Evaluation of anthelmintic efficacy of ethanolic leaf extract of Juglans regia L. on Ascaridia galli: a comprehensive in vitro and in vivo study.

Anthelmintic resistance in livestock animals has been spreading across the world in prevalence and severity. As a result, researchers are exploring alternative strategies to combat this issue, and one promising avenue is the utilization of medicinal plants. This study aims to investigate the anthelmintic efficacy of the crude ethanolic extract (CEE) derived from the leaves of Juglans regia against one of the most detrimental nematode parasites affecting poultry, namely Ascaridia galli (A. galli). For the in vitro studies, adult A. galli worms were collected from the naturally infected chickens and the efficacy of CEE was measured at the concentration of 25, 50, and 100 mg/ml using adult worm motility inhibition (WMI) assay. In addition, levamisole (0.55 mg/ml) was used as the positive control. Likewise, Phosphate buffered saline (PBS) was used as the negative control. For the in vivo studies, CEE of J.regia at the doses of 500, 1000, and 2000 mg/kg were evaluated in chickens experimentally infected with A. galli. The anthelmintic efficacy was monitored using faecal egg count reduction (FECR) and worm count reduction (WCR) assays. In vitro studies revealed significant (P < 0.001) anthelmintic effects of CEE of J.regia on the motility of A. galli worms at different hours post-exposure. At the concentration of 100 mg/ml, CEE resulted in 96.5% inhibition of worm motility at 24 h post-exposure. While the synthetic anthelmintic drug, levamisole caused the highest inhibition of worm motility (100%) at the same time period. The in vivo anthelmintic activity of CEE of J. regia demonstrated a maximum effect on day 14 post-treatment by inducing 67.28% FECR and 65.03% WCR. We observed no significant difference (P > 0.05) in worm counts between the negative control group and the chickens treated with CEE at the dosage of 500 mg/kg. Together, the results of the present study suggest that CEE of J. regia leaves possess anthelmintic properties and could be a potential source of novel anthelmintic compounds for controlling helminth parasites.

In vitro assessment of the anthelmintic activity of copper oxide and zinc oxide nanoparticles on egg and adult stages of Fasciola hepatica: evidence on oxidative stress biomarkers, and DNA damage.

OBJECTIVES: Fasciolosis is of significant economic and public health importance worldwide. The lack of a successful vaccine and emerging resistance in flukes to the drug of choice, triclabendazole, has initiated the search for alternative approaches. In recent years, metallic nanoparticles have been extensively investigated for their anthelmintic effects. This study investigates the in vitro anthelmintic activity of copper oxide and zinc oxide nanoparticles against Fasciola hepatica. METHODS: The in vitro study was based on egg hatchability test (EHA), adult motility inhibition tests, DNA damage, ROS levels, as well as several biomarkers of oxidative stress, including glutathione peroxidase (GSH) and glutathione S-transferase (GST), superoxide dismutase (SOD) and malondialdehyde (MDA). For this purpose, different concentrations of copper oxide nanoparticles (CuO-NPs) and Zinc oxide nanoparticles (ZnO-NPs) (1, 4, 8, 12, and 16 ppm) were used to evaluate the anthelmintic effect on different life stages, including egg and adults of Fasciola hepatica, over 24 h. RESULTS: In vitro treatment of F. hepatica worms with both CuO-NPs and ZnO-NPs could significantly increase ROS production and oxidative stress induction (decreased SOD, GST and GSH and increased MDA) compared to control group. CONCLUSIONS: Based on the results, it seems that CuO-NPs and ZnO-NPs may be effective in the control and treatment of F. hepatica infection. Further research is needed to investigate their potential for in vivo use in the treatment of parasitic infections.

Structure-based discovery of potent myosin inhibitors to guide antiparasite drug development.

