Green Fabrication of Silver Nanoparticles Using Casytha filiformis Extract a Promising Approach for Antimicrobial and Antioxidant Activities

The increasing prevalence of antibiotic-resistant pathogens and oxidative stress-related diseases has posed a significant challenge to public health, prompting the exploration of alternative therapeutic strategies[1]. Nanotechnology, particularly the synthesis of silver nanoparticles (AgNPs), has emerged as a promising solution due to their unique physicochemical properties and potent biological activities, including broad-spectrum antimicrobial and antioxidant effects [2]. Conventional methods of AgNP synthesis typically involve chemical reducing agents and high energy inputs, which can lead to environmental toxicity and biohazard risks [3]. In contrast, green synthesis approaches utilizing plant extracts offer a sustainable, low-cost, and environmentally benign alternative. These methods exploit the phytochemical constituents of plants—such as flavonoids, terpenoids, phenolic acids, and alkaloids—which serve as natural reducing and capping agents in nanoparticle formation [4]

Cassytha filiformis, a parasitic plant known for its traditional medicinal applications, has been reported to contain a rich array of bioactive compounds including phenolics and flavonoids[5]). Despite its pharmacological potential, the application of C. filiformis in nanotechnology remains underexplored. Leveraging its phytochemical profile could provide a novel route for the green synthesis of AgNPs with enhanced biological efficacy.This study aims to fabricate silver nanoparticles using C. filiformis extract via a green synthesis method and evaluate their antimicrobial and antioxidant properties. The findings could contribute to the development of eco-friendly nanomaterials for biomedical and pharmaceutical applications[5]. Cassytha filiformis is a leafless, twining, climbing, parasitic plant with long, threadlike stems. The stems are yellow-green, brown, or orange, up to 8 m long, and adhere to trees or shrubs by small, round suckers. Its spirally arranged leaves are reduced to tiny scales.A sprawling parasitic vine Cassytha filiformis, is widely distributed throughout the regions of Trophics much along the seashores up to the extent of 300m. It is found to be parasiting on many Other host plants like Acacia, Azadirachta, and Mangifera. [6] The “woevine” is usually found as a cosmopolitan in the region of tropics but sometimes becomes apest of economic importance because of its attachment to the valuable orchard trees and other Ornamental plants by means of its tough threadlike extensive branchlets. This plant is considered To be unique in the family of Lauraceae as it is a parasite. Because of the nature of its particular Characteristics, it is taxonomically classified in a separate tribe.[7] Cassytheae comes within the family Lauraceae and it is represented by the single genus CassythaWhich further describes18 different species under it. The genus derived its name, Cassytha, from The Greek name of Cuscuta. The vine has several common names in the regions of the tropics. For example, South Sea Islanders called this vine as “tentanini” which has the meaning “to go Round and round,” and this seems to be a true descriptive adjective for the plants entwining habit.[7]

BIOLOGICAL PROFILE

Fig. No.2 Cassytha filiformis

Taxonomy:                               

• Domain: Eukaryota.                        
• Kingdom :Plantae
• Subkingdom :Tracheobionta
• Phylum: Spermatophyta
• Subphylum: Angiospermae
• Superdivision :Spermatophyta
• Division :Magnoliophyta
• Class :Magnoliopsida
• Subclass :Magnoliidae
• Order :Laurales
• Family :Lauraceae
• Genus :Cassytha [8]

Synonyms:

• English: Love vine, greek kasytas
• Spanish: Alambrillo, bejuco dorado, bejuco fideo, fideos, tente en el air
• Chinese (Taiwan): Kume, wu-kentaso
• Japanese: Sunazuru
• French: Liane parasyte, liane d‘amité, liane ficelle, liane sans fin, mouttaré,
• Fausse cuscute, cord a violon, vermicelli
• German: Schlingfaden[7]

Biological source:

Cassytha filiformis is an orangish, wiry, parasitic vine in the family Lauraceae.[9]

Geographical Source: It  is found in coastal forests of warm tropical regions worldwide including the India, Americas, Indomalaya, Australasia,  and tropical Africa.[10]

Pathogenicity of C. Filiformis to plants

The science of plant pathology treats plant-parasitic seed Plants as a distinct group of plant pathogens that are able To infect other plants and cause disease. Although many Insects have a parasitic relationship with host plants, the Relationship is traditionally defined as food relationship That does not develop further into a disease relationship. In a disease relationship between a parasite and host.[11]The relations are more intimate; a process of infection Ensues and the physiology and  metabolism of the host Is adversely affected. In this case (C. Filiformis), disease Is caused by the effect of infection of host plants by Specialized attachment and penetrating feeding structures known as haustoria that are found on stems of C. Filiformis and similar parasites.[[12]

Morphology     

Stem

Stems are green to orange color, filiform and its glabrous.

