Laboratory Assessment of Molluscicidal Activities of Cannabis sativa,Acacia nilotica,and Tinospora cordifolia Against Snail Host of Fasciola spp.

Abstract

Background:

The potential molluscicidal extracts, obtained from indigenous plants Cannabis sativa, Acacia nilotica, and Tinospora cordifolia, were tested for toxicity against freshwater pulmonate snail Lymnaea acuminata, an intermediate host of Fasciola hepatica. The organic extracts had a significant effect on young snails.

Materials and Methods:

All organic extracts and column-purified fractions gave median lethal concentrations (19–100.05 mg/L; 24 h) that fell well within the threshold level of 100 mg/L, set for a potential molluscicide by the World Health Organization.

Results:

The toxicity of T. cordifolia stem acetone extract (96 h LC₅₀: 16.08 mg/L) was more pronounced compared with C. sativa leaf ethanol extract (96 h LC₅₀: 16.32 mg/L) and A. nilotica leaf ethanol extract (96 h LC₅₀: 24.78 mg/L). β-caryophyllene, gallic acid, and berberine were characterized and identified as active molluscicidal components. Co-migration of β-caryophyllene (retardation factor [Rf] 0.95), gallic acid (Rf 0.30), and berberine (Rf 0.23) with column-purified parts of Cannabis sativa, Acacia nilotica, and Tinospora cordifolia on thin-layer chromatography demonstrates same Rf value, that is, 0.95, 0.30, and 0.23, respectively.

Conclusion:

This study indicates that these extracts thus represent potential plant-derived molluscicides that are worthy of further investigations.

Introduction

Fascioliasis, a parasitic disease of animals as well as human beings, is widespread in tropical and subtropical countries and caused by trematodes Fasciola hepatica and Fasciola gigantica (Singh et al., 2012). Pulmonate snail Lymnaea acuminata serves as intermediate hosts for Fasciola spp. (Mas-Coma et al., 2005). Fascioliasis causes great economic losses to cattle, water buffalo, sheep, and goat farmers across the world (Portugaliza et al., 2019; Singh et al., 2012). The disease affects animal conditions and production is reduced (Mazeri et al., 2017). Human fascioliasis is reported worldwide and characterized by abdominal pain, hypereosinophilia, and exceptionally acute pancreatitis (Echenique-Elizondo et al., 2005; Fürst et al., 2012; Yattoo et al., 2021).

One of the possible solutions to control fasciolosis is to disrupt the life cycle of Fasciola by killing the intermediate host snails below the threshold level (Singh et al., 2012). Out of several methods to eliminate the snail population, one method of choice is the application of commercially available synthetic molluscicides. However, the high cost of synthetic molluscicides and their negative impacts on the environment, including their toxicity to nontarget organisms like fish, as well as the development of snail resistance to these compounds have given a new impetus to the study of eco-friendly and cost-effective molluscicides of plant origin (Adetunji and Salawu, 2010).

A previous study by the authors demonstrated the lethal action of aqueous extracts of indigenous plants Cannabis sativa, Acacia nilotica, and Tinospora cordifolia against the snail Lymnaea acuminata (Singh and Singh, 2023) (Tables 1 –4). This study has the objectives to evaluate the potential molluscicidal activities of organic extracts of these plant parts and identification of active phytoconstituents responsible for toxicity.

Table 1.

Concentration of Different Plant Products and Their Active Components Used for Toxicity Determination Against Lymnaea acuminata

Parts of plant and their extracts	Concentration (mg/L)
Cannabis sativa leaf (aqueous)	150, 200, 250, 300
Chloroform extract	30, 50, 70, 90
Acetone extract	30, 50, 70, 90
Ethanol extract	15, 20, 25 30
Column purified	9, 12, 15, 20
β-caryophyllene	3, 5, 7, 9
Acacia nilotica leaf (aqueous)	200, 300, 400, 500
Chloroform extract	30, 50, 70, 90
Acetone extract	30, 50, 70, 90
Ethanol extract	20, 30, 40, 50
Column purified	10, 15, 20, 25
Gallic acid	5, 7, 9, 15
Tinospora cordifolia stem (aqueous)	450, 550, 650, 750
Chloroform extract	30, 50, 70, 90
Acetone extract	15, 20, 25 30
Ethanol extract	20, 30, 40, 50
Column purified	9, 12, 15, 25
Berberine	7, 9, 12, 15

Table 2.