Monogenea, a prevalent parasite in aquaculture, poses significant threats to the industry, leading to substantial losses. Current preventive measures have proven insufficient, necessitating the development of novel and effective anti-parasitic drugs. In this investigation, we obtained the full-length myosin cDNA sequence by analyzing three-generation transcriptome data, revealing a 5817-base sequence encoding 1938 amino acids. Subsequently, we modeled and analyzed the characteristics of the secondary and tertiary of myosin, pinpointing the crucial functional region within the motor domain (amino acids 1-768). The prokaryotic expression of this domain yielded a protein of 87.44 kDa, confirmed as myosin by Western Blotting. Molecular docking identified ASN439 as the key amino acid residue involved in arctigenin and myosin binding, a result corroborated by site-directed mutagenesis, affirming the active cavity of this interaction. Chalcone and shikonin were chosen from a virtual sieve of molecular library of natural drugs based on the active cavity. Chalcone and shikonin exhibited EC50 values of 1.085 mg/L and 0.371 mg/L, respectively, with corresponding IC50 values for myosin of 0.44 mM and 0.14 mM. Given its superior activity and structure, shikonin was selected for further optimization of drug molecule design, culminating in the discovery of 1,4-naphthoquinone as a potent antiparasitic agent. This compound demonstrated an EC50 of 0.047 mg/L, LC50 of 0.23 mg/L, and a TI index of 4.893. These findings collectively highlight the potential of shikonin and 1,4-naphthoquinone as alternative compounds to control Gyrodactylus infections. Further optimization of medicinal chemistry holds promise for the development of more potent 1,4-naphthoquinone analogues, offering prospects for future anthelmintic control through combinatorial or replacement strategies.

In Vitro anthelmintic efficacy of medicinal plant essential oils against Marshallagia marshalli: Evidence on oxidative/nitrosative stress biomarkers, DNA damage, and egg hatchability.

One of the major public health problems is drug resistance in parasitic diseases. It is therefore important to find new active ingredients to combat parasites. Herbal products such as essential oils (EOs) may show promise in treating infections caused by gastrointestinal nematodes (GINs). This study investigated the in vitro anthelmintic activity of the EOs of Lavandula angustifolia and Quercus infectoria against Marshallagia marshalli. The in vitro study was based on an egg hatch test (EHT), adult and larval motility inhibition tests, DNA damage, and several biomarkers of oxidative/nitrosative stress, including superoxide dismutase [SOD], catalase [CAT], and glutathione peroxidase [GSH -Px], protein carbonylation [PCO], malondialdehyde [MDA], total antioxidant status [TAS], and nitric oxide levels [NO]. Different concentrations of Lavandula angustifolia and Quercus infectoria EOs (1, 5, 10, 25 and 50 mg/ml) were used to determine the anthelmintic effect on three stages of the life cycle of M. marshalli, i.e. eggs, larvae and adult parasites, for 24 hr. The results showed that EOs of L. angustifolia and Q. infectoria play an important role as anthelmintics. These essential oils significantly reduced the egg hatching and motility of larval and adult worms. This anthelmintic effect is dependent on concentration and time. Furthermore, the EOs of L. angustifolia and Q. infectoria caused oxidative/nitrosative stress (reduced SOD, GSH-Px and CAT and increased MDA, PCO and NO) and DNA damage, thereby providing significant antihelminthic effects. Based on the results, it seems that the EOs extracted from L. angustifolia and Q. infectoria may be effective in the control and treatment of M. marshalli infections. Further research is needed to investigate their potential for in vivo use in the treatment of parasitic infections.

Structural basis for the recognition of anthelmintic activity of bioactive metabolite in watery rose apple leaf through in silico investigation.

