A Comprehensive Review of the Pharmacology, Chemistry, Traditional Uses and Quality Control of Star Anise (Illicium verum Hook. F.): An Aromatic Medicinal Plant.

Illicium verum Hook. F., also known as star anise, is one of the most important plants of the genus Anise in the family Magnoliaceae. I. verum not only has the functions of warming Yang, dispersing cold, regulating Qi and relieving pain but can also be used as a condiment to increase flavor as well as reconcile and remove fish smells. Currently, 201 chemical constituents have been identified from star anise; among these, star anise oil and shikimic acid are the two most widely used and studied chemical components in star anise, with the oil accounting for a large proportion of the total. This review integrates, classifies and updates studies related to the botany, pharmacology, phytochemistry, traditional and modern uses and quality control of star anise, with a special reference to its phytochemical composition and pharmacological activity. It will provide a reference for further research on this important medicinal plant. In addition, the broad applications and research profiles of star anise essential oil and shikimic acid are highlighted. Our review indicates that the research prospects regarding star anise are very broad and worthy of further investigation.

Antidiabetic Effect of Star Anise (Illicium verum) in Streptozotocin-induced Diabetic Rats.

AIM: The current study aimed to evaluate the antidiabetic potential of Illicium verum fruits. BACKGROUND: Illicium verum fruits are frequently used by the Moroccan population in the treatment of diabetes. METHODS: The antihyperglycemic effect of the aqueous extract of Illicium verum fruits (AEIVF) in rats was assessed. The effects of AEIVF (20 mg/kg) on glycemia and lipid profile as well as its phytochemical and antioxidant properties were evaluated. RESULTS: In normal and diabetic rats, AEIVF reduced blood glucose levels 6 hours after administration. Furthermore, after 7 days of treatment, glycemia was lowered in diabetic rats, and this extract exhibited an antioxidant activity. CONCLUSION: The study shows that Illicium verum possesses a potent antidiabetic activity. In addition, the toxicity of AEIVF was evaluated and the LD50 value was found to be greater than the 2 g/kg dose.

Diverse prenylated C6-C3 derivatives from the stems and branches of Illicium ternstroemioides A. C. Smith with anti-inflammatory and antiviral activities.

Fifteen unreported prenylated C6-C3 derivatives (1-15) were isolated from the stems and branches of Illicium ternstroemioides A. C. Smith, including one bis-prenylated C6-C3 derivative (1), three prenylated C6-C3 derivative-shikimic acid ester hybrids (2-4) and 11 prenylated C6-C3 monomers (5-15). The structures of compounds 1-15 were elucidated by spectroscopic analysis (UV, IR, 1D and 2D NMR, and HRESIMS). The absolute configurations of the compounds were determined using electronic circular dichroism (ECD), induced circular dichroism (ICD), and the modified Mosher's method. Among the isolates, compounds 11, 12, and 15 exhibited significant anti-inflammatory activities by inhibiting the nitric oxide with IC50 values ranging from 1.89 to 24.83 µM in lipopolysaccharide-stimulated murine RAW 264.7 macrophages and murine BV2 microglial cells; compounds 2, 3, and 7 exhibited antiviral activitives against Coxsackievirus B3 with an IC50 value of 33.3, 25.9, and 27.8 µM, respectively.

Simultaneous determination of three curative flavonoids and neurotoxic anisatin in star anise fruits by ultra-high performance liquid chromatography-tandem mass spectrometry.

Chinese star anise is an anise-scented star-shaped fruit of the evergreen Illicium verum tree. It is generally used as a spice and folk medicine for treating digestive illness. Chinese star anise may be adulterated with Japanese star anise (fruit of Illicium anisatum tree with similar morphology to the former), containing high content of toxic anisatin leading to severe physical disorders. In this work, a UHPLC-MS/MS method has been developed to determine three quality markers (quercetin, kaempferol, and luteolin) and toxicity marker (anisatin) of star anise using quercetin-d3, an exogenous heavy isotopic compound, as internal standard. Chromatographic separation of quality and toxicity marker compounds was achieved using a T3 column within two minutes. The method was successfully developed, validated, and used to quantify the multiple marker compounds in both botanical reference materials and star anise samples. This method can be used for quality control and assessment of Chinese star anise products.

Toxicological and molecular adverse effect of Illicium verum fruit constituents toward Bradysia procera.

