A Review on Enhydra Fluctuans Lour: Botanical, Phytochemical, Pharmacological, And Ethnomedicinal Perspectives

Abstract

Enhydra fluctuans Lour., a semi-aquatic herb from the Asteraceae family, is widely recognized in traditional medicine and local diets across South and Southeast Asia. Rich in bioactive compounds such as flavonoids, saponins, tannins, alkaloids, and phenolics, the plant exhibits a broad spectrum of pharmacological activities, including antioxidant, antidiabetic, anti-inflammatory, hepatoprotective, antimicrobial, analgesic, anticancer, and CNS depressant effects. Traditionally used to manage ailments like liver disorders, skin diseases, diabetes, and digestive issues, E. fluctuans is also nutritionally valuable, being high in calcium, iron, vitamin C, beta-carotene, and dietary fiber. Phytochemical and pharmacological studies have utilized Soxhlet extraction, cold maceration, and TLC techniques to isolate active constituents. Though traditionally consumed as a leafy vegetable or in decoctions, modern formulations include capsules, tablets, teas, and topical products. Despite limited commercial availability, growing scientific interest underscores its potential as a therapeutic and functional food source. Despite its therapeutic potential, commercial availability remains limited, primarily due to insufficient large-scale cultivation and standardized processing techniques. E. fluctuans thrives in waterlogged soils under tropical conditions and is propagated both by seeds and vegetative cuttings. Agronomic studies suggest it has a relatively short cultivation cycle, making it suitable for intercropping and wetland farming systems. Toxicological evaluations, including acute toxicity and genotoxicological studies, have indicated a favorable safety profile at traditional dosage levels; however, comprehensive long-term studies are still lacking to confirm its safety for widespread clinical use.

Keywords

Introduction

Enhydra fluctuans Lour., commonly known as water cress, helencha, or jalbrahmi, is a semi-aquatic, herbaceous plant belonging to the family Asteraceae,1 known for its medicinal properties. The plant is grown in tropical and subtropical climates.2 It is widely distributed across South and Southeast Asia, especially in India, Bangladesh, Sri Lanka, and parts of China.1 It is a prostrate, spreading, semi-aquatic annual herbaceous plant with serrated leaves. The use of plants as a medicine was an ancient tradition and is an integral part of the health care system in India.2 This plant thrives in marshy lands, riverbanks, ponds, and rice fields, often forming dense mats over water surfaces due to its creeping, soft-stemmed growth habit.1 In India, there is a wide range of alternative forms of medicine available to the people who do not like to use traditional medicine or who are unable to be cured by traditional medicine. The plant has slightly bitter leaves traditionally used to treat inflammation, skin diseases, and smallpox.2  Enhydra fluctuans is a plant with significant therapeutic value and has been used for centuries in traditional medicine, particularly in remote regions within the Indo-Myanmar biodiversity zone, one of the 25 recognized global biodiversity hotspots.3 Traditionally, E. fluctuans has been valued in Ayurvedic and folk medicine for its therapeutic properties. In India and Bangladesh, it is consumed as a leafy vegetable and used in ethnomedicine for treating various ailments, including nervous disorders, inflammation, liver dysfunction, skin diseases, and hypertension.4 It is also believed to have cooling, laxative, and blood-purifying effects.5 Recent phytochemical investigations have revealed that E. fluctuans contains a rich array of bioactive compounds, including flavonoids, phenols, tannins, saponins, steroids, and alkaloids.5,6 These constituents are believed to contribute to its reported pharmacological activities. Several studies have confirmed its antioxidant, antimicrobial, anti-inflammatory, hepatoprotective, and neuroprotective properties in vitro and in vivo.3,4,7 Despite its widespread traditional usage and growing scientific interest, Enhydra fluctuans remains relatively underexplored. A thorough and systematic evaluation of its phytochemistry, pharmacological effects, and nutritional value may unlock its potential as a functional food and source of novel therapeutic agents.8

2. Plant profile

Fig.: Enhydra fluctuans

Taxonomical Classification

Kingdom	Plantae
Phylum	Mangoliopsida
Class	Magnoliopsida
Order	Asterales
Family	Asteraceae
Genus	Enhydra
Species	Enhydra Fluctuans Loureiro

Synonyms:10

Meyera fluctuans (Lour.), Enhydra anagallis Gardner, and Spreng.