Toxicity of Cannabis sativa Leaf, Organic Solvent Extracts, Column-Purified Fraction, and β-Caryophyllene (Active Component) Against Snail Lymnaea acuminata at Different Exposure Periods

Exposure period	Tested materials	LC₅₀ (mg/L)	Limits		Slope value	t-ratio	Heterogeneity
Exposure period	Tested materials	LC₅₀ (mg/L)	LCL	UCL	Slope value	t-ratio	Heterogeneity
24 h	C. sativa leaf (aq.)	304.31	271.84	375.92	4.33 ± 0.86	5.03	0.24
	Chloroform extract	97.48	81.31	138.62	2.82 ± 0.56	4.97	0.33
	Acetone extract	99.90	81.29	148.31	2.58 ± 0.55	4.69	0.23
	Ethanol extract	30.43	27.18	37.59	4.33 ± 0.86	5.03	0.24
	Column purified	19.68	17.27	25.10	3.75 ± 0.73	5.08	0.23
	β-caryophyllene	9.42	7.88	13.21	2.74 ± 0.55	4.97	0.26
48 h	C. sativa leaf (aq.)	248.92	226.48	283.91	4.05 ± 0.78	5.18	0.22
	Chloroform extract	74.27	63.90	93.42	2.63 ± 0.50	5.19	0.21
	Acetone extract	83.22	68.24	123.90	2.08 ± 0.49	4.20	0.21
	Ethanol extract	26.89	22.64	28.39	4.05 ± 0.78	5.18	0.22
	Column purified	15.58	13.99	18.08	3.57 ± 0.68	5.22	0.21
	β-caryophyllene	6.87	5.90	5.58	2.54 ± 0.49	5.12	0.27
72 h	C. sativa leaf (aq.)	197.54	172.66	218.64	3.73 ± 0.76	4.91	0.25
	Chloroform extract	48.60	39.40	57.19	2.33 ± 0.47	4.89	0.52
	Acetone extract	48.89	37.87	59.14	1.99 ± 0.47	4.24	0.21
	Ethanol extract	19.75	17.26	21.86	3.73 ± 0.76	4.91	0.25
	Column purified	12.01	10.36	13.45	3.36 ± 0.67	4.96	0.22
	β-caryophyllene	4.74	3.81	5.58	2.31 ± 0.47	4.86	0.27
96 h	C. sativa leaf (aq.)	160.96	135.00	178.48	4.51 ± 0.82	5.48	0.43
	Chloroform extract	34.60	26.52	40.61	2.90 ± 0.51	5.66	0.52
	Acetone extract	34.55	25.83	40.92	2.72 ± 0.50	5.39	0.41
	Ethanol extract	16.32	13.88	18.01	4.66 ± 0.82	5.65	0.43
	Column purified	9.80	8.29	10.88	4.38 ± 0.76	5.74	0.35
	β-caryophyllene	3.47	2.64	4.08	2.83 ± 0.51	5.56	0.49

Mortality was determined every 24 h up to 96 h. Each set of experiment was replicated six times.

Significant negative regression (p < 0.05) was observed between exposure time and LC₅₀ of treatments.

aq., aqueous; LCL, lower confidence limit; UCL, upper confidence limit.

Table 3.

Toxicity of Acacia nilotica Leaf, Organic Solvent Extracts, Column-Purified Fraction, and Gallic Acid (Active Component) Against Snail Lymnaea acuminata at Different Exposure Periods