BACKGROUND: Bradysia procera, a ginseng stem fungus gnat, is one of the most serious insect pests of Korean ginseng (Panax ginseng), causing significant damage to plant growth. The goal of this study was to determine the toxicity and mechanism of action of phenylpropanoids (trans-anethole and estragole) isolated from the methanol extract and hydrodistillate of Illicium verum fruit against third-instar larvae and eggs of Bradysia procera. RESULTS: The filter-paper mortality bioassay revealed that estragole [median lethal concentration (LC50 ) = 4.68 g/cm2 ] has a significant fumigant effect, followed by trans-anethole (LC50 = 43.92 g/cm2 ). However, estragole had the lowest toxic effect when compared to commercially available insecticides. After 7 days, estragole and trans-anethole at 75 g/cm2 inhibited egg hatchability up to 97% and 93%, respectively. At 0.09 g/cm2 , insecticides had an inhibitory effect on egg-hatching ability ranging from 88% to 94%. Furthermore, in both closed and open containers, these active constituents were able to consistently induce vapor-phased toxicity. Both estragole and trans-anethole have the ability to inhibit acetylcholinesterase (AChE), which is involved in neurotransmitter function. However, the active constituent estragole from I. verum fruit acted as a potent AChE inhibitor and had a slightly lower effect on cyclic adenosine monophosphate (AMP) than octopamine alone. CONCLUSION: This finding suggests that estragole may influence Bradysia procera neurotransmitter function via both the AChE and octopaminergic receptors. More research is needed to demonstrate the potential applications of I. verum fruit-derived products as potential larvicides and ovicides for Bradysia procera population control. © 2022 Society of Chemical Industry.

Geranyl phenyl ethers from Illicium micranthum and their anti-HBV activity.

Fourteen new geranyl phenyl ethers (1-14) along with three known compounds (15-17) were isolated from Illicium micranthum, and their structures were elucidated by comprehensive spectroscopic methods. Illimicranins A-H (1-8) were characterized as geranyl vanillin ethers, while 9 and 10 were dimethyl acetal derivatives. Illimicranins I and J (11 and 12) were rare geranyl isoeugenol ethers. Illimicranins K and L (13 and 14) represented the first example of geranyl guaiacylacetone ether and geranyl zingerone ether, respectively. Compounds 1, 2 and 15 exhibited anti-HBV (hepatitis B virus) activity against HBsAg (hepatitis B surface antigen) and HBeAg (hepatitis B e antigen) secretion, and HBV DNA replication.

Illicium verum (Star Anise) and Trans-Anethole as Valuable Raw Materials for Medicinal and Cosmetic Applications.

Illicium verum Hook f. (star anise) is considered an important species in Traditional Chinese Medicine and is also used in contemporary medicine in East Asian countries. It occurs in natural habitats in southeastern parts of China and Vietnam, and is cultivated in various regions in China. The raw materials-Anisi stellati fructus and Anisi stellati aetheroleum obtained from this species exhibit expectorant and spasmolytic activities. The European Pharmacopoeia (4th edition) indicates that these raw materials have been used in allopathy since 2002. The biological activities of the above-mentioned raw materials are determined by the presence of valuable secondary metabolites such as monoterpenoids, sesquiterpenoids, phenylpropanoids, and flavonoids. Recent pharmacological studies on fruit extracts and the essential oil of this species have confirmed their antibacterial, antifungal, anti-inflammatory, and antioxidant activities and thus their medicinal and cosmetic value. The aim of this review was to examine the progress of phytochemical and pharmacological studies that focused on possible cosmetic applications. In addition to fruit extracts and essential oil, the current consensus on the safety of trans-anethole, which is the main compound of essential oil used in cosmetology, is underlined here.

seco-Prezizanne Sesquiterpenes and Prenylated C6-C3 Compounds from the Fruits of Illicium lanceolatum A. C. Smith.

Two new seco-prezizaane-type sesquiterpenes, 2ß-hydroxy-6-deoxyneoanisatin (1) and 3,4-anhydro-2-oxo-1α-hydroxy-6-deoxyneoanisatin (2), and two new prenylated C6 -C3 compounds, illilanceofunones A (3) and B (4), were obtained from the fruits of Illicium lanceolatum, along with four known prenylated C6 -C3 compounds (5-8). Their structures were proposed through HR-ESI-MS, 1 H, 13 C, and 2D NMR data interpretation. Moreover, the absolute configuration of 1 and 2 were further assigned by single-crystal X-ray diffraction analysis and electronic circular dichroism (ECD) calculations, respectively. Illihenryipyranol A (6) exhibited neuroprotective activity against MPP+ -induced PC12 cell damage in a dose-dependent manner.