Vernacular Names:4,6

Language	Name
English	Marsh herb, Water cress
Marathi	Nirabela
Hindi	Harkuch, Matsayaakshi, Raktakarabi
Sanskrit	Hilamochika, Jalabramhi
Bengali	Helencha
Telugu	Jalkanda
Gujarati	Jalkand

Common names:

Enhydra fluctuans Lour., known as Indian Marshweed or Raktakarabi, is identified by several regional and local names. Commonly used names include Indian Marshweed, Raktakarabi, Tikhur, Jalkanda, Jalmisri, Jalamanshi, Raktakaravirah, Jalkaravi, and Kanakchampa. These names may differ depending on the region or language, but all refer to the same plant species.11,12

Morphological Features

Enhydra fluctuans is a semi-aquatic, creeping, branched, herbaceous plant that grows in marshy areas and wetlands. It has soft, fleshy stems that root at the nodes, supporting its horizontal spread. Leaves are simple, sessile, linear to oblong, and slightly succulent. The flowers are small, white, and borne in solitary heads in the leaf axils. The plant reproduces both sexually (via seeds) and vegetatively (through stem cuttings or rooting nodes).5,6

Botanical Description:2,13

• Habit: Prostrate, spreading herb that can root at the nodes. It often grows along the edges of water bodies or in marshy areas.

• Stem: Its stems are branching, hairy, rooted at the lower nodes, and grow up to 30 cm or more in length. surfaces.

• Leaves: The leaves are linear-oblong, stalkless, measuring 3 to 5 cm, with blunt or pointed tips, typically truncate bases, and serrated margins.

• Involucral bracts: The involucral bracts are unequal; the outer pair is oval and 1 to 1.2 cm long, while the inner pair is slightly smaller.

• Flowering Heads: Flowering heads are solitary, stalkless, and borne in the leaf axils, measuring less than 1 cm in diameter, excluding the bracts.

• Flowers: The greenish-white or white flowers, usually solitary or in small clusters in the leaf axils, bloom during the warm seasons, and their achenes are enclosed by rigid receptacle scales.

• Fruit: A small achene (a type of dry fruit).

Habitat and Distribution

Enhydra fluctuans L. is a tropical aquatic herbaceous plant that thrives in consistently moist environments. It is commonly found in wetlands, rice paddies, ponds, ditches, and other damp habitats. Well-suited to tropical climates, this species is particularly vulnerable to low temperatures during its early developmental stages. Native to South and Southeast Asia, E. fluctuans is especially widespread throughout India, where it inhabits riverbanks, marshlands, and waterlogged lowlands. Prominent ecosystems supporting its growth include the Sundarbans mangroves of West Bengal, the backwaters and swampy regions of Kerala, and the floodplains and wetlands of Assam. Outside of India, the plant is extensively distributed across neighboring Southeast Asian countries such as Bangladesh, Myanmar, and Thailand. It thrives along major rivers like the Brahmaputra and Ayeyarwady and within various swampy wetland systems. In Sri Lanka, Enhydra fluctuans is found in the coastal zones of Mannar and Jaffna, as well as in the inland wetland areas of Anuradhapura and Polonnaruwa. Its distribution also extends to other parts of Asia, including Nepal, Indonesia, Malaysia, and the Philippines, where it occupies moist lowland areas and upland regions up to elevations of approximately 1,800 meters.14,15

Geographical Distribution

Enhydra fluctuans is a hydrophytic plant commonly found in wet environments such as roadside canals and marshy wastelands, particularly between November and January. It is widely distributed across Bangladesh, China, Malaysia, various parts of Southeast Asia, and tropical regions of Africa. In India, the plant is especially prevalent in Assam and the northeastern states. It is frequently observed in the flora of the Barak Valley and Cachar region of Assam, and is notably dominant in areas such as Majuli Island and Dhemaji district. In Manipur's Thoubal district, Enhydra fluctuans is widely used in traditional medicine to treat various ailments. The Jaintia Hills of Meghalaya and Agartala in Tripura are also known to be rich sources of this plant.16

Traditional Uses

     Enhydra fluctuans is used as an edible plant. The leaves are slightly bitter; they cure inflammation, skin disease, and are good for smallpox. As a traditional medicine, it is commonly used in India for cooling, carminative, tonic, liver-tonic, leprosy, coughs, and especially in hepatopathy. In traditional books, the leaves are used in high blood pressure conditions.14

