Anthelmintic Efficacy of Luffa cylindrica in Relation to its: Phytoconstituents: Spectroscopic Characterization

One of the most frequent parasite diseases that afflict both humans and animals is helminth infections, also referred to as helminthiasis. Parasitic worms, specifically nematodes (Ascaris lumbricoides, Ancylostoma duodenale, and Trichuris trichiura), cestodes (Taenia spp.), and trematodes (Fasciola hepatica), are the source of these diseases. Over 1.5 billion people worldwide are thought to be afflicted by soil-transmitted helminths, with tropical and subtropical areas with poor sanitation and hygiene having the highest frequency [1].  By lowering weight gain, fertility, and production, helminth infections not only endanger human health but also result in large financial losses for the cattle sector [2]. The clinical manifestations of helminthiasis vary depending on the parasite species but often include abdominal pain, diarrhoea, anaemia, malnutrition, impaired physical development, and cognitive decline in children. Chronic infections can also lead to severe complications, thereby posing a considerable public health burden [3]. Currently, helminth infections are managed using a range of synthetic anthelmintics, including ivermectin, piperazine, mebendazole, and albendazole. These medications mostly work by interfering with the parasites' energy metabolism, preventing neuromuscular activity, or affecting their ability to reproduce [4].  However, helminth resistance has emerged as a result of the ongoing and careless use of these medications, and this is currently seen as a major worldwide problem [5]. Furthermore, synthetic anthelmintics can be expensive, difficult to obtain in remote areas, and occasionally linked to negative side effects [6]. These limitations have prompted a renewed interest in exploring plant-derived anthelmintics as safer, more affordable, and sustainable alternatives. Traditional medicine systems worldwide have documented the use of numerous plants with vermifugal and vermicidal properties. Phytochemicals such as tannins, flavonoids, alkaloids, saponins, and terpenoids have been identified as key contributors to anthelmintic effects, acting through mechanisms such as disruption of parasite membranes, interference with energy metabolism, and paralysis of neuromuscular activity [7]. Because of its ethnomedical use as a vermifuge, Luffa cylindrica (Family: Cucurbitaceae) is particularly interesting in this context. According to earlier phytochemical research, L. cylindrica contains secondary metabolites linked to anthelmintic action [8,9]. Despite its long history of use, there is little scientific evidence to support its effectiveness. Therefore, a thorough assessment of its anthelmintic potential is necessary to support its use as a natural substitute for synthetic medications and to open the door for the creation of innovative plant-based medicines. Modern pharmacological research continues to heavily rely on the centuries-old usage of therapeutic plants in traditional healthcare systems. Among these is the tropical climbing vine Luffa cylindrica (Family: Cucurbitaceae), sometimes referred to as sponge gourd, Egyptian cucumber, or loofah. It is widely grown throughout Asia, Africa, and portions of the Americas. Although its fibrous fruit is well known for its natural scrubber properties, Luffa cylindrica has attracted scientific interest because of its rich phytochemical composition and variety of therapeutic uses [10,11]. Traditionally, various parts of the plant-including the fruit, seeds, roots, and leaves—have been used in the treatment of conditions such as asthma, sinusitis, jaundice, leprosy, haemorrhoids, and intestinal parasites [12,13]. These ethnomedicinal uses are supported by modern phytochemical studies revealing the presence of bioactive constituents like flavonoids, alkaloids, saponins, phenolic acids, terpenoids, and cucurbitacin’s [14-16]. The pharmacological potential of the plant, which includes antibacterial, anticancer, antioxidant, hepatoprotective, anti-inflammatory, and antidiabetic properties, has been the subject of recent studies [17-19]. The way that particular phytoconstituents of Luffa cylindrica interact with cellular targets to produce therapeutic effects has been further clarified by mechanistic investigations, such as molecular docking and in vitro experiments [20]. Given the increasing global interest in plant-based drug discovery and natural product chemistry, Luffa cylindrica presents a promising candidate for pharmaceutical exploration. This review aims to consolidate current findings on the phytochemical constituents of Luffa cylindrica, discuss their mechanisms of pharmacological action, and highlight potential directions for future research in drug development and molecular medicine.

Taxonomic Classification:

Table 1: Taxonomical Classification of luffa cylindrica [21]:

Morphological Characteristics [22,23]:

Habit:  Herbaceous, annual climbing or trailing vine with angular, ridged stems.

Stem:   Pentangular, green, hairy when young, becoming globous with age; climbing with coiled tendrils.

Leaves:  Alternate, palmately lobed (5–7 lobes), rough-textured, 10–20 cm           across.

Tendrils: Branched or unbranched, spring-like, used for climbing support.

Flowers:  Monoecious (male and female flowers on the same plant); yellow, solitary or racemose. Male flowers borne in clusters; female flowers solitary.

Fruit: Cylindrical, elongated (20–60 cm), green with smooth skin, fibrous at maturity; spongy network of vascular bundles forms inside.

Seeds: Flat, smooth, black or dark brown, elliptical in shape; encased in a fibrous network.

Root: Taproot system with lateral branches.