Preparation, characterization, and evaluation of HP-ß-CD inclusion complex with alcohol extractives from star anise.

The active compounds in star anise alcohol extractives (SAAE) have potent bioactivity. However, their poor solubility and stability limit their applications. In this study, SAAE/hydroxypropyl-ß-cyclodextrin (HP-ß-CD) inclusion complexes were prepared as a strategy to overcome the abovementioned disadvantages. The phase solubility results indicated that the solubility of the inclusion complex was enhanced. Complexation was confirmed by complementary methods, including Fourier-transform infrared spectroscopy, nuclear magnetic resonance spectroscopy, scanning electron microscopy, and transmission electron microscopy, which proved to be extremely insightful for studying the inclusion formation phenomenon between SAAE and HP-ß-CD. Despite there being no apparent improvements in the antioxidant capacity and antimicrobial activity, the results of the stability studies presented higher thermal, volatile, and photostability after encapsulation. Further, molecular modeling was used to investigate the factors influencing complex formation and provide the most stable molecular conformation. Thus, based on the obtained results, this study strongly demonstrates the potential of the SAAE/HP-ß-CD inclusion complex in the food industry.

Comparison of the Phytochemical Composition and Antibacterial Activities of the Various Extracts from Leaves and Twigs of Illicium verum.

Previous studies have revealed the numerous biological activities of the fruits of Illicium verum; however, the activities of its leaves and twigs have remained undiscovered. The study aimed to investigate the phytochemical components and antibacterial activity of the various extracts from the leaves and twigs of Illicium verum. The herbal extracts were prepared by supercritical CO2 extraction (SFE) and 95% ethanol extraction, followed by partition extraction based on solvent polarity. Analysis of antimicrobial activity was conducted through the usage of nine clinical antibiotic- resistant isolates, including Staphylococcus aureus, Pseudomonas aeruginosa and Acinetobacter baumannii. Among the tested samples, the SFE extracts exhibited broader and stronger antibacterial activities against the test strains, with a range of MIC between 0.1-4.0 mg/mL and MBC between 0.2-4.5 mg/mL. Observations made through scanning electron microscopy revealed potential mechanism of the antimicrobial activities involved disruption of membrane integrity of the test pathogens. Evaluation of the chemical composition by gas chromatography-mass spectrometry indicated the presence of anethole, anisyl aldehyde, anisyl acetone and anisyl alcohol within the SFE extracts, demonstrating significant correlations with the antibacterial activities observed. Therefore, the leaves and twigs of Illicium verum hold great potential in being developed as new natural antibacterial agents.

Structure elucidation and anti-inflammatory mechanism of difengpienol C, a new neolignan isolated from Illicium difengpi.

Illicium difengpi is well-known as its stem barks that have been widely used in the Traditional Chinese Medicine (TCM) for therapy rheumatoid arthritis and traumatic injury. To comprehensive utilization of resources, the phytochemical investigation on the branches and leaves of this plant was carried out, which led to the isolation of an undescribed neolignan along with three known lignans. Their structures were elucidated on the basis of extensive spectroscopic data and the new compound was elucidated as a neolignan possessing a dihydropyran ring formed by a unique conjugation way and named difengpienol C. Difengpienol C showed the strongest anti-inflammatory activity in lipopolysaccharide (LPS)-stimulated RAW264.7 cells, which powerfully inhibited nitric oxide (NO), interleukin 6 (IL-6), tumor necrosis factor α (TNF-α) production and suppressed the mRNA transcription of inducible nitric oxide synthase (iNOS), IL-6 and TNF-α. Besides, difengpienol C blocked the activation of TLR4/MyD88/NF-κB signaling pathway. Therefore, difengpienol C might be a potent agent for anti-inflammatory drug development, and the non-traditional medicinal parts of Illicium difengpi can be identified as the source of natural anti-inflammatory molecules.

Effects of star anise (Illicium verum Hook.f.) essential oil administration under three different dietary energy levels on growth performance, nutrient, and energy utilization in broilers.