3. Phytochemical Constituents

Enhydra fluctuans is a rich source of β-carotene, proteins, and a variety of bioactive compounds. These include various chemical constituents such as saponins, alkaloids, flavonoids like apigenin, luteolin, kaempferol, and quercetin. triacontanol (also known as myricyl alcohol) and various phytosterols such as cholesterol, sitosterol, stigmasterol, and stigmasta-5,22,25-triene-3β-ol.12. Phytochemical analyses of Enhydra fluctuans using various solvents (ethanol, methanol, chloroform, aqueous) have revealed the presence of Flavonoids, Phenolic compounds, Alkaloids, Saponins, Steroids, Tannins, Glycosides, Terpenoids, Carbohydrates, Proteins, and amino acids.  These compounds are typically responsible for the plant’s antioxidant, anti-inflammatory, antimicrobial, and hepatoprotective activities.18,19,20. The plant also contains glucosides, diterpenoid acids, along with their isovalerate and angelate derivatives, and sesquiterpene lactones, particularly germacranolides like enhydrin, fluctuanin, and fluctuanidin. Gibberellins A9 and A13 have been isolated from this plant.14,21 Additional compounds include kaurene derivatives, 4-hydroxy farnesyl acetate, and gibberellins. Analysis of the leaf essential oil revealed that myrcene (37%) and limonene (28%) are the major constituents, while minor components (less than 5%) include α-pinene, β-ocimene, camphor, β-caryophyllene, α-humulene, 1-octen-3-ol, linalool, and longiverbenone. Two newly identified components from the leaf oil are cis- and trans-1,2-dihydroperill aldehydes. Methanolic extracts of the plant yield flavonoids, carbohydrates, reducing sugars, phenolics, and tannins.23 Two notable flavonoids, baicalein 7-O-glucoside and baicalein 7-O-diglucoside, have demonstrated cytotoxic, analgesic, and anti-inflammatory effects, likely due to their potent antioxidant and free radical-scavenging activities. A unique isoflavone glycoside, named 4',5,6,7-tetrahydroxy-8-methoxyisoflavone-7-O-beta-D-galactopyranosyl-(1→3)-O-beta-D-xylopyranosyl-(1→4)-O-alpha-L-rhamnopyranoside, has been reported, which shows antimicrobial and antifungal properties.24

Major Compounds Reported:25,26,27

• Luteolin and apigenin (flavonoids) have been detected in methanolic extracts, showing strong free-radical scavenging capacity.

• Chlorogenic acid and caffeic acid have been found as phenolic acids with antioxidant and anti-inflammatory potential.

• β-sitosterol, a plant sterol, was identified and is known for its anti-inflammatory and lipid-lowering effects.

• Squalene, a triterpenoid, has also been reported, which contributes to skin-protective and antioxidant properties.

Phytochemical Screening of Ethanolic Extract of Enhydra Fluctuans

4. Pharmacological Activities

• Antioxidant Activity

The antioxidant potential of the ethanol (EEF), chloroform (CEF), and petroleum ether (PEEF) extracts from the aerial parts of Enhydra fluctuans was estimated through various assays, including reducing power, total phenolic content, DPPH radical scavenging, nitric oxide (NO) scavenging, and superoxide scavenging activities. Among all the tested extracts, the ethanol extract consistently demonstrated the highest antioxidant activity across all evaluations.28 Enhydra fluctuans exhibits significant antioxidant activity, largely attributed to its rich content of phenolic compounds and flavonoids. Various studies revealed that methanolic and ethanolic extracts of its leaves possess strong free radical scavenging abilities in assays such as DPPH, ABTS, and FRAP. These antioxidant effects are likely due to the plant’s ability to donate electrons, neutralize reactive oxygen species (ROS), and prevent oxidative damage to cellular components. The high total phenolic and flavonoid content observed in E. fluctuans extracts further supports its potential as a natural antioxidant source.19,20,27

• Protective Effect

Protective effect of the n-hexane fraction of Enhydra fluctuans aerial against insulitis in alloxan-induced diabetic rats having a body weight of 180-250 g. The n-hexane fraction reduced blood glucose, increased insulin levels, and increased the number of pancreatic β-cells significantly in alloxan-induced diabetic rats. The administration of the n-hexane fraction at a dose of 57.03 mg/kg body weight exerted the best protective effect against insulitis and promoted regeneration of the islet of Langerhans. The results of this study indicate that this herb could effectively reduce insulitis and promote the regeneration of pancreatic tissue in diabetes. The result showed that E. fluctuans has the potential to be developed as a novel diabetes drug.29 Enhydra fluctuans has shown promising antidiabetic potential in various preclinical studies. The plant's extracts, particularly the methanolic and aqueous forms, have demonstrated significant blood glucose-lowering effects in streptozotocin (STZ)-induced diabetic rat models. This hypoglycemic activity is believed to be mediated through mechanisms such as enhancing insulin secretion, improving glucose uptake, and inhibiting carbohydrate-digesting enzymes like α-amylase and α-glucosidase. The presence of bioactive compounds such as flavonoids, alkaloids, and phenolic acids may contribute to these effects, supporting its traditional use in managing diabetes mellitus.30,31,32

• Analgesic and Anti-Inflammatory Activity

Ethanolic extract of Enhydra fluctuans produces analgesic and anti-inflammatory activity. Activity was evaluated by the hot plate method, and anti-inflammatory activity was evaluated by formalin-induced paw edema in Wistar rats. Ethanolic extract at a dose of 300 mg/kg reduces formalin-induced inflammation and also increases pain threshold in rats evidenced by the hot plate method. Experimental results concluded that Enhydra fluctuans has significant analgesic and anti-inflammatory activity due to the presence of flavonoids and phenolic compounds.33 Enhydra fluctuans has demonstrated notable analgesic and anti-inflammatory properties in experimental studies. Ethanolic and aqueous extracts of the plant have shown significant pain-relieving effects in animal models using methods like the hot plate and acetic acid-induced writhing tests. Additionally, anti-inflammatory effects have been observed in carrageenan-induced paw edema models, suggesting inhibition of inflammatory mediators such as prostaglandins and cytokines. These effects are likely due to the presence of flavonoids, alkaloids, and triterpenoids, which possess well-known anti-inflammatory and analgesic actions.34,35,36