This research is part of the research umbrella regarding the use of watery rose apple leaf [Eugenia aqueum (Burm. F) Alston] in the health sector. Infectious diseases of worms, are still a health problem with a high burden based on the incidence and mortality rates in Southeast Asian countries. Therefore, we tried to create a 3D structure of vital receptors of worms including Nicotinic Cholinergic Receptor (NCR), Acetylcholinesterase Enzymes (AE), and Phosphorylase Enzymes (PE) using homology modeling method through the SWISS-MODEL webserver, then conducted a molecular dynamics simulations using active metabolite of watery rose apple leaf i.e. 2',4'­dihydroxy­6'­methoxy­3',5'­dimethylchalcone which was then compared with Piperazine Citrate and Pyrantel Pamoate which are anthelmintic drugs on the market. Based on the results of the study, the structure of the three receptors with high resolution was successfully obtained which was characterized by a good Ramachandran value, which was above 90%. Then the ligand structure was successfully modeled in a three-dimensional model and optimized geometrically using GaussView 5.0.8 and Gaussian09 software. In further, Root Mean Square Deviation (RMSD) and Root Mean Square Fluctuation (RMSF) analysis of molecular dynamics simulations ligand against the three receptors, compound 2',4'­dihydroxy­6'­methoxy­3',5'­dimethylchalcone showed better stability than Piperazine Citrate and Pyrantel Pamoate at 50 ns simulation, although from the calculation of binding free energy Molecular Mechanics Poisson-Boltzmann Surface Area (MM/PBSA) is not better than the two comparisons. It can be concluded that the compound 2',4'­dihydroxy­6'­methoxy­3',5'­dimethylchalcone from watery rose apple leaf showed potential as anthelmintic candidates.Communicated by Ramaswamy H. Sarma.

Benzimidazoles Containing Piperazine Skeleton at C-2 Position as Promising Tubulin Modulators with Anthelmintic and Antineoplastic Activity.

Benzimidazole anthelmintic drugs hold promise for repurposing as cancer treatments due to their interference with tubulin polymerization and depolymerization, manifesting anticancer properties. We explored the potential of benzimidazole compounds with a piperazine fragment at C-2 as tubulin-targeting agents. In particular, we assessed their anthelmintic activity against isolated Trichinella spiralis muscle larvae and their effects on glioblastoma (U-87 MG) and breast cancer (MDA-MB-231) cell lines. Compound 7c demonstrated exceptional anthelmintic efficacy, achieving a 92.7% reduction in parasite activity at 100 µg/mL after 48 hours. In vitro cytotoxicity analysis of MDA-MB 231 and U87 MG cell lines showed that derivatives 7b, 7d, and 7c displayed lower IC50 values compared to albendazole (ABZ), the control. These piperazine benzimidazoles effectively reduced cell migration in both cell lines, with compound 7c exhibiting the most significant reduction, making it a promising candidate for further study. The binding mode of the most promising compound 7c, was determined using the induced fit docking-molecular dynamics (IFD-MD) approach. Regular docking and IFD were also employed for comparison. The IFD-MD analysis revealed that 7c binds to tubulin in a unique binding cavity near that of ABZ, but the benzimidazole ring was fitted much deeper into the binding pocket. Finally, the absolute free energy of perturbation technique was applied to evaluate the 7c binding affinity, further confirming the observed binding mode.

Praziquantel meets Niclosamide: A dual-drug Antiparasitic Cocrystal.

In this paper we report a successful example of combining drugs through cocrystallization. Specifically, the novel solid is formed by two anthelminthic drugs, namely praziquantel (PZQ) and niclosamide (NCM) in a 1:3 molar ratio, and it can be obtained through a sustainable one-step mechanochemical process in the presence of micromolar amounts of methanol. The novel solid phase crystallizes in the monoclinic space group of P21/c, showing one PZQ and three NCM molecules linked through homo- and heteromolecular hydrogen bonds in the asymmetric unit, as also attested by SSNMR and FT-IR results. A plate-like habitus is evident from scanning electron microscopy analysis with a melting point of 202.89 °C, which is intermediate to those of the parent compounds. The supramolecular interactions confer favorable properties to the cocrystal, preventing NCM transformation into the insoluble monohydrate both in the solid state and in aqueous solution. Remarkably, the PZQ - NCM cocrystal exhibits higher anthelmintic activity against in vitro S. mansoni models than corresponding physical mixture of the APIs. Finally, due to in vitro promising results, in vivo preliminary tests on mice were also performed through the administration of minicapsules size M.