This experiment was conducted to investigate the effect of star anise essential oil (SAO) supplementation in diets with different energy levels on growth performance, nutrient metabolic efficiency of broilers. One hundred and ninety-two Arbor Acres male broiler chicks at 28 days of age were divided into a 3 × 4 factorial arrangement design with three dietary energy levels (13.41, 12.82, 12.23 MJ/kg) and 4 levels of SAO supplementation (0, 200, 400, and 600 mg/kg of diet). Dietary supplementation with SAO increased (p < .05) apparent metabolic efficiency of CP, EE, GE, and all of the amino acids except Trp. Significant interactions were noted between energy level and SAO administration for metabolic efficiency of CP, all of the amino acids except Cys. Inclusion of SAO enhanced apparent nutrient metabolic efficiency of broilers in a dose-dependent manner, birds supplemented with 400 mg/kg of SAO in high-energy diets appeared to contain highest nutrient metabolic efficiency, moreover, the metabolic efficiency of nutrients in low-energy diets along with 200 or 400 mg/kg of SAO was similar with that in high-energy diets without SAO, which indicated that the SAO might ameliorate the negative effects of reduced dietary metabolic energy on nutrient utilization in broilers.

Theoretical and experimental investigation of sodium alginate composite films containing star anise ethanol extract/hydroxypropyl-ß-cyclodextrin inclusion complex.

The aim of this study is to prepare composite films incorporated with star anise ethanol extract (SAEE)/hydroxypropyl-ß-cyclodextrin (HP-ß-CD) inclusion complex. The effects of sodium alginate concentration on mechanical properties of films are tested. Sodium alginate, SAEE, and SAEE/HP-ß-CD inclusion complex-based composite films are characterized in terms of UV-visible spectroscopy, microstructure characterizations, including transmission electron microscopy, scanning electronic microscopy, Fourier transform infrared, and 1 H NMR analysis, and molecular modeling calculations. With various stoichiometries, the complexes of sodium alginate/SAEE/HP-ß-CD are compared through both theoretical and experimental analyses. Molecular simulations are applied to predict the possible orientations of SAEE inside the HP-ß-CD cavity and the optimal stoichiometry of the complex formation. According to the simulation, the system of sodium alginate/SAEE (or SAEE/HP-ß-CD inclusion complex) in a 3:1 stoichiometry reaches the lowest total energy and achieves a balance in complex system. In addition, the composite films can maintain high-content vitamin C and reduce weight loss rate of fresh-cut Chinese yam. In conclusion, coinciding with the experimental results, the molecular modeling successfully calculates the reasonable molecular structure and molecular behavior of sodium alginate/SAEE/HP-ß-CD inclusion complex. The composite films in this study have the potential to be used for food packaging applications. PRACTICAL APPLICATION: In this paper, we present composite films incorporated with star anise ethanol extract (SAEE)/hydroxypropyl-ß-cyclodextrin (HP-ß-CD) inclusion complex for the use of fresh-cut Chinese yam preservation. The present study demonstrates a successful application of molecular modeling to predict the geometry of the final complex. It can serve as a powerful tool to calculate the energy of association between inclusion complex and sodium alginate molecules.

Improvement of the stabilities and antioxidant activities of polyphenols from the leaves of Chinese star anise (Illicium verum Hook. f.) using ß-cyclodextrin-based metal-organic frameworks.

BACKGROUND: Polyphenols extracted from plants are usually highly unstable and rapidly transformed into various reaction products during food and drug processing, thus limiting their applications. To improve the stability and solubility of polyphenols from the leaves of Chinese star anise (Illicium verum Hook. f.), and hence to expand their application to food and medicine, the extracted anise leaf polyphenols (ALPs) were microencapsulated using ß-cyclodextrin (ß-CD) and cyclodextrin-based metal-organic frameworks (ß-CD-MOFs). RESULTS: The optimum inclusion rate of ALP/ß-CD-MOFs was 97.80% at a core-wall ratio of 1:10. Meanwhile, the stabilities, solubilities and antioxidant activities of the polyphenols before and after inclusion were compared. The results showed both the stabilities and solubilities of ALP/ß-CD-MOFs were significantly improved compared with those of ALPs and ALP/ß-CD, suggesting the potential of ß-CD-MOFs as newer and better carriers than ß-CD for polyphenols in food industry applications. The free radical (including superoxide, hydroxyl and DPPH radicals) scavenging activities were also improved by microencapsulation. Superoxide radical scavenging reaction also showed slow-release property of ALP/ß-CD-MOFs. The formation of the inclusion complex was further confirmed using Fourier transform infrared spectral characterization. CONCLUSIONS: Microencapsulation with ß-CD-MOFs could expand the application scope of ALPs, and it is more effective than encapsulation with ß-CD. This is important for a better understanding and application of this useful traditional Chinese plant. As a new material with high efficiency and edibility, ß-CD-MOFs are not limited to the chemical field, but also have potential in new areas of food, medicine and healthcare products. © 2020 Society of Chemical Industry.