• Hepatoprotective Activity

Hepatoprotective activity was studied using the aerial part of Enhydra fluctuans extract against CCl4-induced hepatotoxicity in Wistar albino rats. The extract was found to significantly decrease the CCl4-induced elevation of SGOT, SGPT, bilirubin, and total cholesterol at a dose of 200 mg/kg body weight. But increase HDL-cholesterol and liver weight. concerning the CCl4 toxic rat. The histopathological profile of CCl4-induced hepatotoxic liver revealed extensive centrilobular necrosis.37 Enhydra fluctuans has shown promising hepatoprotective activity in experimental models of liver damage. Studies using carbon tetrachloride (CCl?)- and paracetamol-induced hepatotoxicity in rats have demonstrated that extracts of E. fluctuans significantly reduce elevated liver enzyme levels such as ALT, AST, and ALP, and improve histopathological features of liver tissue. The hepatoprotective effect is attributed to the antioxidant and free radical-scavenging properties of phytoconstituents like flavonoids, phenolics, and saponins, which help protect hepatocytes from oxidative stress and inflammation.38,39,40

• Antimicrobial and Cytotoxic Activity

Three medicinal plants (Enhydra Fluctuans, Clerodendrum viscozum, and A. peniculata) for antimicrobial and cytotoxic activities against some pathogenic organisms and Artemia salina (brine shrimp nauplii). All three showed activity against brine shrimp nauplii. A methanol leaf extract of E. fluctuans showed significant efficacy against the tested bacteria and fungi.41

Enhydra fluctuans has demonstrated noteworthy antimicrobial and cytotoxic properties in various experimental studies. The methanolic and ethanolic extracts of the plant have shown inhibitory effects against a range of bacterial strains such as Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa, as well as fungal pathogens like Candida albicans. These effects are likely due to the presence of bioactive compounds such as flavonoids, alkaloids, and tannins that disrupt microbial cell membranes. Additionally, cytotoxic activity has been observed in brine shrimp lethality assays, indicating the potential for anticancer or antiproliferative applications.42,43,44

• CNS Depressant Activity

Benzene, chloroform, and ethyl acetate extracts of Enhydra fluctuans have demonstrated central nervous system (CNS) depressant effects. Research indicates that these extracts significantly reduce spontaneous locomotor activity and exhibit notable sedative, anticonvulsant, and anti-stress properties in Swiss albino mice.45 Enhydra fluctuans has exhibited central nervous system (CNS) depressant activity in animal studies, suggesting its potential as a sedative or tranquilizing agent. Methanolic and ethanolic extracts of the plant have been evaluated in murine models using standard behavioral tests such as the open field test, hole-board test, and phenobarbitone-induced sleeping time. Results showed a dose-dependent reduction in locomotor activity and increased duration of sleep, indicating CNS depression. These effects are believed to be due to phytochemicals like flavonoids, saponins, and alkaloids, which may act through modulation of GABAergic pathways.46,47

• Anticancer Activity

The flavonoids obtained from Enhydra fluctuans against Ehrlich’s ascites carcinoma-bearing Swiss albino mice showed anticancer activity. Anticancer activity assessed by measuring tumor growth response, percentage increase of life span, haematological parameters, and lipid peroxidation. Two flavonoids, baicalein 7-6-glucoside and baicalein 7-0 diglucoside, treatment with FEF caused a significant decrease in the tumor cell volume and an increase in life span.48 Enhydra fluctuans has shown potential anticancer activity in preliminary studies, primarily through its cytotoxic effects on cancer cell lines and in brine shrimp lethality assays. Extracts of the plant—particularly methanolic and ethanolic leaf extracts—have demonstrated dose-dependent cytotoxicity against cancer cells such as HeLa (cervical cancer) and MCF-7 (breast cancer), suggesting apoptosis-inducing potential. The anticancer activity is attributed to the presence of bioactive compounds like flavonoids, tannins, and saponins, which may interfere with cancer cell proliferation, induce oxidative stress, or trigger programmed cell death.43,44,49

• Anthelmintic Activity

In-vitro anthelmintic activity of ethanolic extract of leaves of Enhydra fluctuans using Phretima posthuma and Tubifex tubifex as test worms. The time of paralysis and time of death were studied, and activity was compared with albendazole as a reference standard. The ethanolic extract of leaves of Enhydra fluctuans exhibited significant anthelmintic activity as evidenced by decreased paralyzing time and death time of the test worm.50 Enhydra fluctuans has demonstrated effective anthelmintic activity in various in vitro studies. Ethanolic and aqueous extracts of the plant have shown dose-dependent paralysis and death of helminths such as Pheretima posthuma (earthworm), which is commonly used as a model for anthelmintic testing. The observed activity is believed to be due to the presence of secondary metabolites like alkaloids, tannins, and flavonoids, which can interfere with the energy metabolism of parasites or damage their integument. These findings support the traditional use of E. fluctuans in treating intestinal worm infections.51,52,53