Exposure period	Tested materials	LC₅₀ (mg/L)	Limits		Slope value	t-ratio	Heterogeneity
Exposure period	Tested materials	LC₅₀ (mg/L)	LCL	UCL	Slope value	t-ratio	Heterogeneity
24 h	A. nilotica leaf (aq.)	535.98	463.48	704.34	3.63 ± 0.70	5.16	0.24
	Chloroform extract	98.70	82.96	137.06	3.04 ± 0.59	5.12	0.25
	Acetone extract	100.05	82.32	155.40	2.46 ± 0.54	4.53	0.19
	Ethanol extract	53.59	46.34	70.43	3.63 ± 0.70	5.16	0.24
	Column purified	27.27	23.30	37.18	3.37 ± 0.68	4.94	0.20
	Gallic acid	15.10	12.52	21.45	2.84 ± 0.54	5.23	0.28
48 h	A. nilotica leaf (aq.)	429.67	378.26	524.62	3.14 ± 0.60	5.17	0.27
	Chloroform extract	74.94	65.14	97.76	2.61 ± 0.50	5.14	0.28
	Acetone extract	73.47	62.43	95.04	2.38 ± 0.49	4.80	0.13
	Ethanol extract	42.96	37.82	52.46	3.14 ± 0.60	5.17	0.24
	Column purified	21.48	18.91	26.23	3.14 ± 0.60	5.17	0.27
	Gallic acid	11.16	9.62	14.02	2.17 ± 0.51	5.37	0.19
72 h	A. nilotica leaf (aq.)	323.46	285.36	364.42	3.33 ± 0.58	5.57	0.53
	Chloroform extract	53.96	65.14	96.76	2.47 ± 0.48	5.62	0.55
	Acetone extract	72.20	38.38	55.17	2.43 ± 0.48	5.06	0.43
	Ethanol extract	32.34	28.52	36.44	3.33 ± 0.58	5.67	0.53
	Column purified	16.02	14.04	18.12	3.20 ± 0.58	5.49	0.58
	Gallic acid	8.01	7.03	9.10	3.14 ± 0.51	6.07	0.28
96 h	A. nilotica leaf (aq.)	247.89	208.51	278.40	3.62 ± 61	5.93	0.67
	Chloroform extract	39.03	31.53	44.99	2.96 ± 0.50	5.87	0.70
	Acetone extract	33.21	24.53	39.48	2.76 ± 0.51	5.39	0.63
	Ethanol extract	24.78	20.85	27.84	3.62 ± 0.61	5.93	0.67
	Column purified	12.39	10.33	13.97	3.48 ± 0.60	5.76	0.58
	Gallic acid	6.10	5.22	6.82	3.72 ± 0.59	6.25	0.36

Mortality was determined every 24 h up to 96 h. Each set of experiment was replicate six times.

Significant negative regression (p < 0.05) was observed between exposure time and LC₅₀ of treatments.

Table 4.

Toxicity of Tinospora cordifolia Stem, Organic Solvent Extracts, Column-Purified Fraction, and Berberine (Active Component) Against Snail Lymnaea acuminata at Different Exposure Periods

Exposure period	Tested materials	LC₅₀ (mg/L)	Limits		Slope value	t-ratio	Heterogeneity
Exposure period	Tested materials	LC₅₀ (mg/L)	LCL	UCL	Slope value	t-ratio	Heterogeneity
24 h	T. cordifolia stem (aq.)	772.62	706.98	919.00	5.81 ± 1.18	4.92	0.19
	Chloroform extract	90.16	77.53	116.92	3.18 ± 0.58	5.45	0.22
	Acetone extract	29.67	26.93	35.01	4.97 ± 0.90	5.50	0.22
	Ethanol extract	49.71	43.79	61.79	3.7 ± 0.69	5.48	0.21
	Column purified	22.98	19.59	30.46	3.06 ± 0.56	5.41	0.34
	Berberine	14.65	13.11	17.72	4.33 ± 0.78	5.55	0.23
48 h	T. cordifolia stem (aq.)	651.25	606.67	719.66	5.39 ± 1.05	5.10	0.19
	Chloroform extract	66.52	57.2	81.22	2.54 ± 0.49	5.16	0.29
	Acetone extract	24.39	22.20	27.62	4.06 ± 0.77	5.22	0.27
	Ethanol extract	38.49	33.98	45.41	3.06 ± 0.58	5.19	0.28
	Column purified	16.71	14.51	20.24	2.70 ± 0.52	5.11	0.33
	Berberine	11.68	10.53	13.41	3.66 ± 0.69	5.29	0.23
72 h	T. cordifolia stem (aq.)	552.65	504.35	593.78	5.30 ± 1.03	5.10	0.19
	Chloroform extract	45.54	36.38	53.41	2.38 ± 0.47	4.98	0.29
	Acetone extract	19.26	16.78	21.26	3.85 ± 0.76	5.04	0.26
	Ethanol extract	28.11	23.37	32.07	2.88 ± 0.57	5.01	0.27
	Column purified	11.86	9.80	13.63	2.74 ± 0.54	5.03	0.25
	Berberine	9.00	7.76	10.03	3.52 ± 0.68	5.10	0.23
96 h	T. cordifolia stem (aq.)	484.06	436.51	517.17	6.81 ± 1.14	5.95	0.55
	Chloroform extract	33.87	26.50	53.41	3.16 ± 0.52	5.97	0.46
	Acetone extract	16.08	13.88	17.63	5.20 ± 0.86	6.05	0.41
	Ethanol extract	22.44	18.05	24.97	3.86 ± 0.64	6.02	0.43
	Column purified	9.49	7.93	10.62	4.19 ± 0.71	5.90	0.32
	Berberine	7.42	6.36	8.18	4.87 ± 0.80	6.09	0.36

