to the standard protocols (Jena et al., 2025). RESULTS AND DISCUSSION The qualitative phytochemical analysis of Melia dubia (Table 2) revealed distinct variations in the metabolite profiles depending on the polarity of the extraction solvent. Table 2: Qualitative phytochemical analysis of M. dubia Polarity Extract Detected secondary metabolites Non-Polar Polar n-Hexane No tested metabolites were detected Hydroethanolic Tannin, Saponin, Flavonoids, Terpenoids, Reducing sugars and Alkaloids The non-polar n-hexane extract showed negative result for all the tested secondary metabolites (Figure 3). In contrast, the polar hydroethanolic extract exhibited a rich phytochemical composition, confirming the presence of tannins, saponins, flavonoids, terpenoids, reducing sugars, and alkaloids (Figure 2). These compounds are known to possess diverse pharmacological properties (Leela et al., 2016), including antimicrobial, antioxidant, and anti-inflammatory activities. The antibacterial potential of the hydroethanolic fruit extract was hence, evaluated through the determination of its Minimum Inhibitory Concentration (MIC) against Escherichia coli (Table 3). The extract exhibited complete inhibition of bacterial growth at the concentration of 100 mg/mL, while lower concentrations (50, 25, and 12.5 mg/mL) allowed visible bacterial growth. The nutrient broth control showed no growth, confirming sterility, whereas the microbial broth exhibited consistent bacterial proliferation. The antibiotic control displayed complete inhibition, validating the assay performance. Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 46 Figure 3: Detection of secondary metabolites in n-hexane extract Figure 3: Detection of secondary metabolites in hydroethanolic extract Table 3: Estimation of Minimum Inhibitory Concentration (MIC) Sample Extract type Bacterial response Minimum Inhibitory Concentration (MIC) Melia dubia (fruit) Hydroethanolic Inhibited growth at 100 mg/mL; growth observed at lower concentrations (50, 25, 12.5) mg/mL 100 mg/mL Nutrient Broth - No growth (sterile medium) - Microbial Broth - Consistent bacterial growth - Antibiotic (standard) - Complete inhibition (positive control) - Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 47 These observations indicated that Melia dubia fruit possesses moderate antibacterial activity, with hydroethanolic extracts being effective than the non-polar extract due to their higher secondary metabolite content and this validates its traditional medicinal practices. CONCLUSION The findings of present study revealed that the fruits of Melia dubia are rich in diverse secondary metabolites such as tannins, flavonoids, terpenoids, alkaloids, and saponins those are recognized for their pharmacological significance. The hydroethanolic extract exhibited marked antibacterial activity against Escherichia coli, showing inhibition at the tested concentration of 100 mg/mL. These outcomes lend scientific support to the traditional medicinal uses of M. dubia and showcase its promise as a natural reservoir of bioactive molecules with antibacterial potential. To strengthen these observations, future research should focus on quantitative profiling, purification, and characterization of individual phytochemicals to better understand the pharmacological relevance of this valuable yet understudied species. REFERENCES Anand U, Jacobo-Herrera N, Altemimi A and Lakhssassi N. (2019). A Comprehensive review on medicinal plants as antimicrobial therapeutics: potential avenues of biocompatible drug discovery. Metabolites. 9(11): 258. Braz VS, Melchior K and Moreira CG. (2020). Escherichia coli as a multifaceted pathogenic and versatile bacterium. Frontiers in Cellular and Infection Microbiology. 10: 548492. Devi RS, Satapathy KB and Kumar S. (2023). Validation of tribal claims for formulation of future drugs through evaluation of ethnopharmacological values of Ludwigia adscendens. Medicinal Plants. 15(4): 691-697. Goswami M, Bhagta S and Sharma D. (2020). Melia dubia and its importance: a review. International Journal of Economic Plants. 7(Feb 1): 29-33. Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 54 Preparation of plant material: The freshly collected fruits (Figure 2) were thoroughly washed with running tap water to remove adhering soil and surface contaminants. After cleaning, the fruits were shade dried for a short period to eliminate surface moisture while preserving natural metabolites. The partially dried material was then coarsely chopped and macerated directly for extraction without prior pulverization to minimize the loss of volatile or heat-sensitive constituents (Jena et al., 2025). Extraction procedure: The extraction of bioactive compounds was carried out by cold maceration using solvents of differing polarity to ensure broad spectrum recovery of phytoconstituents using standard protocol (Devi et al., 2023). Approximately 50 g of fresh fruit material was soaked separately in n-hexane (non-polar) and hydroethanol (7:3, polar) in sterile conical flasks. The mixtures were kept for 72 hours at room temperature with occasional stirring to enhance solvent penetration. Figure 2: Collected fruits of Ceriscoides turgida for experimental use After the extraction period, the mixtures were filtered through Whatman No. 1 filter paper to separate the liquid extracts from plant residues. The filtrates were then air dried at ambient temperature to allow complete Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 55 solvent evaporation. The concentrated crude extracts were weighed and stored in airtight, sterile vials under refrigerated conditions (4 °C) until further use. Phytochemical screening: Both n-hexane and hydroethanolic extracts were subjected to qualitative phytochemical analysis using standard phytochemical assays to identify the presence of key secondary metabolites such as tannins, saponins, terpenoids, phenolics, flavonoids, alkaloids, and reducing sugars. The intensity of coloration or precipitate formation in each reaction was used as an indicator of compound presence (Jena et al., 2025). Assessment of antibacterial activity: The antibacterial efficacy of the extracts was assessed against Escherichia coli (MTCC 45), a standard laboratory strain obtained from the Microbial Type Culture Collection (MTCC), Chandigarh, India. The minimum inhibitory concentration (MIC) was determined using the broth microdilution technique with serial dilutions of the extracts prepared in nutrient broth following standard methods (Jena et al., 2025). Each assay was performed and bacterial growth was monitored visually after incubation at 37°C for 24 hours. The lowest concentration showing complete inhibition of visible growth was recorded as the MIC value, indicating the minimum inhibitory concentration of the extract against E. coli. RESULTS AND DISCUSSION A clear contrast was observed between the extracts obtained with non-polar and polar solvents (Table 3). The n-hexane extract showed negligible phytochemical content, indicating its ineffectiveness in isolating key constituents from the fruit material. In comparison, the hydroethanolic extract (1:1) displayed a broad spectrum of secondary metabolites, including tannins, saponins, flavonoids, terpenoids, phenolics, reducing sugars, and alkaloids. The substantial recovery of metabolites in the hydroethanolic medium reflected that the solvent has capacity to dissolve compounds of varying polarity, therefore, captured a wider range of bioactive molecules (Figure 3 and 4). Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 56 Table 3: Detection of secondary metabolites in fruit extract of C. turgida Solvent Extract Polarity Number of metabolites detected Major phytochemical Classes Extraction efficiency n-Hexane Nonpolar 0 None Poor Hydroethanol (1:1) Polar 7 Tannins, Saponins, Flavonoids, Terpenoids, Phenolics, Reducing sugars, Alkaloids Good Figure 3: Phytochemical analysis of non-polar extract Figure 4: Phytochemical analysis in polar extract Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 57 The antibacterial potential of the hydroethanolic extract was tested against Escherichia coli (MTCC 45) using the broth dilution method (Table 4). Noticeable inhibition of bacterial growth occurred only at the highest concentration (100 mg/mL), while lower concentrations (50, 25, and 12.5 mg/mL) showed no inhibitory effect (Figure 5). Table 4: Estimation of Minimum Inhibitory Concentration (MIC) Hydroethanolic Extract Concentration (mg/mL) MIC (mg/mL) 100 50 25 12.5 Ceriscoides turgida (fruits) NG G G G 100 Nutrient broth NG NG NG NG - Microbial broth G G G G - Antibiotics NG NG NG NG - (NG: No growth of bacteria; G: Growth) Figure 5: Minimum inhibition concentration (MIC) of fruits of C.turgida Control assays behaved as expected, confirming the validity of the results. The observed inhibition at higher doses suggests that the extract contains compounds with mild antibacterial potential, which may act synergistically Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 58 at sufficient concentration. Although the activity was modest, it supports the ethnomedicinal relevance of C. turgida. CONCLUSION Modern drug discovery often focuses on speed and uniform results, sometimes forgetting the long history of traditional healing practices. Ethnomedicine is not just a list of herbal cures; it carries the knowledge and experiences of many generations. By combining this traditional wisdom with modern scientific research, we can develop safer and more sustainable medicines while also protecting valuable plant diversity. The frequent and consistent use of Ceriscoides turgida among different tribal communities shows its trusted healing value. In this study, the hydroethanolic fruit extract of this plant showed strong phytochemical richness and a moderate antibacterial activity with minimum inhibitory concentration (MIC) at 100mg/mL, supporting its traditional use. The future of medicine may depend on learning from such traditional systems, turning old knowledge into new discoveries that connect nature with science. REFERENCES Bodele SK, Shahare NH and Dange SP. (2017). Physicochemical and Phytochemical Investigation on the Root of Ceriscoides turgida (Roxb.) Tirveng. Journal of Chemical and Pharmaceutical Research. 9(4): 69-73. Davies MK and Hollman A. (2002). Stamps in cardiology: Quinine. Heart. 88(2):118. Desborough MJR and Keeling DM. (2017). The aspirin story - from willow to wonder drug. British Journal of Haematology. 177(5): 674-683. Devi RS, Satapathy KB and Kumar S. (2023). Validation of tribal claims for formulation of future drugs through evaluation of ethnopharmacological values of Ludwigia adscendens. Medicinal Plants. 15(4): 691-697. Jain SK and Srivastava S. (2003). Some folk herbal medicines for possible use in veterinary practices. Indian Journal of Traditional Knowledge. 2(2): 118-25. Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 59 Jena N, Vimala, Singh B, Patra A, Sharma BP, Hossain E and Kumar S. (2025). Methods for ethnobotanical data collection, phytochemistry, antioxidant, anthelmintic, and antimicrobial activities for pharmacological evaluation of medicinal plants. Journal of Biodiversity and Conservation. 9(2): 87-107. Krishna S, Bustamante L, Haynes RK and Staines HM. (2008). Artemisinins: their growing importance in medicine. Trends in Pharmacological Sciences. 29(10): 520-527. Mahapatra AD, Bhowmik P, Banerjee A, Das A, Ojha D and Chattopadhyay D. (2019). Ethnomedicinal wisdom: an approach for antiviral drug development. In: New look to phytomedicine. Academic Press. Saxena HO and Brahmam M. (1995). The Flora of Orissa, Volume 2. Regional Research Laboratory, Bhubaneswar & Orissa Forest Development Corporation Limited, Bhubaneswar, Odisha, India. Shukla R and Harmukh N. (2020). Gardenia turgidaroxb. (Kharhar) an important medicinal plant of Chhattisgarh, India. International Journal of Advance Research and Innovative Ideas in Education. 6(4):2395-4396. Telles S, Pathak S, Singh N and Balkrishna A. (2014). Research on traditional medicine: what has been done, the difficulties, and possible solutions. Evidence Based Complementary and Alternative Medicine. 2014: 495635. Zhang Q. (2015). Traditional and Complementary Medicine in Primary Health Care. In: Health For All: The Journey of Universal Health Coverage. Orient Blackswan, Hyderabad, Telangana, India. Zilani MN, Sultana NA, Bakshi MK, Shampa IJ, Sumi SJ and Islam O. (2018). Bioactivities of leaf and root extract of Ceriscoids turgida (Roxb.). Oriental Pharmacy and Experimental Medicine. 18(2):15965. Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 60 Chapter 8 The chemistry of a borrowed life: bioactive potential of the hemi-parasitic herb Striga densiflora (Benth.) Benth. (Orobanchaceae) Sugimani Marndi1, Sanjeet Kumar1, Sweta Mishra1, Subhalakshmi Rout1, Brajesh Kumar Sahu2 and Sumitra Jethy1* 1Ambika Prasad Research Foundation, Odisha, India 2Department of Botany, P.M. College of Excellence, Government College Vidisha, Madhya Pradesh, India *Email-Id:[email protected] DOI: https://doi.org/10.5281/zenodo.17499463 ABSTRACT In the quiet scrublands of Chandaka, Odisha, Striga densiflora thrives as a seemingly ordinary hemi-parasitic plant, yet within its delicate structure lays a reservoir of unexplored chemistry. This study sought to uncover its hidden bioactive potential through a comparative solvent extraction approach. Whole plants were gently washed, shade dried and macerated in solvents of varying polarity to observe how nature’s chemistry unfolds under different conditions. The non-polar extract revealed no major secondary metabolites. In contrast, the polar extract tested positive for tannins, saponins, phenolics, alkaloids, and reducing sugars. When challenged against Escherichia coli, a common and resilient bacterial strain, the hydroethanolic extract displayed complete inhibition of growth at all tested concentrations. The minimum inhibitory concentration (MIC) was determined to be 12.5 mg/mL. These results suggest that S. densiflora harbors promising antibacterial constituents, its potential veiled in polarity and revealed through thoughtful extraction. This work opens a window to the latent therapeutic promise of the plant and invites deeper exploration into its pharmacological essence. Keywords: Antibacterial, semi parasite, therapeutics INTRODUCTION In nature, every plant plays an important role in keeping the environment balanced. While trees and shrubs are easily noticed, small herbs also help in Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 61 maintaining the ecosystem by supporting soil health, providing food, and regulating natural cycles (Guo et al., 2023). Among these herbs, there is a small and special group belonging from the parasitic type. These plants are often considered harmful because they depend on other plants, called hosts, to survive. However, parasitic plants are also examples of how nature evolves in unique ways to adapt and survive in difficult conditions (Bouwmeester et al., 2021). Parasitic plants are mainly divided into two types, holoparasites and hemiparasites (Ishida and Costa, 2024). Holoparasites are completely dependent on their host plants as they cannot produce their own food due to the absence of chlorophyll. On the other hand, hemi-parasitic plants are partly independent. They attach to host plants for water and minerals but can still make their own food through photosynthesis. This makes them more adaptable and resilient in changing environments. Striga densiflora (Figure 1) is one such hemi-parasitic herb which is normally found in seasonally dry tropical regions (POWO, 2025). Some of its reported habitats include upper Gangetic, Karnataka, Tamil Nadu Plains, Odisha and Deccan Peninsula (Lyngdoh et al., 2023). It can grow in harsh conditions using its root as the mode of parasitism (Lyngdoh et al., 2023) and adjust well to environmental changes. Although it is often ignored as a weed, it may contain valuable chemical compounds that have medicinal importance because the members of the Orobanchaceae family, to which S. densiflora belongs, are well known not only for their parasitic lifestyle but also for their significance in traditional medicine. Despite their dependence on host plants, several species in this family are valued for their healing properties. For instance, as reported by a survey (Jaiswal et al., 2021), Aeginetia indica stems are used to relieve coughs, while Buchnera hispida leaves are ground into a paste to treat scabies and eczema. The flowers of Centranthera indica are used as a diuretic, and Orobanche cernua stems are applied to reduce swelling. Likewise, Striga asiatica flower infusions are traditionally used to eliminate stomach parasites. These examples show that even parasitic plants, often viewed as harmful, possess rich medicinal potential. An important indication to how these plants are used as effective folk medicinal practices may link to the fact that, hemiparasitic plants interact closely with other species and face environmental stress, they often produce different secondary metabolites or natural Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 62 chemicals that help them defend and survive (Behera et al.,2023). And, many of these compounds are known to show antibacterial and other health related properties. But, despite its presence in many parts of India, Striga densiflora has not been studied much for its chemical or medicinal properties. This study aims to fill that gap by examining its phytochemical composition and testing its antibacterial activity. By understanding how this hemi-parasitic plant produces and uses these compounds, we can explore its potential as a natural source of antimicrobial agents. Figure 1: Striga densiflora in its habitat METHODOLOGY Plant Collection and Identification: Whole plants of Striga densiflora (Figure 1) were collected from the Chandaka village of Khordha, Odisha, India. The plant species was taxonomically verified using relevant published floras and identification keys (Kumar et al., 2018; Balkrishna et al., 2018). Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 63 Figure 2: Plant parts of S. densiflora Preparation of Plant Material: Collected plants (Figure 2) were rinsed thoroughly under running tap water to remove adhering soil and surface impurities. The cleaned samples were shade dried briefly to eliminate excess surface moisture while retaining volatile constituents. The partially dried plants were then coarsely chopped (Figure 3) and subjected directly to maceration (Devi et al., 2023). Extraction of Bioactive Compounds: Extraction was performed through cold maceration using solvents of varying polarity to ensure comprehensive recovery of phytoconstituents following standard procedures (Devi et al., Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 70 Chapter 9 Climbing towards cure: phytochemical and antibacterial exploration of Gouania leptostachya DC. (Rhamnaceae) Bhagwati Prashad Sharma1, Sanjeet Kumar2, Sweta Mishra2, Sugimani Marndi2, Subhalakshmi Rout2 and Sumitra Jethy2* 1Department of Botany, Sidharth Government College, Nadaun, Himachal Pradesh, India 2Ambika Prasad Research Foundation, Odisha, India *Email-Id: [email protected] DOI: https://doi.org/10.5281/zenodo.17510053 ABSTRACT Lianas being famous for their entwining vines but less studied in light of their pharmacological potential. Gouania leptostachya (Rhamnaceae) is one such liana known for its traditional medicinal uses across tropical regions of Asia. Present study explores the phytochemical composition and antibacterial potential of its leaf extracts. The phytochemical screening showed that the hydroethanolic extract was richer in secondary metabolites, including tannins, saponins, phenolic compounds, and reducing sugars, while the non-polar extract contained only reducing sugars. When tested against Escherichia coli to validate its folk medicinal practices, the hydroethanolic extract exhibited notable antibacterial activity, with a minimum inhibitory concentration (MIC) of 12.5 mg/mL. Observations suggested that the polar extract of G. leptostachya leaves holds promising antibacterial properties, possibly linked to the presence of phenolic and tannin compounds. This study provides a scientific basis for the plant’s traditional use and supports its potential as a natural source of bioactive compounds. Keywords: Bioactivity, folk medicine, liana INTRODUCTION In tropical forests, lianas (long-stemmed woody vines) form an important part of the forest structure. These plants grow from the ground and climb trees or other supports using tendrils, hooked spines, or climbing roots to Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 71 reach sunlight in the upper canopy (Rowe, 2018). Unlike epiphytes that live on other plants, lianas stay rooted in the soil while extending high into the forest layers (Ceballos et al., 2025). Their presence often reflects the health and maturity of a forest ecosystem. Although they compete with trees for light, water, and nutrients, and sometimes slow their growth by adding weight, lianas also play vital ecological roles (De Deurwaerder et al., 2024). They connect tree canopies, creating natural bridges that allow animals to move easily through the forest, and influence forest regeneration and diversity (Schnitzer and Bongers, 2002). In addition to their ecological functions, many lianas have cultural and medicinal importance (Devarakonda et al., 2022). Communities living in tropical regions have long used them in traditional healing, food, and daily practices (Dunget al., 2015). Thus, lianas are not only a structural element of forests but also a source of knowledge and health for people who live close to them. Gouania leptostachya (Figure 1) in that context was studied for correlating its ethnomedicinal practices with today’s modern medicine. Figure 1: Leaves and inflorescence of Gouania leptostachya Being a native species in the Indian subcontinent and Western Malaysia, this Slender Spiked Gouania belongs to family Rhamnaceae. It typically grows in wet tropical biomes and has ethnobotanical relevance which makes it an integral but lesser-known part of the folk medicinal practices (Gumisiriza et al., 2024). Its leaves, stems, and roots are prepared as poultices, extracts, decoctions, herbal baths, and smoke inhalations for the management of ailments including sores, wounds, gastrointestinal disorders, postpartum complications, inflammatory diseases, and fever Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 72 (Table 1). Thus, its potential pharmacological relevance has been assessed in this study to support its traditional therapeutic reputation. Table 1: Traditional ethnomedicinal uses of G. leptostachya Plant parts used Mode of use Ethnomedicinal applications Associated communities/ regions Source(s) Leaves Poultice applied externally/ topically Treatment of eczema (local name: Raktapitchali) India (Odisha, Jharkhand, Uttarakhand, West Bengal) Devarakonda et al., (2022); Pradhan and Badola, (2008); Uprety et al., (2016) Leaf Extract applied externally Treatment of wounds India (Himachal Pradesh) Sharma et al., (2024) Leaves and stems Decoction or cold infusion taken orally Relief from stomachache and diarrhea Indonesia Gumisiriza et al., (2024) Whole plant (mainly leaves) Herbal bath formula and food supplement Postpartum care, treatment of convulsions in newborns, numbness, fainting, antispasmodic conditions and general inflammation Mien tribe, Northern Thailand, Vietnam, Bhutan, Nepal Panyaphu et al., (2011); Dung et al., (2015); Gumisiriza et al., (2024) Roots Roots burned; smoke inhaled Treatment of fever Rural communities of North Andaman, India Prasad et al., (2008) Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 73 METHODOLOGY Leaves of Gouania leptostachya (Figure 2) were collected from Chandaka Khordha, Odisha, India, and identified by the authors. The collected samples were properly tagged and placed in cotton bags to prevent contamination during transport. Figure 2: Leaves of G. leptostachya In the laboratory, the leaves were washed thoroughly under running tap water followed by distilled water to remove adhering dust, debris, and surface contaminants. Cleaned leaves were shade dried at ambient temperature (25 ± 2 °C) for 7-10 days to preserve thermolabile phytoconstituents and then finely powdered using a mechanical grinder. The powdered material was subjected to solvent extraction by the maceration method (Devi et al., 2025) using two solvents of different polarity, n-hexane and hydro-ethanol (1:1), to ensure a broad spectrum of phytochemical recovery. The mixtures were agitated intermittently for 48 hours and filtered through Whatman No. 1 filter paper, after which the filtrates were concentrated at room temperature to evaporate residual solvent. The obtained crude extracts were stored in sterile glass vials under Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 74 refrigeration (4°C) until further analysis. Preliminary qualitative phytochemical screening was performed on both extracts following standard procedures to detect the presence of major classes of secondary metabolites such as tannins, flavonoids, terpenoids, reducing sugars, saponins, alkaloids, carbonyl compounds and phenolic compounds. The antibacterial potential of the extracts was subsequently evaluated against Escherichia coli (MTCC 45). The minimum inhibitory concentration (MIC) was determined by the broth microdilution method following standard protocols with minor modifications (Jena et al., 2025) to assess the concentration-dependent antibacterial efficacy of the extracts. RESULTS AND DISCUSSION The qualitative phytochemical screening of Gouania leptostachya leaf extracts revealed the presence of key secondary metabolites that varied according to solvent polarity (Table 2). The non-polar n-hexane extract detected reducing sugars (Figure 3), indicating a limited phytochemical yield. Meanwhile, the polar hydroethanolic extract exhibited a richer phytochemical profile, containing tannins, saponins, phenolic compounds, and reducing sugars (Figure 4). This suggests that polar solvents are more efficient in extracting diverse bioactive compounds from G. leptostachya leaves, reflecting their higher phytochemical richness. The antibacterial activity of its hydroethanolic leaf extract was tested against E. coli (MTCC 45). The extract effectively