• Antidiarrhoeal Activity

Methanol and aqueous extract of Enhydra fluctuans showed antidiarrhoeal activity in castor oil-induced diarrhoea. The methanolic extract moderately inhibited the growth of Shigella flexneri, while the aq. Extract inhibited the growth of Staphylococcus aureus and Shigella dysenteriae. Both the extracts at higher dose levels significantly reduced the intestinal transit of charcoal meal in mice. All the results indicated that the extract of Enhydra fluctuans possesses antidiarrhoeal activity.54 Enhydra fluctuans has shown promising antidiarrhoeal activity in experimental models, supporting its traditional use in treating gastrointestinal disorders. Studies using castor oil-induced diarrhoea in rats demonstrated that both ethanolic and methanolic extracts of the plant significantly reduced the frequency of defecation and the weight of fecal matter. The activity is likely due to the presence of flavonoids, tannins, and alkaloids, which may inhibit intestinal motility and reduce gastrointestinal secretions by interfering with prostaglandin pathways. These findings suggest that E. fluctuans possesses compounds with spasmolytic and fluid-absorption enhancing properties. 39,55

• Analgesic Activity

Methanol extract of Enhydra fluctuans whole plant showed analgesic activity on Swiss albino mice by acetic acid induced writhing method and tail flick method using analgesiometer with little modification. The methanolic extract of Enhydra fluctuans given orally at the doses of 250 and 500 mg/kg significantly and dose-dependently reduced the no. of acetic acid-induced writhing in mice.56 Enhydra fluctuans has demonstrated significant analgesic activity in experimental animal models. The ethanolic and methanolic extracts of the plant have been shown to reduce pain responses in the acetic acid-induced writhing test and the formalin-induced pain model. These analgesic effects are likely due to the presence of phytochemicals such as flavonoids and alkaloids, which may act by inhibiting the release of pain-mediating chemicals like prostaglandins or by modulating pain receptors. The results suggest that E. fluctuans may offer potential as a natural alternative for pain management.2034,

5. Nutritional Value

Enhydra fluctuans is a rich source of essential nutrients, including proteins, carbohydrates, and a variety of vitamins and minerals. The leaves of the plant contain notable amounts of vitamin C, vitamin A, and several B-vitamins, which contribute to its antioxidant properties. It is also high in minerals such as calcium, iron, and magnesium, which are crucial for bone health, blood oxygenation, and overall cellular functions. Furthermore, the plant provides dietary fiber, which supports digestive health. The nutritional profile of E. fluctuans makes it a valuable addition to the diet, especially in regions where it is traditionally consumed.53,57  Enhydra fluctuans is not only valued for its medicinal properties but is also widely consumed as a leafy vegetable in many parts of South and Southeast Asia. It is nutritionally rich and serves as a good source of macro- and micronutrients.

• Rich in calcium and iron – supports bone health and helps prevent anemia.

• High in vitamin C and beta-carotene – provides antioxidant protection and supports immune function.

• Low in calories and fat – ideal for weight-conscious or diabetic diets.

• Good source of dietary fiber – aids digestion and helps maintain gut health.

• Contains essential minerals, like potassium and magnesium, beneficial for cardiovascular and muscular function.58

6. Macronutrient and micronutrient profile57,59,60

Macronutrient profile:

• Proteins: The leaves of Enhydra fluctuans are a good source of plant-based proteins, with significant amino acids such as glutamic acid, aspartic acid, and leucine.

• Carbohydrates: The plant contains a moderate amount of carbohydrates, which serve as an energy source. The carbohydrate content is mostly composed of polysaccharides like starch.

• Fats: The fat content is relatively low in the leaves, but it contains essential fatty acids, including linoleic acid, which are beneficial for the body.

• Fiber: The plant is rich in dietary fiber, aiding digestion and improving gut health.

• Water: Enhydra fluctuans contains a significant amount of water, making it useful in traditional remedies for hydration and cooling effects.

Micronutrient profile:

Vitamins:

• Vitamin C: Known for its antioxidant properties, vitamin C content in E. fluctuans helps boost immunity and prevent cellular damage.

• Vitamin A: Present in the form of carotenoids, contributing to vision health and skin care.

• B-Vitamins: Includes folic acid, riboflavin, and niacin, which play essential roles in metabolic processes and energy production.

Minerals:

• Calcium: Vital for bone health, the plant contains a good amount of calcium.

• Iron: Important for oxygen transport and blood health, E. fluctuans provides iron, which is particularly beneficial for preventing iron deficiency anemia.