Mortality was determined every 24 h up to 96 h. Each set of experiment was replicate six times.

Significant negative regression (p < 0.05) was observed between exposure time and LC₅₀ of treatments.

Materials and Methods

The experiment was carried out in the Malacology laboratory, Department of Zoology, D.D.U. Gorakhpur University, Gorakhpur.

Plants used

Fresh aerial parts of Cannabis sativa (leaf), Acacia nilotica (leaf), and Tinospora cordifolia (stem) (Fig. 1) were collected from local areas and identified by the Department of Botany, DDU Gorakhpur University, Gorakhpur, India. Plant parts were washed thoroughly, shade dried, and pulverized separately in the electric grinder, and the crude powders obtained were then sieved with the help of fine mesh cloth. These fine powders were used separately for toxicity experiments.

FIG. 1.

(A) Cannabis sativa leaf, (B) Acacia nilotica leaf, and (C) Tinospora cordifolia stem.

Solvent extraction

Fifty grams of C. sativa (leaf), A. nilotica (leaf), and T. cordifolia (stem) was extracted separately with 100 mL of each solvent viz. chloroform (99%), acetone (99%), and ethanol (95%) at room temperature for 24 h. Each preparation was filtered through Whatman No.1 filter paper and the filtrates were subsequently evaporated under a vacuum (Jaiswal and Singh, 2008). The residues, thus obtained, were used for the determination of molluscicidal activity.

Column purification

One hundred milliliters of Cannabis sativa leaf ethanol extract, Acacia nilotica leaf ethanol extract, and Tinospora cordifolia stem ethanol extract was subjected to silica gel (60–120 mesh, Qualigens glass, Precious Electro Chemindus Private Limited, Mumbai, India) chromatography through 95 × 45 cm column. Fifty-five fractions of C. sativa were eluted with methanol (95%) and 55 fractions of A. nilotica and Tinospora cordifolia were eluted with ethanol (95%). Solvents were evaporated under a vacuum and the remaining solids thus obtained were used for determination of molluscicidal activity of each fraction.

Thin-layer chromatography

Thin-layer chromatography (TLC) was performed by the method of Singh and Singh (1995) with some modifications to identify the active component present in C. sativa leaf, A. nilotica leaf, and T. cordifolia stem extracts. TLC was done on 20 × 20 cm of precoated silica gel (Precious Electrochemical Industry Pvt. Ltd., Mumbai, India). For C. sativa, the solvent n-hexane/ethyl acetate (3:1 v:v) was used as the mobile phase, and the solvent benzene/ethyl acetate (9:1 v:v) was used as the mobile phase for A. nilotica and T. cordifolia. Spots of column-purified fractions of C. sativa leaf, A. nilotica leaf, and T. cordifolia stem extracts, along with their respective active components β-caryophyllene, gallic acid, and berberine (Fig. 2), were applied on TLC plates with the help of a micropipette. Furthermore, the TLC plates were developed by spraying fast blue reagent for C. sativa and iodine vapor for A. nilotica and T. cordifolia. The plates were immediately traced to create copies of the chromatogram, after which the retardation factor (Rf) was determined.

FIG. 2.

(A) β-caryophyllene (C. sativa), (B) gallic acid (A. nilotica), and (C) berberine (T. cordifolia) (Source. https://www.sigmaaldrich.com/IN/en/search).

Pure compound

β-caryophyllene (1R,4E,9S)-4,11,11-trimethyl-8-methylidenebicyclo[7.2.0]undec-4-ene), gallic acid (3,4,5-Trihydroxybenzoic acid), and berberine (9,10-Dimethoxy-7,8,13,13a-tetradehydro-2′H-[1,3]dioxolo[4′,5′:2,3]berbin-7-ium) were procured from Sigma Aldrich Chemicals Private Limited, Maharashtra, India.