inhibited bacterial growth at all tested concentrations, with the minimum inhibitory concentration (MIC) recorded at 12.5 mg/mL (Table 3). The results were consistent across all replicates. Quality control checks verified sterility in the nutrient broth, viability of the microbial culture, and complete inhibition in the antibiotic control. Table 2: Detection of secondary metabolites of G. leptostachya leaf extract Polarity Solvent extract Detected secondary metabolites Phytochemical richness Non-polar n-Hexane Reducing sugars Polar Hydroethanolic Tannins, Saponins, Phenolic compounds and Reducing sugars ( = low, = high) Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 75 Figure 3: Detection of secondary metabolites in n-hexane extract Figure 4: Detection of secondary metabolites in hydroethanolic extract Table 3: Estimation of Minimum Inhibitory Concentration (MIC) Sample Extract Concentration (mg/mL) Result Replicates (n) QC note G. leptostachya (Leaves) Hydroethanolic 100 NG 3 consistent 50 NG 3 consistent 25 NG 3 consistent 12.5 (MIC) NG 3 consistent Nutrient broth - - NG 3 sterility Microbial broth - - G 3 culture viable Antibiotic - - NG 3 positive control (G= Growth, NG= No growth QC = Quality control) Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 76 CONCLUSION Gouania leptostachya is a plant with valuable bioactive potential, particularly evident in its hydroethanolic leaf extract, as observed in the experimental study above. The presented results align with its long-standing traditional uses. While the current findings are preliminary, they open the scope for further research on isolating specific compounds and understanding their mechanisms of action. As a forest liana with deep ethnobotanical roots, G. leptostachya continues to prove how traditional knowledge and modern science can complement each other in exploring nature’s pharmacy. REFERENCES Ceballos SJ, Aráoz E and Rojas TN. (2025). Exploring co-occurrence patterns to understand epiphyte-liana interactions. Plants. 14(1): 140.doi: 10.3390/plants14010140 De Deurwaerder HPT, Detto M, Visser MD, Schnitzer S and Pacala SW. (2024). Linking physiology, epidemiology, and demography: understanding how lianas outcompete trees in a changing world. Proceedings of the National Academy of Sciences of the United States of America. 121(34): e2319487121.doi: 10.1073/pnas.2319487121 Devarakonda R, Mukherjee S, Arya A, Sharma A, Marndi S and Kumar S. (2022). Some common medicinal lianas of India. In: MedicoBiowealth of India, VolumeVI. Ambika Prasad Research Foundation, Odisha, India. Devi RS, Satapathy KB and Kumar S. (2023). Validation of tribal claims for formulation of future drugs through evaluation of ethnopharmacological values of Ludwigia adscendens. Medicinal Plants. 15(4): 691-697. Dung TTM, Lee J, Kim E, Yoo BC, Ha VT, Kim Y, Yoon DH, Hong S, Baek KS, Sung NY, Kim TW, Kim JH and Cho JY. (2015). Anti‐inflammatory activities of Gouania leptostachya methanol extract and its constituent resveratrol. Phytotherapy Research. 29(3): 381-392. Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 77 Gumisiriza H, Olet EA, Mwikali L, Akatuhebwa R, Kembabazi O, Omara T and Lejju JB. (2024). Ethnopharmacology, phytochemistry, pharmacology and toxicity of the genus Gouania. Heliyon. 10(24): e40933. doi: 10.1016/j.heliyon. 2024.e40933 Jena N, Vimala, Singh B, Patra A, Sharma BP, Hossain E and Kumar S. (2025). Methods for ethnobotanical data collection, phytochemistry, antioxidant, anthelmintic, and antimicrobial activities for pharmacological evaluation of medicinal plants. Journal of Biodiversity and Conservation. 9(2): 87-107. Rowe N. (2018). Lianas. Current Biology. 28(6): R249-R252. Panyaphu K, On TV, Sirisa-ard P, Srisa-nga P, ChansaKaow S and Nathakarnkitkul S. (2011). Medicinal plants of the Mien (Yao) in Northern Thailand and their potential value in the primary healthcare of postpartum women. Journal of Ethnopharmacology. 135(2): 226-237. Pradhan BK and Badola HK. (2008). Ethnomedicinal plant use by Lepcha tribe of Dzongu valley, bordering Khangchendzonga Biosphere Reserve, in north Sikkim, India. Journal of Ethnobiology and Ethnomedicine. 4:22. doi: 10.1186/1746-4269-4-22 Prasad PRC, Reddy CS, Raza SH and Dutt CBS. (2008). Folklore medicinal plants of North Andaman Islands, India. Fitoterapia. 79(6): 458-464. Schnitzer SA and Bongers F. (2002). The ecology of lianas and their role in forests. Trends in Ecology & Evolution. 17(5): 223-230. Sharma BP, Sharma AJ and Sharma A. (2024) An ethnomedicinal study on climbers and lianas of Changar region in district Kangra of Himachal Pradesh, India. Journal of Non-Timber Forest Products. 31(2): 139-149. Uprety Y, Poudel RC, Gurung J, Chettri N and Chaudhary RP. (2016). Traditional use and management of NTFPs in Kangchenjunga Landscape: implications for conservation and livelihoods. Journal of Ethnobiology and Ethnomedicine. 12:19. doi: 10.1186/s13002-0160089-8 Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 78 Chapter 10 Natural antibacterial potential of Grewia rhamnifolia Roth (Malvaceae): insights from phytochemical analysis Sumitra Jethy*and Sanjeet Kumar Ambika Prasad Research Foundation, Odisha, India *Email-Id: sumi[email protected] DOI: https://doi.org/10.5281/zenodo.17519659 ABSTRACT Antibiotic resistance has become a major global problem, leading researchers to explore medicinal plants as natural antibacterial agents. This experiment focused on the fruits of Grewia rhamnifolia, a wild plant belonging to the Malvaceae family known for its therapeutic properties. Fruit extracts were prepared using the Soxhlet extraction method with contrasting solvents and tested for their chemical composition and antibacterial activity. The hydroethanolic extract showed the presence of several important compounds such as tannins, phenolics, alkaloids, and reducing sugars, suggesting it was the most effective solvent for extracting active ingredients. The antibacterial test against Escherichia coli showed that the extract could stop bacterial growth at a concentration of 100 mg/mL. These results indicate that G. rhamnifolia fruits contain useful natural compounds with potential antibacterial properties. This study supports the traditional use of this plant and highlights its possible role in developing new plant-based medicines. Keywords: Ethnomedicine, antibiotic resistance, plant-based therapeutics INTRODUCTION Today, the search for natural antibacterial agents has gained considerable global attention due to the crisis of increasing antibiotic resistance (Angelini, 2024). However, traditional knowledge systems known as modern-day ethnobotany and ethnomedicine have long recognized the therapeutic potential of plants, providing a solid foundation for a new era of drug discovery (McClatchey et al., 2009). Worldwide, numerous indigenous communities have historically relied on medicinal plants to treat infectious diseases, guided by generations of experiential knowledge from Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 79 traditional healers and folk medicine practitioners. These plant-based remedies constitute a diverse group of bioactive compounds like flavonoids, phenolics, alkaloids, tannins and many more that add to their antibacterial potency (AlSheikh et al., 2020). This brings us to thepretextthat wild and underexplored plants having medicinal uses can be used for developing safer and sustainable natural antibacterial agents. Grewia rhamnifolia (Figure 1) is one such wild climbing shrub found in seasonally dry tropical biomes native to India and Sri Lanka (POWO, 2025). Figure 1: A twig of Grewia rhamnifolia The leaves of this species are ovate-lanceolate with serrulate or crenulate margins, an acuminate apex, and a rounded or obtuse base. They are typically 3-nerved, occasionally exhibiting two additional smaller lateral nerves, with 3-5 secondary veins, and are nearly glabrous. The flowers are large, white to yellowish in colour, and borne in terminal or leaf-opposed umbels containing few blossoms. Floral buds are ovoid, distinctly ribbed, and covered with dense tomentose and hirsute hairs. The gonophore is short, and the style is stout with a four-lobed stigma. The drupe is depressed-globose, hairy, and yellow tomentose, containing several seeds; Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 86 Dharmasoth RD and Rao BG. (2019). Phytochemical and pharmacological review of Grewia tiliaefolia (VAHL). International Journal of Pharmaceutics. 10(9): 39-42. Gupta MK, Sharma PK, Ansari SH and Lagarkha R. (2006). Pharmacognostical evaluation of Grewia asiatica fruits. International Journal of Plant Sciences. 1(2):249-251. Jena N, Vimala, Singh B, Patra A, Sharma BP, Hossain E and Kumar S. (2025). Methods for ethnobotanical data collection, phytochemistry, antioxidant, anthelmintic, and antimicrobial activities for pharmacological evaluation of medicinal plants. Journal of Biodiversity and Conservation. 