• Magnesium: Supports muscle and nerve function, and regulates blood sugar levels.

• Phosphorus: Essential for maintaining healthy bones and teeth.

Other Micronutrients: Trace amounts of potassium, zinc, and copper are also present, contributing to various enzymatic functions and maintaining overall health. Approximate Nutritional Composition (per 100 grams of fresh leaves) 61

7. Edible Uses of the Plant

In several regions where Enhydra fluctuans is found, parts of the plant are used in culinary practices.62

i. Traditional dishes sometimes incorporate the tender leaves and young shoots of Enhydra fluctuans L., treating it as a leafy vegetable.63 It is often blanched and added to stir-fries, broths, or stews.

ii. In countries like Malaya, the youthful parts of the plant are eaten raw as salad or lightly cooked before consumption.

iii. In the Philippines, the foliage is crushed and applied externally to alleviate symptoms of cold sores.

iv. The leaves are also considered helpful for treating dermatological issues and certain nervous system conditions.

v. In Bengal, it is commonly washed, chopped, and cooked as a spinach-style dish (sag fry), often boiled with rice and eaten with plain rice, boiled potatoes, and salt.

vi. The plant’s bitter-tasting young parts are known for their mild laxative properties.

vii. The leaves are also pounded into a paste and applied to the scalp to provide a cooling effect.

viii. In Calcutta, the freshly extracted juice of the leaves is sometimes combined with metallic tonic formulations and administered to treat neuralgia and related nerve disorders.

ix. The foliage has antibilious effects, and the juice is used as a soothing agent in cases of gonorrhea, typically mixed with milk from cows or goats.

x. It is also used for feeding cattle, buffalo, goats, sheep, horses, and pigs.

xi. The plant is known to regulate liver function. A decoction is usually prepared the night before, then cooked with rice and served with mustard oil and a pinch of salt.64

8. Toxicological Studies

Toxicological studies on Enhydra fluctuans Lour. have been limited, but existing research provides insights into its safety profile and potential risks.

Acute Toxicity in Zebrafish

An acute toxicity study conducted on zebrafish (Danio rerio) disclosed that the ethanolic leaf extract of Enhydra fluctuans exhibited concentration-dependent lethality. The median lethal concentration (LC??) values at 24, 48, 72, and 96 hours were 204.132, 170.513, 139.478, and 92.956 mg/L, respectively. At the highest concentration of 400 mg/L, 100% mortality was observed within 24 hours. Histopathological examination showed significant damage to the intestine, gills, liver, and brain tissues, including villi rupture, epithelial detachment, and severe vacuolization. Behavioral changes such as reduced swimming, depigmentation, and mucus secretion were also noted.65

Acute Toxicity:

• Studies on the acute toxicity of Enhydra fluctuans have been conducted primarily using oral administration of plant extracts in rodents. In one study, the plant extracts did not show significant signs of acute toxicity at low to moderate doses. However, at higher doses, some animals exhibited mild gastrointestinal distress such as diarrhoea and lethargy.42

• The median lethal dose (LD50) was found to be above 2000 mg/kg body weight for the ethanolic extract in rats, indicating that the plant may not be acutely toxic at typical dosages.

Genotoxicity:

• Research on the genotoxicity of Enhydra fluctuans has been limited, but some studies have suggested that its extracts do not induce chromosomal aberrations or micronuclei formation in mammalian cells.66 Further studies are required to confirm these results and explore the plant’s potential mutagenic effects.

Histopathological Studies:

• Histopathological examinations of vital organs, including the liver, kidneys, and heart, have shown no significant abnormalities after treatment with Enhydra fluctuans extracts at therapeutic doses. However, at higher doses, mild liver congestion and renal tubular dilation were observed, indicating possible organ stress at elevated doses.67

9. Ethnomedicinal Values

Enhydra fluctuans, commonly known as water cress or “Helencha,” holds significant importance in traditional medicine, particularly in South and Southeast Asia. In Ayurveda and folk systems, it is used for managing liver disorders, jaundice, constipation, and skin diseases. Indigenous communities often consume the leaves as a cooling agent and blood purifier. The plant is also traditionally used to treat inflammation, digestive issues, and urinary tract infections. In Bangladesh and parts of India, crushed leaves are applied topically for skin eruptions and wounds, and decoctions are used for treating diarrhea and dysentery. These ethnomedicinal uses are largely attributed to the plant's rich phytochemical content, including flavonoids, alkaloids, and saponins.68,69

The following are the ethnomedicinal values of Enhydra fluctuans:

i. Enhydra fluctuans, a plant widely recognized for its medicinal and nutritional value, is traditionally used across various regions of Northeast India. In Assam’s Cachar district, the stem is employed to treat gastric issues and ulcers, while the entire plant is used as a remedy for constipation.70

ii. Among the Shan tribe descendants of Sino-Tibetan origin from Southwest China, it is considered a wild edible. Studies in Golaghat, Karbi Anglong, Lakhimpur, Dibrugarh, and Jorhat districts reveal its use as a supplementary food, particularly the young shoots consumed as cooked vegetables.71

iii. Locally known as Helechi in Dibrugarh, the plant's aerial parts are crushed and applied to pimples as a traditional treatment.