In vitro anthelmintic activity of Euphorbia forskalii J. Gay aqueous extracts evaluation on different life stages of Haemonchus contortus.

The use of medicinal plants in the control of gastrointestinal parasitosis is a promising solution for improving the productivity of sheep flocks. In order to evaluate the anthelmintic activity of Euphorbia forskallii, in vitro bioassays were performed on three life stages of Haemonchus contortus. Five aqueous extracts concentrations namely 10 mg/mL; 5 mg/mL; 2.5 mg/mL; 1.25 mg/mL and 0.62 mg/mL were used for adult worm mortality tests. Egg hatch inhibition and L3 larval migration inhibition tests were studied at 5 mg/mL; 2.5 mg/mL; 1.25 mg/mL; 0.62 mg/mL and 0.31 mg/mL. A negative control PBS and a positive control levamisole 2.5 mg/mL were established for each test. A phytochemical screening was performed to determine the presence of some secondary metabolites. The results obtained showed the presence of total polyphenols, total flavonoids and condensed tannins within the aqueous extracts of E. forskalii. A high and significant (P < 0.05) morality rate compared to the negative control with an LC50 of 2.30 mg/mL was obtained. Inhibition of egg hatch and larval migration were high and significant (p < 0.05) compared to the negative control. There was an IC50 of 1.03 mg/mL and 0.92 mg/mL respectively for inhibition of egg hatching and L3 larval migration. The present study revealed the in vitro anthelmintic activity of E. forskalii aqueous extracts and allows us to consider in perspective complementary studies to confirm this activity.

In vitro ovicidal effect of p-coumaric acid from Acacia bilimekii aerial parts against Haemonchus contortus.

Acacia bilimekii is a plant with a high content of protein, fibre, and condensed tannins, making it an excellent feed for small ruminants with anthelmintic potential. This study aimed to evaluate the ovicidal activity of a hydroalcoholic extract (Ab-HA) and fractions from A. bilimekii aerial parts on Haemonchus contortus. The ovicidal activity of the Ab-HA extract and its fractions obtained by chromatographic fractionation were evaluated through the egg hatching inhibition (EHI) test. The results showed that the Ab-HA extract had 91% EHI at 20,000 µg/mL with a mean effective concentration (EC50) of 9260 µg/mL. After liquid-liquid fractionation of Ab-HA extract, the aqueous fraction (Ab-Aq) did not show ovicidal activity, whereas the organic fraction (Ab-EtOAc) showed a better EHI than the Ab-HA extract (98.9% at 2500 µg/mL). Then, the chemical fractionation of Ab-EtOAc allowed obtaining six bioactive fractions (AbR12-17) with an EHI greater than 90% at 1500 µg/mL. The best treatment was AbR15 (98.7% EHI at 750 µg/mL). Chemical analysis by HPLC-PDA of AbR15 indicated the presence of p-coumaric acid and the flavone luteolin as major compounds. Additionally, the commercial p-coumaric acid standard was evaluated in the EHI assay and showed an EHI of 97% at 62.5 µg/mL. Meanwhile, the confocal laser scanning microscopy analysis demonstrated a colocalization effect between p-coumaric acid and the H. contortus embryonated eggs. These results indicate that due to their major chemical compounds (including p-coumaric acid), the aerial parts of the plant A. bilimekii, could be considered as natural potential tool for controlling haemonchosis in small ruminants.

Evaluation of in vivo anthelmintic efficacy of certain indigenous plants against experimentally- induced Ascaridia galli infection in local birds (Gallus domesticus).