Animal collection

Adult and healthy Lymnaea acuminata (1.35 ± 0.2 cm in length) were collected from freshwater ponds of Gorakhpur District, Uttar Pradesh, India, and were acclimatized for 72 h in the laboratory conditions. Ten snails were then allocated to each experimental group and immersed in 3 L dechlorinated tap water (23 ± 1°C). The pH, free carbon dioxide, dissolved oxygen, and bicarbonate alkalinity were 7.1–7.3, 5.2–6.3, 6.6–7.3, and 102–104 mg/L, respectively. The dead snails were removed from the aquariums to avoid any contamination.

Toxicity experiment

Concentration–response relationship

The toxicity experiments were performed by the method of Singh and Singh (2023). Ten snails were kept in a glass aquarium containing 3 L of dechlorinated tap water and were exposed continuously for 96 h to different concentrations of C. sativa leaf, A. nilotica leaf, and T. cordifolia stem extracts (Table 1). The experiment was set up in six replicates. Negative control was set up under similar conditions without any treatment. Mortality of snails was recorded at intervals of 24 h up to 96 h and death of the snails was determined and confirmed by lack of any movement and lack of reaction to irritation of foot with needle probe to elicit typical withdrawal movements and contraction of their body in the shell.

Statistical analysis (p < 0.05)

The LC₅₀ values, lower confidence limits, upper confidence limits, slope values, and heterogeneity factor, were calculated by polo software program (PoLo Plus LeOra software version 2.0).

Results

The toxicity of different organic solvent extracts of C. sativa leaf, A. nilotica leaf, and T. cordifolia stem was time and concentration dependent. The number of deaths among L. acuminata snails treated with organic solvent extracts, column-purified fractions, and pure compounds increased with increased treatment concentrations (Figs. 3 –5). The calculated values of LC₅₀ of organic extracts and column-purified fractions of all three plant species against adult Lymnaea acuminata are given in Tables 2–4. T. cordifolia stem acetone extract was found to be most promising after 96 h of exposure (LC₅₀ 16.08 mg/L). The values indicate that the C. sativa leaf ethanol extract (24 h LC₅₀: 30.43 mg/L), A. nilotica leaf ethanol extract (24 h LC₅₀: 53.59 mg/L), and T. cordifolia stem acetone extract (24 h LC₅₀: 29.67 mg/L) were more toxic in comparison to other organic extracts (Tables 2–4). The column-purified fractions of these plants were highly toxic. The 96-h LC₅₀ of the column-purified fraction of T. cordifolia (9.49 mg/L) was higher compared with C. sativa (9.80 mg/L) and A. nilotica (12.39 mg/L) (Tables 2–4). The 96-h LC₅₀ of β-caryophyllene, gallic acid, and berberine was 3.47, 6.10, and 7.42 mg/L, respectively. TLC analysis demonstrated that the Rf values of β-caryophyllene (0.95) were equivalent to the Rf value of the column-purified fraction of C. sativa (0.95), gallic acid (0.30) was equivalent to the Rf value of the column-purified fraction of A. nilotica (0.30), and berberine (0.23) was equivalent to the Rf value of the column-purified fraction of T. cordifolia (0.23). Mortality rates in snail L. acuminata were over 40% for each treatment and highest mortality rate recorded for T. cordifolia leaf acetone extract (98.3%) (Table 5).

FIG. 3.

Average mortality of snail Lymnaea acuminata on exposure of different solvent extracts, column-purified fraction of C. sativa leaf and active component β-caryophyllene.

FIG. 4.

Average mortality of snail L. acuminata on exposure of different solvent extracts, column-purified fraction of A. nilotica leaf and active component gallic acid.

FIG. 5.

Average mortality of snail L. acuminata on exposure of different solvent extracts, column-purified fraction of T. cordifolia stem, and active component berberine.

Table 5.