Effects of star anise (Illicium verum Hook. f) and its extractions on carcass traits, relative organ weight, intestinal development, and meat quality of broiler chickens.

Star anise (Illicium verum Hook. f) has been used as spice and herbal medicine for many years with lacking researches on evaluating its application for improving relative organ weight, intestinal development, and quality of animal products. The experiment was conducted to investigate the effects of star anise, its essential oil and leavings on carcass traits, relative organ weight, intestinal development, and meat quality of broiler chickens. A total of 384 broilers were randomly assigned to 4 treatments with 8 replicates of 12 birds each. The 4 dietary treatments were basal diet (Control), basal diet supplemented with 5 g/kg star anise (Star anise), basal diet supplemented with 0.22 g/kg essential oil (Essential oil), and basal diet supplemented with 5 g/kg leavings (Leavings). The concentration of main active components in the experimental diets was 0.204 g trans-anethole/kg of diet. All birds were fed a starter diet (0-21 d) and a grower diet (22-42 d). Birds supplemented with star anise and essential oil had a greater (P < 0.05) final body weight (BW) than control birds and those supplemented with leavings. However, the carcass yield, half chamber rate, eviscerated rate, and percentages of breast muscle and thigh muscle in birds were similar (P > 0.05) among all treatments. Birds supplemented with star anise and essential oil had higher (P = 0.010) relative weight of thymus than those in control and leavings groups. Essential oil supplemented groups appeared to contain the highest (P < 0.05) villus height in ileum and villus height/crypt depth ratio in ileum and jejunum among all the groups. Meanwhile, among all the groups, breast muscles of essential oil-supplemented groups appeared to contain the lowest (P = 0.012) boiling loss but highest (P < 0.001) concentration of inosinic acid (inosine 5'-monophosphate, IMP). In conclusion, dietary supplementation of 5 g/kg star anise and 0.22 g/kg essential oil improved BW, relative organ weight, and intestinal development, and 0.22 g/kg essential oil can also increase the concentration of IMP but decrease the boiling loss in breast muscles of broilers; however, 5 g/kg leavings had no effect.

Effects of star anise (Illicium verum Hook.f) oil on the nuclear factor E2-related factor 2 signaling pathway of chickens during subclinical Escherichia coli challenge.

We characterized the mechanism underlying star anise (Illicium verum Hook.f) oil (SAO)-mediated antioxidant status during subclinical Escherichia coli (E. coli) challenge. A total of 512 male birds (White Leghorn) at 30 wk of age with similar body weight (2.14 ± 0.02 kg) were randomly divided into 2 groups with 1 group being orally challenged with E. coli (every other day from day 15 to day 27) during the experiment. Each group of birds was then randomly allocated to dietary treatment of SAO supplementation at 0, 200, 400, or 600 mg/kg of basal diet (8 replicate cages during each treatment). The treatments were arranged a 4 × 2 factorial arrangement. The experiment comprised 1 wk of adaptation and 3 wks of data collection. There was no interaction (P > 0.05) between SAO supplementation and E. coli challenge for final body weight and average daily feed intake of birds. However, E. coli challenge resulted in a significant decrease (P < 0.001) in final body weight of birds as compared with unchallenged birds. There were interactions between SAO supplementation and E. coli challenge for the activity of glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) concentration in serum and for the activity of GSH-Px in the liver of birds. Supplementation of SAO enhanced the activities of antioxidant enzymes but decreased the MDA content in the serum and liver of birds, and it also enhanced the expression of genes including superoxide dismutase, catalase, and nuclear factor E2-related factor 2 (Nrf2) in the liver of the birds. Meanwhile, supplementation of SAO can also reduce E. coli challenge-induced oxidative stress in the serum and liver of birds, and the efficacy of SAO in birds during subclinical E. coli challenge is dose-dependent. In conclusion, the enhancement of antioxidant capacity by star anise or its effective compounds is through upregulation of Nrf2 signaling pathway. The optimum supplementation dose of SAO for protecting birds against E. coli challenge is 400 mg/kg.

Antiviral activity of Illicium verum Hook. f. extracts against grouper iridovirus infection.