The present study was an attempt to evaluate the anthelmintic efficacy of certain indigenous plants against experimentally induced Ascaridia galli infection in local birds (Gallus domesticus). Two indigenous plants viz., Nyctanthes arbor-tristis and Butea monosperma were used based on indigenous technical knowledge (ITK) and ethno-medical uses. Three types of extracts, viz. ethanolic, hydro-ethanolic and aqueous extracts of each of the two plants were prepared and three doses at the rate of 100, 500 and 1000 mg/kg were used for in vivo studies. Piperazine hydrate liquid was used as standard drug at 200 mg/kg body weight. All biochemical and haematological parameters showed a significant increase from 14th day post treatment in birds of all the groups treated with N. arbor-tristis extract at all the doses irrespective of the solvent used. However, the increase in biochemical and haematological parameters with B. monosperma was observed from 21st day of post treatment in all the groups at all the doses. Similarly, piperazine treated group also showed significant increase from 14th day of post treatment. There was significant reduction in egg output in the droppings of the birds treated with ethanolic, hydroethanolic and aqueous extracts of N. arbor-tristis and B. monosperma when compared with infected controls from 7th day onwards.

In vitro evaluation of the effects of methanolic plant extracts on the embryonation rate of Ascaridia galli eggs.

The present study aims to find efficient alternatives to synthetic anthelmintics among ethno-veterinary herbs. Ascaridia galli eggs isolated from the worm uterus were exposed in vitro to methanolic extracts (ME) of nine plant species such as Achillea millefolium (AM), Artemisia absinthium (AA), Artemisia vulgaris (AV), Cicerbita alpina (CA), Cichorium intybus (CI), Inula helenium (IH), Origanum vulgare (OV), Tanacetum vulgare (TV), Tanacetum parthenium (TP). Flubendazole (FL), 0.5% formalin with dimethylsulfoxide and Petri dishes without the addition of reagents were used as positive, negative and untreated control respectively. The effects of the different ME at concentrations 0.500, 0.325, 0.200 mg/ml were assessed on the embryonic development (ED) of the eggs in duplicate. Logit analysis was used to calculate EC50 values. A generalized linear mixed model, having plant species and concentration as fixed effect and day as repeated measure, was used to determine differences in ED. Estimated EC50 was the lowest for FL at 0.11 mg/ml. CA and TV followed with 0.27 mg/ml and 0.32 mg/ml. ED for FL was significantly lower (25%) than that of CA (47%). The analysis showed 0.5 mg/ml of the ME of CA and TV significantly affected the ED at 35% and 42% inhibitions respectively. The ED for all ME showed similar pattern i.e., relatively higher efficacy in the first experimental week compared to the rest of the experimental period. The effect from all multicomponent extracts is time and dose dependent. The plants have promising results in inhibiting ED, contributing to the identification of alternative anthelmintic treatments.

In vivo anthelmintic potentials of Gongronema latifolium and Picralima nitida against gastrointestinal parasite (Heligmosomoides bakeri).

Gastrointestinal parasites which are responsible for soil-transmitted helminthiases cause illness globally. The control of helminthiasis depends on mass distribution of anthelmintics which has been reported for its resistance, toxicity and low efficacy. In this study, anthelmintic potentials of Gongronema latifolium and Picralima nitida which have wide application in traditional medicine were determined in vivo using Heligmosomoides bakeri a naturally occurring gastro-intestinal parasite of rodents that is closely related to highly prevalent human nematode parasites. Extracts of P. nitida at 500 mg/kg had higher (p< 0.05) chemosuppression (92.45 %) than extracts of G. latifolium (65.82 %) and was highly comparable to albendazole (92.61 %). As the dose of the extracts increased from 300 to 500 mg/kg body weight, chemosuppression of 84.91 % and 92.45 % (P. nitida) and 43.54 % and 65.82 % (G. latifolium) respectively were produced. The extract of P. nitida gave deparasitization rates (p<0.05) of 72.60 % and 77.16 % at 300g/kg and 500mg/kg of body weight respectively. The glucose level and protein content reduced (p<0.05) in mice treated with extract of P. nitida when compared with extract of G. latifolium and untreated mice. Phytochemical screening revealed that P. nitida and G. latifolium contained flavonoids, alkaloids, saponins, tannins and polyphenols. Acute toxicity studies showed that Gongronema latifolium and Picralima nitida have no apparent toxic effect in mice even at the dose of 5000 mg/kg. Extracts of P. nitida and G. latifolium have anthelmintic properties that are dose-dependent, and this could offer potential lead for the development of safe, effective and affordable anthelmintics.