Molluscicidal Activity of Indigenous Plant Extracts and Their Active Components on Lymnaea acuminata After 96 h Exposure

Treatment	%Mortality
Cannabis sativa (Leaf)
Chloroform extract	48.0 (30)	60.0 (50)	76.0 (70)	95.0 (90)
Acetone extract	48.0 (30)	60.0 (50)	75.0 (70)	93.3 (90)
Ethanol extract	48.3 (15)	60.0 (20)	75.0 (25)	95.0 (30)
Column purified	48.3 (9)	60.0 (12)	75.0 (15)	95.0 (20)
β-caryophyllene	48.3 (3)	60.0 (5)	75.0 (7)	95.0 (9)
Acacia nilotica (Leaf)
Chloroform extract	43.3 (30)	55.0 (50)	66.6 (70)	96.6 (90)
Acetone extract	51.6 (30)	60.0 (50)	75.0 (70)	96.6 (90)
Ethanol extract	43.3 (20)	55.0 (30)	66.6 (40)	96.6 (50)
Column purified	43.3 (10)	55.0 (15)	66.6 (20)	95.0 (25)
Gallic acid	43.3 (5)	55.0 (7)	66.6 (9)	96.6 (15)
Tinospora cordifolia (Stem)
Chloroform extract	48.3 (30)	63.3 (50)	80.0 (70)	96.6 (90)
Acetone extract	48.3 (15)	63.3 (20)	80.0 (25)	98.3 (30)
Ethanol extract	48.3 (20)	63.3 (30)	80.0 (40)	96.6 (50)
Column purified	48.3 (9)	63.3 (12)	80.0 (15)	96.6 (25)
Berberine	48.3 (7)	63.3 (9)	80.0 (12)	96.6 (15)

All data in the table are mean of six replicates (n = 10). Values in parentheses are treatment dose in mg/L.

The slope values given in Tables 2–4 were steep. Based on each of the six replicates, the separate estimates of LC values were found to be within the 95% confidence limit of LC₅₀. The heterogeneity factor was less than 1.0, the t-ratio was higher than 1.96, and the g-value was <0.5 at all probability levels (90, 95, and 99). There was a significant negative regression (p < 0.05) between exposure time and the LC₅₀ of exposures (Table 2–4).

Discussion

Sustainable control of fascioliasis requires that the life cycle of Fasciola hepatica should be interrupted by killing the intermediate host snails. Niclosamide, trifenmorph, bayluscide, and the other synthetic molluscicides currently in use to control the aquatic host snails and larvae of Fasciola spp. are toxic to nontarget organisms like fish and very expensive, and resistance to snails is highly probable (Marston and Hostettmann, 1985). Plant molluscicides may serve as cheap and biodegradable, effective alternatives to synthetic molluscicides and may be obtained as crude extracts.

This study clearly demonstrates that C. sativa leaf extracts, A. nilotica leaf extracts, and T. cordifolia stem extracts are potent molluscicides, and mortality caused by all the plant preparations was time and concentration dependent. There was a negative regression between LC values and exposure time. The toxicity of purified preparations of all three plants against L. acuminata is in the range of a potent molluscicide that is less than the threshold of 100 mg/L, set for a potential molluscicide by the World Health Organization (WHO) (Wei et al., 2002). Among all the organic solvent extracts, the higher toxicity of ethanol extracts of C. sativa leaf and A. nilotica leaf and acetone extracts of T. cordifolia stem indicate that the active molluscicidal components are soluble in ethanol and acetone, respectively, than other organic solvents.

Molluscicidal activity of C. sativa, A. nilotica, and T. cordifolia is due to the presence of β-caryophyllene, gallic acid, and berberine, as evident from individual toxicity and identification by TLC. Earlier, it was reported that β-caryophyllene is a potent molluscicide (Bedini et al., 2016). The pharmacological and biological effects of β-caryophyllene as an antiviral against Zika virus (Sobrinho et al., 2021); (Astani et al., 2011); antibacterial (Dahham et al., 2015; Moo et al., 2020); larvicidal (Huang et al., 2019; Hung et al., 2022); and anticancer (Dahham et al., 2015; Fidyt et al., 2016) have been noted.

Anti-inflammatory, antioxidant, antibacterial, anticancer, and antifungal activities of A. nilotica have been reported (Banso, 2009; Rather and Mohammad, 2015; Sadiq et al., 2017). Most of the biological activities of A. nilotica extracts are due to tannis (Sadiq et al., 2015). Different biological activities of T. cordifolia, such as antiviral infections, anticancer, antidiabetes, inflammation, neurological, and immunomodulatory effects in human beings, have been reported (Gupta and Sharma, 2011; Jagetia and Rao, 2006; Patel et al., 2009).