Luffa cylindrica, commonly known as sponge gourd or ribbed gourd, is a fast-growing, perennial, climbing herbaceous vine. The plant is dioecious, with male and female flowers occurring separately. Its stems are green, angular, and hairy, with tendrils arising from the leaf axils that help in climbing. The leaves are large, alternate, simple, and palmately lobed, usually with 3–5 lobes and serrated margins; their surface is rough and hairy. The plant produces unisexual yellow flowers, with male flowers smaller and clustered on long peduncles, while female flowers are larger and usually solitary, giving rise to elongated cylindrical fruits. The fruits are green when immature, turning yellowish-brown upon maturity, with longitudinal ridges and a spongy interior, typically measuring 30–60 cm in length. The seeds are flat, oblong, brown, and hard, arranged along the central fibrous axis of the fruit. Luffa cylindrica has a deep taproot system with fibrous lateral roots, aiding in nutrient and water absorption. Its morphological adaptations, including tendrils and hairy surfaces, facilitate climbing, reduce water loss, and provide some protection against herbivores.

RESULT AND DISCUSSION:

PREPARATION OF PLANT MATERIAL:

Making Extracts: Luffa cylindrica leaves were crushed in an electric blender after being dried in the shade to create coarse powder, and then treated with methanol and water as a solvent for Soxhlet extraction (continuous hot extraction). A rotary evaporator was used to concentrate the extracts, which were then utilized to test for anthelmintic activity.  To determine whether phytoconstituents were present in the extract, a preliminary phytochemical screening was conducted.

Plant material

Luffa cylindrica leaves were handpicked. The leaves were collected in the month of March-May from Luffa cylindrica plant from Kamal Nagar, Bidar district, Karnataka, India. The fresh leaves and flowers were collected for further use. The handpicked leaves were then washed with running tap water for 2-3 times and finally washed with distilled water to remove the dirt and dried under shade for 10- 15 days.

Preparation of Extracts:

The leaves of Luffa cylindrica were dried under shade and crushed in an electric blender to form coarse powder and subjected to Soxhlet extraction (Continuous hot extraction) by using methanol and water as solvent. The extracts were concentrated by rotary evaporator and used for testing anthelmintic activity. Preliminary phytochemical screening was carried out to assess the presence of phytoconstituents in the extract.

Experimental worms: All the experiments were carried out in Indian adult earthworms (Pheretima posthuma) due to its anatomical resemblance with the intestinal roundworm parasites of human beings. They were collected from moist soil and washed with water to remove all fecal matters.

Administration of Albendazole: Albendazole (20 mg/ml) was prepared by using 0.9% w/v of Normal Saline as a Control as administered as per method.

Administration of extract:

The suspension of Methanolic and aqueous extract of leaves of Luffa cylindrica of different concentration (25,50,100,150,200 mg/ml) were prepared by using 0.9% w/v of Normal saline as a control and final volume was made up to 10 ml for respective concentration. Albendazole was used as standard. Five Groups of approximately equal size worms consisting Two of earthworms individually in each group were released into in each 10 ml of desired concentration of drug and extracts in the Petri dish.

Experimental Design:

The anthelmintic activity was performed according to the method. On adult Indian earth worm Pheretima posthuma as it has anatomical and physiological resemblance with the intestinal round worm parasites of human beings. Pheretima posthuma was placed in Petri dish containing different concentrations (25,50,100,150&200 mg/ml) of methanolic extract of leaves of Luffa cylindrica. Each Petri dish was placed with 2 worms and observed for paralysis or death. Mean time for paralysis was noted when no movement of any sort could be observed, except when the worm was shaken vigorously; the time death of worm (min) was recorded after ascertaining that worms neither moved when shaken nor when given external stimuli. The test results were compared with Reference compound Albendazole (20 mg/ml) treated samples.

Figure 1. The Different Concentration of extract With the Earth Worm

Preliminary phytochemical analysis of methanolic extracts showed the presence of Flavonoids, Saponins, Tannins, Steroids, Terpenoids & Alkaloid whereas aqueous revealed the Tannins, Steroids & Alkaloid active phytoconstituents. The data revealed that the methanol extract showed anthelmintic activity at a concentration of 100 mg/ml, showed paralysis and death at similar concentrations. The other test concentrations of the extracts showed marked degree of anthelmintic activity. The anthelmintic effect of extracts is comparable with that of the effect produced by the standard drug albendazole. Parasitic helminths affect animals and man, causing considerable hardship and stunted growth. Hundreds of millions if not billions of human infections by helminths exist worldwide and increased world travel and immigration from the developing countries. However tremendous advances have been made during the previous decade and substantial number of synthetic precursors have been derived to cope up the damage caused by parasite, but unfortunately no effective medicine has been developed so far. Moreover, the problems associated with the use of such drugs like some serious side effects and development of resistance drives the severity of infection to the next level. These factors paved the way for herbal remedies as alternative anthelmintics. Evaluation of activities of medicinal plants claimed for possessing the anthelmintic property is getting the attention these days. Screening and proper evaluation of the claimed medicinal plants could offer possible alternatives that may be both sustainable and environmentally acceptable. The results of this study have shown promising anthelmintic activity suggesting the possible use of L. cylindrica extracts in intestinal nematode control.

Figure 2. Anthelmintic activity of Methanol extract of luffa cylindrica leaves.

FT-IR SPECTROSCOPY:

To identify the major functional groups, present in the methanolic extract and column fractions of Luffa cylindrica based on absorption bands and to determine whether a particular sample of an organic compound is identical to another or not, IR spectroscopy is utilized. This is due to the fact that the infrared spectra of organic compounds show a great number of absorption bands. Thus, two compounds must be samples of the same substance if their IR spectra are identical.