iv. In Manipur’s Thoubal district, the Meitei-Pangal community refers to the plant as Komprek tujombi and uses it to manage diabetes by boiling the plant (cut at the nodes) and consuming the extract.72

v. Muslim herbalists in the same region use a preparation of boiled leaves mixed with sugar to treat kidney stones.73

vi. In Meghalaya, the Jaintia tribe, who call the plant Kynbat hingcha, utilize the leaf juice for treating skin ailments, as a laxative, and for liver disorders.74,75

vii. Additionally, Enhydra fluctuans is also recognized as a wild edible plant in Assam’s Majuli and Darrang districts.76

viii. Its phagocytic activity has been investigated in studies using samples from Agartala in Tripura state.77

10.Cultivation of Enhydra fluctuans Lour.: Agricultural Practices and Considerations78,79

Enhydra fluctuans Lour., commonly known as buffalo spinach or Helencha, is a semi-aquatic herb belonging to the Asteraceae family. Native to tropical and subtropical regions of Asia, it thrives in humid climates and is cultivated for its edible leaves and medicinal properties.

Climate and Soil Requirements

The plant prefers tropical to subtropical climates with temperatures ranging from 20ºC to 35ºC. it requires high humidity and ample sunlight for optimal growth. Enhydra fluctuans flourishes in most, loamy soils with a slightly acidic to neutral pH (5.5-7.0). while it tolerates waterlogging, stagnant water should be avoided to prevent root rot.

Propagation and Planting

Propagation is typically achieved through stem cuttings, which root readily when placed in moist soil or shallow water. These cuttings should be planted during the monsoon season to take advantage of natural rainfall. Spacing between plants should be maintained at 20-25 cm to allow for adequate growth.

Water Management and Fertilization

Consistent moisture is crucial for Enhydra fluctuans. in dry periods, irrigation should be provided every 2-3 days. The plant benefits from the application of organic fertilizers, such as well-rotted compost or farmyard manure, during land preparation. A balanced dose of NPK fertilizer can further enhance vegetative growth.

Weed and Pest Management

While Enhydra fluctuans is generally resistant to pests and diseases, early-stage weeding is essential to reduce competition. Natural growth habits can suppress weeds over time. Occasional issues with aphids or fungal infections may arise in stagnant or overcrowded conditions, necessitating appropriate management practices.

Harvesting and Post-Harvesting Handling

Harvesting can commence approximately 30-40 days after planting, with subsequent harvests every 15-20 days, depending on growth conditions. The plant should be cut 5-10 cm above the ground to allow for regrowth. Post-harvest, the leaves should be rinsed thoroughly to remove contaminants and packaged in moisture-retaining materials for market distribution.

Economic Potential and Sustainability

Enhydra fluctuations offer significant economic potential for small-scale farmers, particularly in wetland areas. Its rapid growth cycle and multiple harvests per season contribute to its profitability. Additionally, the plant's ability to thrive in low-input systems and its nutritional value make it a sustainable crop choice.

Challenges and Future Prospects

Despite its advantages, challenges such as seasonal limitations and the need for standardized agronomic practices exist. Research into disease-resistant or climate-resilient varieties, as well as the potential for urban aquaponics systems, could further enhance the cultivation of Enhydra fluctuans.

11.Formulation and Extraction Techniques

1. Traditional Formulation Techniques

In traditional medicine systems (such as Ayurveda and folk medicine), Enhydra fluctuans is prepared using simple, non-laboratory techniques:

• Infusion/Decoction

Fresh or dried aerial parts (leaves and stems) are boiled in water to extract bioactive compounds. This preparation is consumed orally for conditions like liver disorders, inflammation, or as a cooling agent.

• Plant Paste

Leaves are ground into a fine paste and applied topically to treat skin ailments such as pimples, rashes, or wounds.

• Juice (Swarasa)

Fresh leaves are crushed and filtered to obtain the juice, which is taken internally to manage digestive issues, high blood pressure, or urinary problems.

• Herbal Diet

The plant is cooked and consumed as a green leafy vegetable, often in combination with other herbs to enhance its medicinal effects.80

2. Laboratory-Based Extraction Techniques:

For phytochemical and pharmacological studies, several solvent extraction methods are commonly used.

Solvent Extraction:

Different solvents are employed based on polarity to extract a broad spectrum of phytochemicals:

• Petroleum Ether Extract (PEEF): Extracts non-polar compounds like fats, oils, and waxes.

• Chloroform Extract (CEF): Targets moderately polar compounds.

• Ethanol Extract (EEF): Effective for polar compounds like phenolics and flavonoids, often shows the highest antioxidant activity.