9(2): 87-107. McClatchey WC, Mahady GB, Bennett BC, Shiels L and Savo V. (2009). Ethnobotany as a pharmacological research tool and recent developments in CNS-active natural products from ethnobotanical sources. Pharmacology and Therapeutics.123(2):239-254. Morton JF. (1987). Fruits of Warm Climates. Julia F Morton, Miami, USA. Plants of the World Online (POWO). (2025). Grewia rhamnifolia Roth. Royal Botanic Gardens Kew, United Kingdom. Rafe MR, Salam R, Hossain MM and Masud MM. (2018). Phytochemical Screening and acetylcholinesterase and butyrylcholinesterase inhibitory and thrombolytic activities of Grewia abutilifolia Vent. and Juss. Leaf. Dhaka University Journal of Pharmaceutical Sciences. 17(1):81-86. Saxena HO and Brahmam M. (1994). The Flora of Orissa, Volume 1. Regional Research Laboratory, Bhubaneswar, Odisha & Orissa Forest Development Corporation Ltd., Bhubaneswar, Odisha, India. Suguna M and Umesha S. (2022). Phytochemical composition, pharmacological properties, and therapeutic activities of genus: Grewia. Journal of Pharmacognosy and Phytochemistry. 11(4): 263272. Takó M, Kerekes EB, Zambrano C, Kotogán A, Papp T, Krisch J and Vágvölgyi C. (2020). Plant phenolics and phenolic-enriched extracts Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 87 as antimicrobial agents against food-contaminating microorganisms. Antioxidants.9(2): 165. doi: 10.3390/antiox9020165. Ullah W, Uddin G and Siddiqui BS. (2012). Ethnic uses, pharmacological and phytochemical profile of genus Grewia. Journal of Asian Natural Products Research. 14(2):186-195. Waliullah GU, Rauf A, Siddiqui BS, Rehman T, Azam S and Qaisar M. (2011). Chemical constituents and biological screening of Grewia optiva Drummond ex Burret whole plant. American-Eurasian Journal of Agricultural and Environmental Sciences. 11(4): 542546. Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 88 Chapter 11 The rising tide of antimicrobial resistance: causes, consequences and potential solutions Bhagwati Prashad Sharma1, Paramita Ray2, Kadambini Das3 and Rajkumari Supriya Devi2* 1Department of Botany, Sidharth Government College, Nadaun, Himachal Pradesh, India 2Ambika Prasad Research Foundation, Odisha, India 3University Department of Botany, Babasaheb Bhimrao Ambedkar Bihar University, Muzaffarpur, Bihar, India *Email-Id:[email protected] DOI: https://doi.org/10.5281/zenodo.17602605 ABSTRACT Antimicrobial resistance (AMR) has emerged as a defining challenge of modern medicine, threatening to reverse decades of therapeutic progress. The phenomenon, driven by the evolutionary adaptability of microorganisms and exacerbated by human misuse of antimicrobial agents, represents a multidimensional crisis that spans biology, health systems, agriculture, economics, and policy. Present study examines the escalating threat of AMR by exploring its underlying causes, far-reaching consequences, and feasible solutions within a global “One Health” framework. The study highlights the ecological, social, and economic repercussions of rising resistance and the emerging recognition of AMR as a silent pandemic. The study also outlines potential solutions, ranging from stewardship programs and regulatory reforms to innovative therapies and public awareness initiatives. The overarching argument emphasises that AMR is not a future threat but an immediate reality requiring urgent, coordinated, and sustained global action. Keywords: AMR, drugs, one health, pathogenic microbes Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 89 INTRODUCTION Antimicrobial resistance refers to the ability of microorganisms to withstand the effects of drugs that were once effective against them. This resistance develops through natural selection, genetic mutation, or horizontal gene transfer, enabling pathogens to survive and multiply despite exposure to antimicrobial agents (Salam et al., 2023; Figure 1). In practical terms, AMR renders many common infections increasingly difficult or impossible to treat, turning minor ailments into life-threatening conditions (Muteeb et al., 2023). The growing prevalence of resistant pathogens has created a major global health concern that transcends national borders and healthcare systems. The discovery of penicillin by Alexander Fleming in 1928 revolutionised modern medicine, heralding the antibiotic era (Lobanovska and Pilla, 2017). However, even in his Nobel lecture, Fleming warned of the potential misuse of antibiotics, leading to resistance a prophecy now realised (Rosenblatt-Farrell, 2009). The decades following World War II witnessed the discovery of numerous antibiotic classes, resulting in a golden age of antimicrobial therapy (Adedeji, 2016).Yet, excessive use in humans, animals, and agriculture, coupled with a decline in the development of new antibiotics, has led to a critical imbalance. The same drugs that once symbolised scientific triumph are now losing their potency, as resistant pathogens outpace pharmaceutical innovation (Brüssow, 2024). Unlike acute health crises such as COVID-19, antimicrobial resistance spreads quietly, without public panic or immediate visibility. Its cumulative impact, however, is profound. Resistant infections cause prolonged illnesses, increased hospital stays, and elevated mortality rates (Adebisi et al., 2021). The inability to treat routine infections jeopardises modern medical procedures such as surgeries, chemotherapy, and organ transplants, all of which depend on effective antimicrobial prophylaxis (Chinemerem et al., 2022). The silent spread of resistance genes across borders underscores that AMR is not confined to any single region or species; it is a planetary health issue that challenges global sustainability. In recent years, the burden of antimicrobial resistance has grown exponentially (Salam et al., 2023). Multidrug-resistant tuberculosis, methicillin-resistant Staphylococcus aureus (MRSA), and carbapenem- Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 90 resistant Enterobacteriaceae have become emblematic of the crisis. Estimates suggest that millions of deaths annually are attributable to resistant infections, with the majority occurring in lowand middle-income countries (Marino et al., 2025). Figure 1: Mechanisms of microbial resistance development In many regions, inadequate healthcare infrastructure, poor sanitation, and unregulated access to antibiotics contribute to the rapid spread of AMR. Additionally, international travel and trade facilitate the global dissemination of resistant strains, blurring the boundaries between local and global outbreaks (Endale et al., 2023). The complexity of AMR arises from its deeply interconnected causes. Beyond biological evolution, resistance is influenced by sociocultural practices, economic pressures, and environmental degradation (Larsson and Flach, 2022). The unregulated use of antibiotics in livestock and aquaculture fosters resistance in animal populations, which can transfer to humans through food chains or environmental contamination (Manyi-Loh et al., 2018). Furthermore, pharmaceutical manufacturing waste containing antibiotic residues further pollutes our water systems, creating environments that promote the growth of resistant bacteria. AMR thus epitomizes a systems-level problem that demands a holistic understanding and coordinated global action (Kotwani et al., 2021). Traditional approaches that focus solely on medical interventions are insufficient. Addressing the AMR crisis requires a “One Health” perspective, which acknowledges the intrinsic links between human, animal, and environmental health (Danasekaran, 2024). To effectively combat resistance development, integrated surveillance, rational drug use, Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 91 improvements in sanitation, and educational campaigns must operate in synergy. Ultimately, tackling the problem of AMR involves not just microbiology but also issues of governance, awareness, and equity as it is about microbiology. CAUSES AND DRIVERS OF ANTIMICROBIAL RESISTANCE Antimicrobial resistance (AMR) does not arise from a single source but from a convergence of biological adaptation, human behaviour, agricultural practice, and policy shortcomings. Its drivers operate at multiple scales,from molecular genetics within microbes to global trade and governance failures. Understanding these interconnected causes is essential for designing meaningful interventions (Ferraz, 2024). Microorganisms possess remarkable genetic plasticity that enables them to adapt rapidly to hostile environments. When exposed to antimicrobial agents, selective pressure favours the survival of resistant variants, which then proliferate. Resistance can emerge through several mechanisms, each conferring