Grouper iridovirus causes high mortality rates in cultured groupers, and effective treatment for grouper iridovirus infection is urgently required. Illicium verum Hook. f. is a well-known medicinal plant with a variety of biological activities. The aim of this study was to analyse the use of I. verum extracts to treat grouper iridovirus infection. The safe working concentration of each I. verum extract was identified both in vitro and in vivo as follows: I. verum aqueous extract (IVAE) ≤ 500 µg/ml; I. verum ethanol extract (IVEE) ≤ 250 µg/ml; shikimic acid (SKA) ≤ 250 µg/ml; trans-anethole (TAT) ≤ 800 µg/ml; 3,4-dihydroxybenzoic acid (DDBA) ≤ 400 µg/ml; and quercetin (QCE) ≤ 50 µg/ml. The inhibitory activity of each I. verum extract against grouper iridovirus infection was analysed using aptamer (Q2)-based fluorescent molecular probe (Q2-AFMP) and RT-qPCR. All of the I. verum extracts displayed dose-dependent antiviral activities against grouper iridovirus. Based on the achieved per cent inhibition, IVAE, IVEE, DDBA and QCE were associated with the greatest antiviral activity (all > 90%). Together, our results indicate that I. verum extracts have effective antiviral properties, making it an excellent potential source material for the development of effective treatment for grouper iridovirus infection.

Star anise (Illicium verum): Chemical compounds, antiviral properties, and clinical relevance.

Medicinal herbs are one of the imperative sources of drugs all over the world. Star anise (Illicium verum), an evergreen, medium-sized tree with star-shaped fruit, is an important herb with wide distribution throughout southwestern parts of the Asian continent. Besides its use as spice in culinary, star anise is one of the vital ingredients of the Chinese medicinal herbs and is widely known for its antiviral effects. It is also the source of the precursor molecule, shikimic acid, which is used in the manufacture of oseltamivir (Tamiflu®), an antiviral medication for influenza A and influenza B. Besides, several other molecules with numerous biological benefits including the antiviral effects have been reported from the same plant. Except the antiviral potential, star anise possesses a number of other potentials such as antioxidant, antimicrobial, antifungal, anthelmintic, insecticidal, secretolytic, antinociceptive, anti-inflammatory, gastroprotective, sedative properties, expectorant and spasmolytic, and estrogenic effects. This review aimed to integrate the information on the customary attributes of the plant star anise with a specific prominence on its antiviral properties and the phytochemical constituents along with its clinical aptness.

Phytochemical constituents of the pericarps of Illicium difengpi and their anti-inflammatory activity.

Phytochemical investigation on the pericarps of Illicium difengpi lead to the isolation and structure elucidation of a new sesquiterpene, sesquicaranoic acid C (1), a new neolignan, difengpiol C (2), and 10 known compounds. The structures and absolute configurations of two new compounds were determined by a combination of NMR and CD spectroscopic analyses. All isolates were evaluated for their inhibitory effects on lipopolysaccharide (LPS)-induced nitric oxide (NO) production in RAW264.7 cells.

In Vitro Anthelmintic Activity of Three Tropical Plant Extracts on Haemonchus contortus.

AIM: To evaluate the anthelmintic activity of Anacardium occidentale shell, Illicium verum fruit, and Artocarpus heterophyllus seed to substantiate their traditional use against helminths. MATERIALS AND METHODS: In vitro anthelmintic activity of hydroalcoholic extracts of three plants was evaluated against eggs, infective larvae (L3), adult, and lactate dehydrogenase (LDH) of Haemonchus contortus of naturally infected sheep. RESULTS: The three extracts exhibited significant (P < 0.001) dose-dependent anthelmintic responses by inhibiting egg hatching and causing paralysis of larvae and mortality of worms. The extracts were most effective on egg and adult stage of H. contortus than on L3 stage at a higher concentration of 6 mg/mL. Anacardium occidentale shell exhibited maximum activity with 100% paralysis of L3 larvae. Probit analysis revealed that the extracts of A. Occidentale shell induced 50% egg hatch inhibition (LD50 = 0.0255 mg/mL), larval paralysis (LD50 = 0.196 mg/mL), and adult worm mortality (LD50 = 1.0365 mg/mL) at a lower concentration (LD50) compared with those of I. verum fruit and A. heterophyllus seed extracts. Further, all extracts significantly (P < 0.01) inhibited the LDH activity catalyzing the oxidation of lactate in adult H. contortus, with maximum level of inhibition caused by A. occidentale shell extract. CONCLUSION: Phytochemical screening of the extracts revealed the presence of alkaloids, flavonoids, tannins, saponins, and amino acids that could be responsible for the anthelmintic effects noticed. The results warrant further in vivo evaluation of these plants for potential use as anthelmintic agents.