Procedure

• Drying and Powdering: Aerial parts are cleaned, shade-dried, and ground into coarse powder.

• Soxhlet Extraction: Powdered plant material is subjected to continuous extraction using solvents like ethanol, chloroform, or petroleum ether.

• Filtration and Concentration: Extracts are filtered and concentrated under reduced pressure using a rotary evaporator.

• Storage: The final extract is stored in a cool, dry place for further phytochemical and biological analysis.

Aqueous Extraction:

For a more natural and traditional approach, water is used to extract compounds useful for testing water-soluble constituents and closely mimics traditional formulations.81

Methods of Extraction Used in Studies

In scientific studies, the extraction methods for Enhydra fluctuans Lour. have been optimized to isolate bioactive compounds for various pharmacological evaluations. Below is an overview of the extraction techniques employed in research

1. Soxhlet Extraction Method

A common technique involves the Soxhlet apparatus, which allows for the continuous extraction of compounds from plant material using solvents

Procedure

1. The aerial parts of Enhydra fluctuans are air-dried and powdered.

2. Approximately 1.8 kg of the powdered plant material is extracted successively with petroleum ether (60–80°C) and methanol using the Soxhlet apparatus.

3. The solvents are then removed under reduced pressure to obtain sticky residues.

4. The crude methanolic extract is further partitioned using 10% sulfuric acid solution and successively extracted with chloroform, ethyl acetate, and n-butanol to yield respective fractions.

Outcome

1. This method results in the isolation of various fractions, each containing different bioactive compounds.

2. The ethyl acetate fraction, in particular, has shown the highest antioxidant activity among the fractions tested.82

2. Cold Maceration with Ethanol

For certain studies, a simpler extraction method is employed

Procedure

• The coarse leaf powder is extracted at room temperature with ethanol.

• Solvent-solvent partitioning is then performed to obtain various soluble fractions.

Outcome

• This approach is useful for obtaining extracts rich in flavonoids and other phenolic compounds.

• The ethanolic extract has demonstrated significant antioxidant activity, comparable to standard antioxidants like ascorbic acid.83,84

3. Thin Layer Chromatography (TLC) for Flavonoid Isolation

To isolate specific compounds such as flavonoids

 Procedure

• Ethanolic extracts are subjected to thin layer chromatography using a suitable solvent system.

• This method helps in the separation and identification of flavonoid compounds present in the extract.

Outcome

• The presence of flavonoids is confirmed, which are believed to contribute to the plant's anti-inflammatory and antioxidant properties. These extraction methods are crucial in isolating and characterizing the bioactive compounds of Enhydra fluctuans, facilitating the assessment of its pharmacological potentials.85

12. Formulations in Commercial Products

While Enhydra fluctuans is widely used in traditional medicine, its incorporation into commercial pharmaceutical products is limited. However, the growing interest in herbal supplements has led to the development of various dosage forms.

1. Capsules and Tablets:

Extracts of Enhydra fluctuans are encapsulated or compressed into tablets for standardized dosing. These formulations are designed to provide consistent therapeutic effects and are accessible for consumers.

2. Herbal Teas:

Dried leaves or powdered extracts are used to prepare herbal teas. These are consumed for their potential health benefits, including antioxidant and anti-inflammatory properties.

3. Topical Preparations:

Creams, ointments, or gels containing Enhydra fluctuans extracts are applied to the skin for conditions like acne, eczema, or other inflammatory skin disorders.

4. Liquid Extracts and Syrups:

Concentrated liquid extracts are formulated into syrups or tinctures, offering an alternative for those who prefer liquid dosages.86,87

CONCLUSION

Enhydra fluctuans Lour., a semi-aquatic leafy herb with longstanding use in traditional medicine, is gaining attention in the scientific community due to its rich array of phytochemicals and potential health benefits. This plant is known to contain various bioactive constituents such as flavonoids, alkaloids, saponins, and phenolic compounds that contribute to its diverse pharmacological properties, including antioxidant, anti-inflammatory, antimicrobial, liver-protective, and neuroprotective effects. Traditionally, it has been employed in the management of liver disorders, skin diseases, digestive problems, and neurological conditions, especially in folk medicinal systems of South and Southeast Asia. Beyond its therapeutic applications, E. fluctuans also holds nutritional value, offering essential vitamins and minerals, which supports its categorization as a functional food. Despite its adaptability to moist environments and widespread traditional use, there is a lack of standardized cultivation techniques and agronomic studies to enhance its medicinal yield and quality. Preliminary toxicology reports indicate it is safe for consumption; however, comprehensive safety evaluations over extended periods are still necessary.

While current studies emphasize the potential of E. fluctuans, several critical knowledge gaps remain. There is an urgent need for systematic investigations, including clinical trials, advanced phytochemical analyses, and the formulation of standardized plant extracts to validate traditional knowledge and facilitate its application in modern healthcare.

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