Fig.No .3 Cassytha filiformis Stem

Flowers

Flowers are bisexual flowers,small,sessile and spicat.[7]

Fig.No.4 Cassytha filiformis flower

Leaves

Leaves Modified to Minute scale[7]

Fig.No.4 Cassytha filiformis Leaves

FRUIT

The fruit is smooth and fleshy, spherical and seven mm diameter with single seed.[7]

Fig.No.6  Cassytha filiformis Fruit

CHEMICAL  CONSTUENTS

Fig No 8 Chemical constituents.

Development and spreading

Once the host contact is established, the base will dry Up and the plant will lose the connection with the ground And will be further dependent on the host plant for water, Food and nutrition. The haustoria will penetrate the host Plant part (either stem or leave) and start absorbing water And nutrients. The host plant will eventually be suppressed And sometimes killed. With time, the haustoria will flowerAnd seed – the seed being spread by the wind, by water or By birds [7,9,10]

Worldwide distribution

The worldwide distribution of the plant is shown in Fig As per this map, the plant is available on the Majority of the continents, barring Europe and Antarctica. [7]

PHARMACOLOGICAL ACTIVITY:

Antioxidant activity:

The antioxidant activity of Cassytha filiformis extracts such as hexane, ethyl acetate and Methanol were used for the assessment based on their radical scavenging activity (RSA) using 8 the DPPH assay.The methanolic extract was found to show potent antioxidant activity on Comparision with the standard Butylated hydroytoluene(BHT).  methanolic extracts were Further evaluated by the other methods such as Ferric thiocyanate(FTC) method, Thiobarbituric Acid(TBA) test and Superoxide anion radical scavenging assay. The results obtained from the Above experiment suggested that the methanolic extract of Cassytha filiformis have provided a Promising therapeutic potential and could be further applied as a potential source for the drug Development by the pharmaceutical industries.[13][15]

Diuretic activity:

Aqueous and alcoholic extract of Cassytha filiformis was tested for its diuretic activity in Wister Rats. Total urine output volume and the concentration of Na+, K+ and Cl− ions excretion in the Urine were finally estimated. Aqueous and alcoholic extract of Cassytha filiformis was found to Exhibit a significant diuretic activity by causing the marked increase in the Na+ and K+ Excretion.[8][9]

Anti-trypanasomal activity:

Trypanosomiasis is a potent fatal disease affecting both the human and the other domestic Animals in the regions of tropical Africa and South America . Approximately, it was found that Around 50 million people in 36 African countries were at the risk of getting infection and there Were about 300,000 to 500,000, people currently infected and there were 100 deaths every year Due to the above diseases [10]. .In vitro effect of the crude alkaloid extract and the isolated Compounds such as Actinodaphnine ,Cassythine ,Dicentrine on the infecting organism Trypanosoma brucei brucei was tested and was found that the compounds showed the maximum Inhibitory effect against the organism [[12][9]

Anti-cancer Activity

In cytotoxicity was evaluated for the crude Extract and the isolated compounds: neolistine, dicentrine, cassythine, actinodaphine and camptothecin, Using colorimetric assays, i.e. tetrazolium salt MMT and WST-1, with HL-60, mouse 3T3 fibroblasts, human HeLa And the melanoma Mel5 cell line. The crude extract and actinodaphine, casythine, dicentrine and neolistine compounds were found to display cytotoxic properties on Hela, Mel-5 and HL 60 cancer cells and 3T3-non-cancer cell lines.[7]

Anti-hypertension  Activity

The effects of ethanol extracts of Cassytha filiformis on Hypertension-induced rats were evaluated in Two types Of hypertension: endocrine hypertension and oxidative-stress Hypertension were the subjects of the experiment. Accordingly, endocrine hypertension was induced by a prednisone-salt combination, while oxidative stress hypertension Was induced by prednisone-salt combination and L-Nitro Arginine Methyl Ester. In the work, a Cassytha filiformis Ethanol extract of 5 mg/kg dose was discovered to be have An antihypertension effect according to SBP, DBP and MAP (Systolic blood pressure diastolic blood pressure and mean Arterial pressure) results.[10]

Anti-pyretic, anti-inflammatory activities

In [10], analgesic, anti-pyretic and anti-inflammatory Activities of chloroform and methanol extracts were assessed Using rats. For the analgesic activity, the tail immersion method and Haffner’s tail clip method were applied, and Dichlofenac sodium and extract were found to be equally Effective. As to anti-Pyretic Activity, extract administration Brought about increased reaction in elevated temperature in Paracetamol studies. Regarding anti-inflammatory activity, Paw edema was induced by carrageenan, and diclofenac Sodium and extract administration were seen to decrease The inflammation and odema [16]

Anti-microbial study

An anti-microbial study was undertaken in [1,7] for Methanol, ethyl acetate and n-hexane extracts, using 24 hour Broth cultures of Escherichia coli, Staphylococcus aureusAnd Salmonella spp. The outcome of the work was that The methanol extract displayed anti-microbial effects on Staphylococcus aureus and Salmonella spp.  Moreover, Cassytha filiformis Linn has anti-microbial activity against Candida albicans, Staph. Aureus, E. Coli, and Ps. Aeruginosa .[7]