unique adaptive advantages.Random mutations in microbial DNA can alter drug targets, metabolic pathways, or membrane permeability. These mutations may reduce a drug’s binding affinity, deactivate its enzymatic target, or prevent its entry into the cell. In bacterial populations that reproduce quickly, even a single advantageous mutation can dominate within hours under antibiotic pressure (Salam et al., 2023).Perhaps the most powerful driver of AMR is the exchange of genetic material between microorganisms. Through conjugation, transformation, or transduction, bacteria can acquire resistance genes carried on plasmids, transposons, or integrons.This means that resistance developed in one speciessuch as environmental Pseudomonas,can transfer to a human pathogen like Escherichia coli. The “resistome,” a collective gene pool of resistance determinants, circulates freely across microbial communities in soil, water, and the human microbiome (Iskandar et al., 2022).Many microbes form biofilms,protective matrices of polysaccharides and proteins that shield bacterial cells from antibiotics. Within biofilms, slow-growing or dormant “persister” cells evade antimicrobial action, only to repopulate once treatment ceases. This physiological tolerance differs from genetic resistance but contributes significantly to treatment failure in chronic Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 92 infections (Iskandar et al., 2022). Exposure to non-antibiotic compounds such as heavy metals, disinfectants, or biocides can co-select for resistant strains. Likewise, a single resistance mechanism,such as efflux pumps,can expel multiple drug classes simultaneously, leading to cross-resistance. These intertwined pathways illustrate why resistance emerges even in environments not directly exposed to antibiotics (Endale et al., 2023). Human and clinical factors: Human medical practices are the primary contributors to AMR. The overuse and misuse of antibiotics in healthcare settings accelerate the development and spread of resistance. Antibiotics are often prescribed for viral infections, such as colds or influenza, where they offer no benefit. In many regions, patients can purchase antibiotics without a prescription, leading to incomplete or inappropriate treatment courses. Every unnecessary exposure to antibiotics strengthens selective pressure,raising the likelihood that resistant mutants will survive.In lowresource settings, the lack of rapid and accurate diagnostic tools forces clinicians to rely on empirical treatment(Castro-Sanchez et al., 2016). As a precaution, broad-spectrum antibiotics are administeredeven when the cause of infectionis uncertain. This indiscriminate use eliminates susceptible microbes while leaving resistant populations to thrive.Healthcare facilities, particularly intensive care units, serve as hubs for resistant pathogens. Inadequate infection prevention measures, poor hand hygiene, contaminated equipment, and overcrowded wardsfacilitate cross-infections.Resistant strains such as Methicillin-resistant Staphylococcus aureus (MRSA) and Clostridioides difficile often spread within hospitals, compounding patient morbidity and healthcare costs.Patients who discontinue their treatment prematurely, once their symptoms subside, allow partially resistant microbes to survive and multiply (Castro-Sanchez et al., 2016). Similarly, sub-therapeutic dosing or counterfeit drugs with insufficient active ingredient content create ideal conditions for the selection of resistance. The movement of people across borders further spreads resistant organisms internationally. A patient treated with antibiotics abroad may return home carrying multi-resistant strains, effectively globalising local outbreaks. The interconnected nature of healthcare systems ensures that resistance knows no geographic boundaries (Endale et al., 2023). Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 93 Agricultural and environmental factors: Antimicrobials have become integral to modern food production, yet their indiscriminate use outside human medicine profoundly influences resistance evolution (Li et al., 2023).Antibiotics are widely administered to animals not only for therapeutic purposes but also as growth promoters and prophylactics. Continuous low-dose exposure in feed creates optimal conditions for resistance selection within animal microbiota (Laborda et al., 2022). Resistant bacteria or resistance genes can then transfer to humans via direct contact, contaminated meat, or environmental runoff.Fish farming operations often rely on antibiotics to prevent disease outbreaks in dense populations. Similarly, certain crops are treated with antibacterial agents to control plant pathogens. These practices introduce substantial amounts of antimicrobial residues into aquatic and soil environments, fostering resistant microbial communities (Zeng et al., 2025).Effluents from drug manufacturing, hospitals, and municipal waste streams carry unmetabolized antibiotics into rivers and soils. In these habitats, sub-inhibitory concentrations of antimicrobials exert chronic selection pressure, enriching for resistant bacteria even in the absence of disease (Laborda et al., 2022). Such environmental reservoirs act as breeding grounds where resistance genes can recombine and later re-enter human or animal populations.Wild animals exposed to human waste or agricultural effluents can acquire and disseminate resistant bacteria through migratory routes. This ecological connectivity underscores that AMR extends beyond anthropogenic boundariesit permeates entire ecosystems (Laborda et al., 2022). Socioeconomic and policy drivers: Hidden beneath the biological and behavioral layers of AMR lie deep structural inequities and policy failures that perpetuate resistance on a global scale.In some regions, antibiotics are scarce, leading to under-treatment, while in others, they are easily accessible over the counter. The lack of strong regulatory frameworks allows counterfeit or substandard drugs to circulate, increasing the risk of resistance.Economic constraints often compel patients to purchase incomplete doses, resulting in incomplete treatment.To effectively contain AMR, it is essential to gather data on antibiotic use and resistance patterns. Many countries lack functional surveillance systems and inconsistently enforce regulations. Without evidence-based policies, antibiotics continue Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 94 to be misused across different sectors, and resistant outbreaks often go undetected until they escalate.Additionally, the antibiotic development pipeline has stagnated because new drugs offer low commercial returns compared to treatments for chronic-disease. As a result, pharmaceutical complete are reluctant to invest in costly research for medicines intended for short-term use,especially when they are subject to stewardship guidelines. This economic disincentive has resulted in a global reliance on aging classes of antibiotics.The flow of goods, animals, and people facilitates international spread of resistant strains. Food exports that are contaminated with resistant bacteria or resistance genes can create new ecological niches in far-off markets. At the same time, global trade agreements often overlook public-health safeguards, prioritizing economic interests over the responsible use of antimicrobials.Social perceptions about antibiotics influence their consumption. In many societies, antibiotics are perceived as quick fixes for any illness, which reinforce their demand and misuse (Capuozzo et al., 2024). A lack of undersatnding about resistance mechanisms perpetuates behaviors that inadvertently fuel the crisis. Therefore, education and cultural change are therefore as vital as scientific discovery.These factors are interconnected within a complex web of causality.Biological evolution is influenced by human behavior, while human misuse is shaped by socioeconomic pressures. Environmental contamination provides the backdrop for the ongoing circulation of resistance. This interplay blurs the boundaries between clinical, agricultural, and ecological issues. No single intervention, whether scientific or regulatory,can succeed in isolation. An effective strategy must address the underlying network of interactions that sustain antibiotic resistance (Singer et al., 2025). CONSEQUENCES AND IMPACTS OF ANTIMICROBIAL RESISTANCE Antimicrobial resistance (AMR) has become one of the gravest global health challenges, with repercussions extending far beyond clinical medicine. Its impacts are multifaceted, spanning healthcare systems, economies, ecosystems, and societies.As infections resistant to treatment grow in prevalence, the world faces a scenario where oncemanageable Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 95 diseases