A comparison of the molluscicidal activity of β-caryophyllene, gallic acid, and berberine active components present in C. sativa, A. nilotica, and T. cordifolia, respectively, with synthetic molluscicides clearly demonstrates that these components are more potent against L. acuminata. Ninety-six-hour LC₅₀ of β-caryophyllene (3.47 mg/L), gallic acid (6.10 mg/L), and berberine (7.42 mg/L) are lower than those of synthetic molluscicides niclosamide (11.8 mg/L), carbaryl (14.40 mg/L), formothion (8.56 mg/L), and phorate (15.0 mg/L) (Singh et al., 2012).

Conclusion

The study findings suggest that the crude extracts, organic solvent extracts, and bioactive fractions of indigenous plants C. sativa, A. nilotica, and T. cordifolia have the potential to be used as molluscicides against the snail Lymnaea acuminata. This study discovered the potentiality of indigenous plants as cost-effective and eco-friendly molluscicides, and the results revealed that the molluscicides from plants are a better substitute for costly and harmful synthetic molluscicides. Further investigations are required to reveal the mode of action of these phytoconstituents responsible for the toxicity.

Footnotes

Authors' Contributions

V.K.S.: Supervision, conceptualization, methodology, software, reviewing, and editing. N.V.S.: Data curation, writing- original draft preparation, visualization, and investigation.

Author Disclosure Statement

No conflicting financial interests exist.

Funding Information

No funding was received for this article.

References

Adetunji

, Salawu

. Efficacy of ethanolic leaf extracts of Carica papaya and Terminalia catappa as molluscicides against the snail intermediate hosts of schistosomiasis. J Med Plants Res, 2010; 4(22):2348–2352; doi: 10.5897/JMPR10.468

Astani

, Reichling

, Schnitzler

. Screening for antiviral activities of isolated compounds from essential oils. Evid Based Complementary Altern Med, 2011; doi: 10.1093/ecam/nep187

Banso

. Phytochemical and antibacterial investigation of bark extracts of Acacia nilotica. J Med Plants Res, 2009; 3(2):082–085.

Bedini

, Flamini

, Cosci

. Cannabis sativa and Humulus lupulus essential oils as novel control tools against the invasive mosquito Aedes albopictus and fresh water snail Physella acuta. Ind Crops Prod, 2016; 85:318–323; doi: 10.1016/j.indcrop.2016.03.008

Dahham

, Tabana

, Iqbal

, et al. The anticancer, antioxidant and antimicrobial properties of the sesquiterpene β-caryophyllene from the essential oil of Aquilaria crassna. Molecules, 2015; 20(7):11808–11829; doi: 10.3390/molecules200711808

Echenique-Elizondo

, Amondarain

, Liron de Robles

. Fascioliasis: An exceptional cause of acute pancreatitis. J Pancreas, 2005; 6(1):36–39.

Fidyt

, Fiedorowicz

, Strządała

, et al. β-caryophyllene and β-caryophyllene oxide—Natural compounds of anticancer and analgesic properties. Cancer Med, 2016; 5(10):3007–3017; doi: 10.1002/cam4.816

Fürst

, Keiser

, Utzinger

. Global burden of human food-borne trematodiasis: A systematic review and meta-analysis. Lancet Infect Dis, 2012:12(3):210–221; doi: 10.1016/S1473-3099(11)70294-8

Gupta

, Sharma

. Ameliorative effects of Tinospora cordifolia root extract on histopathological and biochemical changes induced by aflatoxin-B1 in mice kidney. Toxicol Int, 2011; 18(2):94.

10.

Huang

, Lin

, Kuo

, et al. Phytochemical composition and larvicidal activity of essential oils from herbal plants. Planta, 2019; 250:59–68.

11.

Hung

, Pham

, Satyal

, et al. Strong larvicidal and molluscicidal activities of the (E)-β-caryophyllene-rich chemotype of callicarpa macrophylla vah leaf essential oil growing wild in vietnam. J Essent Oil Bear Plants, 2022; 25(3):666–680; doi: 10.1080/0972060X.2022.2104135

12.

Jagetia

, Rao

. Evaluation of the antineoplastic activity of guduchi (Tinospora cordifolia) in Ehrlich ascites carcinoma bearing mice. Biol Pharm Bull, 2006; 29(3):460–466; doi: 10.1248/bpb.29.460

13.