Anti-malarial

The Anti-malarial study of [20] involved an ethanolic Extract of Cassytha filiformis, and used mice that were Infected by plasmodium berghei. The parasite load was Measured in this study through RBC, WBC counts, PCV And Hb concentration. Herein, Cassytha filiformis administration (especially higher dose 400 mg/kg) in the standard Form p-Alaxine recovered the anemic condition caused by The malarial infection, by improving the RBC, WBC counts, Hb and PCV concentration.[7]

Anti-diabetic activity

An anti-diabetic study using an 80% methanolic extract of Cassytha filiformis on albino mice was done by the researchers of [13]. This induced diabetes with alloxan monohydrate Against Glibenclamide as standard. The Cassytha filiformis Extract dose at 600 mg/kg was shown to reduce the fasting Blood sugar levels in mice. In addition, it has a tolerance Range of 2000 mg/Kg orally.[13] 

Data and Sources of Data

The fresh leaves of cassytha filifomis (Akash beal) were collected from the road side farm , Brahmpuri,Dist chandrapur.

Fig 9.  Cassytha filiformis plant

RESEARCH METHODOLOGY

Method of Extraction

Boiling method

Around 25 g new leaves were dried at room temperature for 48hrs and squashed in fine powder. The squashed leaves were boiled with 200 ml distilled water for 30 Min at that point separated with Whatman No.l filter paper (25 (m). Arranged plant extract wasput away 4 C for further used.

Fig.10  cassytha filiformis powder.

Fig.11 Cassytha filiformis Extract

Method preparation

1. Preparation of 2M.AgN03.

     The 2M Solution of of was Prepared by dissolving 2gm AgN03 Powder in 100ml distilied water and prepared solution was stored in cool and dry place.

2. Biosynthesis of silver nanoparticles using plant extract.

The Aqueous solution of silver nitrate (2MM) was Mixed with leaves extract in 9:1raatio and stir at room temperature and heat it 60-90c. The colour of solution Make change into pale yellow centrifuge the solution in 1500??? for 30 Min.

Fig.12 Solution of cassytha filiformis

Fig.13 Centrifuge machine

Fig 14.Centrifuge solution

Fig.15 Filtration of solution

Fllter the solution in whatsmans filter paper wash the filtered with Mannito intravenous infusion 20% after some time pack the silver nanoperticles in ziplock pouch for evaluation test.\

Ingredients table

Fig.16 Silver nanoparticle

3. Chareterization of silver nanoparticles 

The nanoparticles were characterized by following techniques to learn the size, shape and concentration of AgNPs in different Sample.

UV–Visible spectrophotometer

This investigation was encouraged from the Department of Genetics and Plant Breeding, CCSU, Meerut. The AgNPs were described by UV–Vis twofold shaft spectrophotometer (Lasany LI-295). All spectra were recorded at room temperature, in a quartz cell with 1 cm optical way, to know the dynamic conduct of nanoparticles. The checking range for the examples was 200–800 nm. The spectrophotometer was furnished with ‘‘UV programming” to record and break down the information. The bench-mark rectification of the spectrophotometer was completed by utilizing a clear reference. The examples of AgNPs were examined at 0, 4, 12, 24, 48, 72, 96, and 120 hrs.

RESULTS AND DISCUSSION

UV–V is examination of AgNPs

In this exploration study, firstly AgNPs were described by UV–Vis two fold pillar spectrophotometer (Lasany LI-295). All spectra were recorded at room temperature, in a quartz cell with 1 cm optical way, to know the motor conduct of nanoparticles. Thexamining range for the examples was 200–800 nm. The spectro photometer was outfitted with ‘‘UV prov programming” to record and dissect the information. The standard revision of the spectrophotometer was completed by utilizing a clear reference. The samples of AgNPs was dissected at 0, 4, 12, 24, 48, 72, 96,and 120 hrs. The optical peaks for AgNPs were seen between the scopes of 400 to 480 nm. All the replications of treatment tests demonstrated at 96 hr were given in UV–Vis spectroscopy is commonly used to quantitatively describe natural and inorganic nano sized particles. The sample is illuminated with electromagnetic waves in the bright and obvious extents and the consumed light is examined through the subsequent sprectrum 21,23,24] Nanoparticles have optical properties that are added to appraise, shape, fixation, agglomeration state, and refractive record close the nanoparticle surface thusly, UV–Vis spectroscopy a huge gadget for perceiving and portraying these nanoparticles [3,25,26,27] The peaks extend for silver nanoparticles may shift. In different examinations, the tremendous contrast in peak spectra corresponding to AgNPs have been observed, however by and large, it ranges from 400 nm to 480 nm [28,29,30].