could again become fatal. Therefore, the consequences of AMR are not just a biomedical issue; they represent a significant threat to human development, environmental stability, and social equity (de Kraker, 2023; Figure 2). Clinical and public health burden: The most immediate consequence of antimicrobial resistance is the increasing difficulty in treating infectious diseases. Infections that were once easily curable now require longer, more complex, and often less effective treatment regimens. Patients infected with resistant organisms experience prolonged illness, recurrent infections, and higher rates of complications. Common bacterial diseases, such as pneumonia, urinary tract infections, and sepsis, have become progressively harder to manage, especially in hospital settings. Multidrug-resistant pathogenssuch as Staphylococcus aureus, Klebsiella pneumoniae, Mycobacterium tuberculosis, and Neisseria gonorrhoeae are causing global concern.These “superbugs” have developed resistance to multiple classes of antibiotics, limiting available treatment options. In extreme cases, infections become entirely untreatable. The emergence of pan-resistant strains indicates a potential collapse of modern infectious disease control (Prestinaciet al., 2015). Figure 2: Conceptual framework of antimicrobial resistance drivers and consequences Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 102 and data-sharing platforms is improving real-time monitoring and outbreak detection (Karpet al., 2017). Data gaps and limitations: Despite some progress, global data on AMR remain fragmented. Many low-income countries lack the infrastructure needed for routine microbial culture or susceptibility testing. Even when data are available, they often do not represent infections at the rural or community level. Additionally, there is limited integration of veterinary and environmental data, which hinders comprehensive One Health analysis. Bridging these gaps is essential for achieving global equity and ensuring scientific accuracy (Nazir et al., 2025). Lessons from the COVID-19 pandemic: The COVID-19 pandemic has provided important insights into the interconnectedness of global health crises. It highlighted both the strengths and weaknesses of international health coordination,lessons that are directly applicable to AMR.During the pandemic, the widespread use of antibiotics in COVID-19 patients, often administered without confirmed bacterial co-infection, led to increased resistance rates in several countries. The crisis also exposed vulnerabilities in supply chains, surveillance, and public communication,which are similar to those encountered in managing AMR. However, it also accelerated innovations such as genomic surveillance, international data sharing, and investment in diagnostics—all of which can strengthen AMR control.The pandemic reinforced the importance of global solidarity, rapid response mechanisms, and transparent data sharing. Just as viruses spread globally, so do resistant bacteria and genes. The internationalmobilisation against COVID-19 provides a template for the coordinated, cross-sectoral action required to combat AMR, though this effort must be sustained beyond acute crisis moments (Khoo and Lantos, 2020). POTENTIAL SOLUTIONS AND FUTURE DIRECTIONS The growing tide of antimicrobial resistance (AMR) demands an integrated, multi-sector approach that looks to the future. Although this crisis has developed over several decades, it is not impossible to tackle. The solution involves a combination of responsible antimicrobial use, scientific innovation, improved governance, and global cooperation within a unified “One Health” framework. The next section examines the key strategies for Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 103 managing and reducing AMR, emphasising both immediate actions and long-term plans essential for sustainable control (Puri et al., 2025). Antimicrobial stewardship programs: The core principle of AMR containment is stewardship, which involves the responsible management of existing antimicrobials to maintain their effectiveness. Antimicrobial stewardship programs (ASPs) are designed to ensure that the correct drug is administered at the appropriate dose and for the right duration, only when necessary. By optimisingthe use of antimicrobials, these programs help minimise pressure on microbes and reduce the development of resistance (Bankar et al., 2022). Clinical stewardship initiatives: Hospitals and healthcare institutions play a pivotal role in antimicrobial stewardship programs (ASPs). Structured ASPs promote evidence-based prescribing through established protocols, formularies, and review systems. Key strategies for effective stewardship include: 1. Implementing diagnostic stewardship to confirm bacterial infections before prescribing antibiotics. 2. Conducting regular audits and providing feedback on antibiotic use patterns. 3. Restricting access to high-risk broad-spectrum antibiotics. 4. Promoting infection prevention measures, such as hand hygiene and environmental sanitation. In high-income settings, electronic prescribing systems and decisionsupport tools assist physicians in adhering to guidelines. In resource-limited contexts, stewardship efforts may focus more on training healthcare workers and ensuring access to quality drugs (Tinker et al., 2021). Community and pharmacy-pased interventions: Stewardship must also extend beyond hospitals. Community-level strategies, especially in outpatient clinics and pharmacies,are crucial since most antibiotics are used outside of hospital settings. Educational campaigns, pharmacist counseling, and public awareness initiatives can helpreduce self-medication and overthe-counter sales of antibiotics. Empowering pharmacists to act as Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 104 gatekeepers in antibiotic dispensing has proven effective in several regions (Zerbinato et al., 2025). Behavioral and cultural change: Sustainable stewardship requires a shift in societal attitudes toward antibiotics. To reduce the common belief that every illness needs medication, we need long-term public education efforts. Incorporating awareness of antimicrobial resistance (AMR) into school curricula, mass media, and health literacy programs can foster responsible antibioticsuse culture. Therefore, stewardship is as much a social movement as it is a medical initiative (Shutt et al., 2025). Novel therapeutics and technologies: Stewardship can help delay the development of resistance, but innovation is essential for keeping pace with it. The global shortage of new antimicrobials is due to both scientific challenges and economic disincentives. However, emerging technologies and unconventional approaches offer hope for a new era of strategies to combat infections.There is renewed urgency in the search for new classes of antibiotic. Advances in genomics, synthetic biology, and high-throughput screening have accelerated the identification of novel compounds. Exploring the soil microbiome and utilizing AI-assisted drug discovery are uncovering previously untapped natural and synthetic molecules. Additionally, structural modifications of existing antibiotics,such as βlactamase inhibitors,can restore activity against resistant strains.Bacteriophagesrepresent a promising alternative to traditional antibiotics. Phage therapy offers specificity, targeting only pathogenic bacteria without disturbing normal microbiota. With advances in genetic engineering, these phages can be tailored to bypass bacterial defense mechanisms. Clinical interest in phage therapy has resurged, especially for treating multi-resistant infections where antibiotics fail.Antimicrobial peptides (AMPs),which are naturally occurring molecules found in animals and plants, present another frontier in drug development. They disrupt microbial membranes, making it harder for resistance to develop. Additionally, host-directed therapies that enhaance immune defenses, rather than targeting microbes directly, may reduce selective pressure and extend the efficacy of existing drugs (Elshobary et al., 2025). Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 105 CRISPR-Cas and genetic tools: Gene-editing technologies, such as CRISPR-Cas systems, can specifically target and disable resistance genes in bacterial populations. These precision tools have the potenmtial to restore bacterial susceptibility or prevent the horizontal transfer of resistance traits. Although still in experimental stages, CRISPR-based antimicrobials represent a significant advancement in the fight against AMR (Tao et al., 2022). Probiotics and microbiome restoration: Maintaining a healthy microbiome provides natural resistance against harnful pathogens. Probiotics and fecal microbiota transplantation have shown potential in restoring gut flora that has been disrupted by by antibiotic use. By supporting microbial balance, these approaches may decrase the recurrence of infectionsand help control the spread of antibiotic resistance (Patangia et al., 2022). Nanotechnology and drug delivery innovations: Nanoparticle-based drug delivery systems can enhance the effectiveness of antibiotics by improving target specificity and reducing the necessary dosage. Nanomaterials with intrinsic antimicrobial properties, such as silver or graphene-based compounds, can also complement conventional therapies. These technologies connect material science and medicine, offering versatile tools to combat resistant pathogens (Parvin et al., 2025). POLICY, GOVERNANCE, AND EDUCATION Effective containment of antimicrobial resistance (AMR) relies on strong policy frameworks that regulate the production, sale, and use of antibiotics. Governments need to establish strict licensing systems for antimicrobials, monitor sales data, and impose penalties for illegal distribution. It is equally important to strengthen drug quality control laboratories and take action against counterfeit medicines. Since resistance knows no borders, global governance mechanisms must ensure accountability and equity. International agreements similar to climate treaties, could formalize national commitments to control AMR. These frameworks should strike a balance between access to and stewardship of antibiotics,ensuring that lifesaving medications remain available to everyone while preventing misuse. Additionally, governments and international donors must realign incentives Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 106 for the research and development of new antibiotics. Push mechanisms, such as grants and public–private partnerships, can help reduce research costs. On the other hand, pull incentiveslike market entry rewards and patent extensionscan make antibiotic innovation financially viable. At the same time, separating profits from sales volume ensures that new antibiotics remain available as last-resort options rather than being marketed excessively. Education is fundamental for long-term control of AMR. It is crucial to train healthcare professionals, veterinarians, and pharmacists in responsible prescribing practices. Additionally, community education campaigns should demystify antibiotics, emphasizing that not all infections require them. Empowering local leaders, schools, and media can turn awareness into action. Policy and governance should be guided by reliable data. Investing in surveillance systems, laboratory networks, and digital reporting tools enhances evidence-based decision-making. Furthermore, data transparency builds public trust and encourages international collaboration (Rogers et al., 2019). RESEARCH AND INNOVATION FOR THE FUTURE Artificial intelligence (AI) and machine learning hold immense potential for predicting resistance trends, designing new molecules, and guiding treatment decisions.AI-driven platforms can analyse large genomic and epidemiological datasets, providing insights into emerging threats before they spread widely.Whole-genome sequencing is transforming the tracking of resistance genes. By identifying transmission chains and evolutionary pathways, genomic tools facilitate targeted interventions. Integrating these technologies into national surveillance programs ensures the vaailability of real-time, actionable intelligence. No single country can address antimicrobial resistance (AMR) in isolation. International research collaborations that pool expertise, data, and funding are vital for accelerating discoveries. These collaborative platforms can standardise methodologies, prevent duplication of efforts, and ensure equitable access to innovations across different regions.However, technical solutions alone are not enough. Understanding cultural attitudes, prescribing behaviors, and public perceptions is crucial for designing interventions that resonate with people. Behavioral science can help inform communication strategies and Plants and Secondary Metabolites, Volume 11; (2025); ISBN: 978-81-989192-9-8 107 encourage populations to use antibiotics more responsibly (Branda and Scarpa, 2024). Pathways to a sustainable AMR future: The path forward must combine innovation with accountability. Stewardship and surveillance provide the foundation, while research and policy provide the necessary tools.Public engagement is essential for ensuring sustainability. Future success depends on integrating these pillars into cohesive global systems that overcomedisciplinary and geographic boundaries.The ultimate goal is not to eliminate microbial resistance,an impossible task,but to restore balance between humans and microorganisms. By promoting responsible use, encouraging robust innovation, and fostering shared global responsibility, humanity can safeguard the effectiveness of antimicrobials for generations to come (Aslam et al., 2024). CONCLUSION Antimicrobial resistance (AMR) has emerged as a major challenge of modern medicine, threatening to reverse decades of progress in scientific and public health. This crisis is complex and is influenced by factors such as microbial evolution, human misuse of antibiotics, reliance on agricultural practices, and neglect of environmental concerns. The rising rates of illness, economic losses, and ecological disruptionhighlight the urgent need for coordinated global action. Tackling AMR requires a comprehensive One Health approach that unites stewardship, innovation, governance, and education across human, animal, and environmental sectors. The future hinges on striking a balance between preserving existing antimicrobials and pursuing new solutions through research, technology, and responsible policies. Only through global solidarity and sustained commitment can we protect the effectiveness of these life-saving drugs and ensure a resilient and sustainable future for generations to come. REFERENCES Adebisi YA, Alaran AJ, Okereke M, Oke GI, Amos OA, Olaoye OC, Oladunjoye I, Olanrewaju AY, Ukor NA and Lucero-Prisno DE. (2021). 3rd. 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Origin of Antibiotics and Antibiotic Resistance, and Their Impacts on Drug Dr. Kugarthi Jayalakshmi,former faculty in the Department of Biochemistry, Sri Padmavathi Mahila Viswavidyalayam, Tirupati–517502, pursued her B.Sc. (2003), B.Ed. (2005), M.Sc. (2007), and Ph.D. (2017) in Biochemistry from Sri Venkateswara University, Tirupati, securing distinction at graduation and first division in postgraduation. She was the topper in SVUCET2005 and secured the first rank in SVURCET-2009. She has been engaged in teaching and research in biochemistry for the last 18 years. Biochemistry, pharmacology and toxicology, and conservation biology of medicinal plants are the key research areas of her interest. She has successfully developed protocols for several medicinal plants, viz., Rhinacanthus nasutus, and has effectively bioprospected to identify bioactive natural products from various medicinal species. She has published 15 research articles in reputed national and international journals, numerous book chapters, authored three books and has one editorial book to her credit. Dr. Preeti Chaturvedi,Professor & Head, Department of Biological Sciences, G.B. Pant University of Agriculture & Technology, Pantnagar. She has completed her B.Sc. (1994), M.Sc. (1996), and Ph.D. (2001) from Kurukshetra University, all in first division with distinction at graduation and post-graduation level. She received the CSIR JRF during 1998–2000. She has been engaged in teaching and research in botany for the last 25 years. Bioprospecting, biodiversity, and conservation biology of medicinal plants and bryophytes are key research areas of her interest. She has successfully developed micropropagation protocols for several high value medicinal plants of Himalaya viz., Aconitum balfourii, Picrorrhiza kurroa,Polygonatum verticillatum,Rheum emodii and Arnebia benthamii. She has bioprospected Himalayan herbs and cryptogams, particularly bryophytes, to identify bioactive natural products. Dr. Preeti has more than 80 research publications in reputed national and international journals and one edited book to her credit. She has successfully completed five research projects. She has supervised twenty-three research students, including 10 Ph. D. and 13 M.Sc. scholars as principal advisor, and more than 20 as co-advisor. Dr. Madhuri Vajha ● Dr. Kugarthi Jayalakshmi ● Dr. Preeti Chaturvedi 'Plants & Secondary Metabolites, Volume 11' offers a comprehensive exploration of ten lesser-known plants, unveiling their botany, phytochemistry, traditional medicinal uses, and potential pharmacological applications, making it an invaluable resource for researchers and botanists. ISBN: 978-81-989192-9-8