Jaiswal

, Singh

. Molluscicidal activity of Carica papaya and Areca catechu against the freshwater snail Lymnaea acuminata. Vet Parasitol, 2008; 152(3–4):264–270; doi: 10.1016/j.vetpar.2007.12.033

14.

Marston

, Hostettmann

. Review article number 6: Plant molluscicides. Phytochemistry, 1985; 24(4):639–652; doi: 10.1016/S0031-9422(00)84870-0

15.

Mas-Coma

, Bargues

, Valero

. Fascioliasis and other plant-borne trematode zoonoses. Int J Parasitol, 2005; 35(11–12):1255–1278; doi: 10.1016/j.ijpara.2005.07.010

16.

Mazeri

, Rydevik

, Handel

, et al. Estimation of the impact of Fasciola hepatica infection on time taken for UK beef cattle to reach slaughter weight. Sci Rep, 2017; 7(1):7319; doi: 10.1038/s41598-017-07396-1

17.

Moo

, Yang

, Osman

, et al. Antibacterial activity and mode of action of β-caryophyllene on Bacillus cereus. Pol J Microbiol, 2020; 69(1):49–54; doi: 10.3307/3/pjm-2020-007

18.

Patel

, Shah

, Goyal

. Antihyperglycemic, antihyperlipidemic and antioxidant effects of Dihar, a polyherbal ayurvedic formulation in streptozotocin induced diabetic rats. Indian J Exp Biol, 2009; 47:564–570; Available from: http://nopr.niscpr.res.in/handle/123456789/5043 [Last accessed: November 26, 2023).

19.

Portugaliza

, Balaso

, Descallar

, et al. Prevalence, risk factors, and spatial distribution of Fasciola in carabao and intermediate host in Baybay, Leyte, Philippines. Vet Parasitol Reg Stud Rep, 2019; 15:100261; doi: 10.1016/j.vprsr.2018.100261

20.

Rather

, Mohammad

. Acacia nilotica (L.): A review of its traditional uses, phytochemistry, and pharmacology. Sustain Chem Pharm, 2015; 2:12–30; doi: 10.1016/j.scp.2015.08.002

21.

Sadiq

, Hanpithakpong

, Tarning

, et al. Screening of phytochemicals and in vitro evaluation of antibacterial and antioxidant activities of leaves, pods and bark extracts of Acacia nilotica (L.) Del. Ind Crops Products, 2015; 77:873–882; doi: 10.1016/j.indcrop.2015.09.067

22.

Sadiq

, Tarning

, Aye Cho

, et al. Antibacterial activities and possible modes of action of Acacia nilotica (L.) Del. against multidrug-resistant Escherichia coli and Salmonella. Molecules, 2017; 22(1):47; doi: 10.3390/molecules22010047

23.

Singh

, Singh

. Characterization of the molluscicidal activity of Bauhinia variegata and Mimusops elengi plant extracts against the fasciola vector Lymnaea acuminata. Rev Inst Med Trop São Paulo, 2012; 54:135–140; doi10.1590/S0036-46652012000300004

24.

Singh

, Singh

. Molluscicidal activity of cannabis sativa, acacia niloticaand tinospora cordifolia extracts against vector snail lymnaea acuminata. Trends Appl Sci Res, 2023; 18(2):41–49; doi10.17311/tasr.2023.41.49

25.

Singh

, Singh

. Characterization of allicin as a molluscicidal agent in Allium sativum (Garlic). Biol Agric Hortic, 1995; 12(2):119–131; doi: 10.1080/01448765.1995.9754732

26.

Sobrinho

, de Morais

, Marinho

, et al. Antiviral activity on the Zika virus and larvicidal activity on the Aedes spp. of Lippia alba essential oil and β-caryophyllene. Ind Crops Products, 2021; 162:113281; doi: 10.1016/j.indcrop.2021.113281

27.

Wei

, Xu

, Liu

, et al. Toxicology of a potential molluscicide derived from the plant Solanum xanthocarpum: A preliminary study. Ann Trop MedParasitol, 2002; 96(3):325–331; doi: 10.1179/000349802125000727

28.

Yattoo

, Dar

, Jan

, et al. Human fascioliasis: Report of two cases from Kashmir Valley. J Clin Exp Hepatol, 2021; 11(6):747–750; doi: 10.1016/j.jceh.2020.12.010