Bioassay-Guided Isolation of Anthelmintic Components from Semen pharbitidis, and the Mechanism of Action of Pharbitin.

Parasitic helminths continue to pose problems in human and veterinary medicine, as well as in agriculture. Semen pharbitidis, the seeds of Pharbitis nil (Linn.) Choisy (Convolvulaceae), is a well-known traditional Chinese medicinal botanical preparation widely used for treating intestinal parasites in China owing to its desirable efficacy. However, the anthelmintic compounds in Semen pharbitidis and their mechanism of action have not been investigated yet. This study aimed to identify the compounds active against helminths from Semen pharbitidis, and to establish the mechanism of action of these active compounds. Bioassay-guided fractionation was used to identify the anthelmintic compounds from Semen pharbitidis. The anthelmintic assay was performed by monitoring Caenorhabditis elegans (C. elegans) motility with a WMicrotracker instrument. Active compounds were identified by high-resolution mass spectrometry. Several (analogues of) fragments of the anthelmintic compounds were purchased and tested to explore the structure-activity relationship, and to find more potent compounds. A panel of C. elegans mutant strains resistant to major currently used anthelmintic drugs was used to explore the mechanism of action of the active compounds. The bioassay-guided isolation from an ethanol extract of Semen pharbitidis led to a group of glycosides, namely pharbitin (IC50: 41.0 ± 9.4 µg/mL). Hit expansion for pharbitin fragments yielded two potent analogues: 2-bromohexadecanoic acid (IC50: 1.6 ± 0.7 µM) and myristoleic acid (IC50: 35.2 ± 7.6 µM). One drug-resistant mutant ZZ37 unc-63 (x37) demonstrated a ~17-fold increased resistance to pharbitin compared with wild-type worms. Collectively, we provide further experimental scientific evidence to support the traditional use of Semen pharbitidis for the treatment of intestinal parasites. The anthelmintic activity of Semen pharbitidis is due to pharbitin, whose target could be UNC-63 in C. elegans.

Anthelmintic Activity, Cytotoxicity, and Phytochemical Screening of Plants Used to Treat Digestive Parasitosis of Small Ruminants in Benin (West Africa).

Medicinal plants continue to be used alone or in combination with veterinary drugs to treat animal ailments, especially in developing countries where livestock farmers often lack access to modern veterinary services and drugs. In addition, digestive parasitosis remain a major constraint for small ruminant livestock. The objective of this study was to screen the anthelmintic activity of the main plants used in the treatment of the digestive parasitosis of small ruminants in Benin. A total of 40 extracts were prepared using the successive maceration of 10 plants in four solvents of increasing polarity. The phytochemical screening of the plants was performed, and the anthelmintic activity of the extracts was evaluated on L3 larvae of Haemonchus contortus. The cytotoxicity of the 40 extracts was determined on WI38 noncancerous fibroblast cells using the MTT assay, and the total phenol content (TPC), total flavonoid content (TFC), and condensed tannin content (CTC) were quantified in the most effective extracts using colorimetric methods. The results show that the plants contained tannins, flavonoids, and triterpenoids which may, in part, justify their anthelmintic activities. All plants gave active extracts at the highest concentration tested (1200 µg/mL). Methanol (MeOH) extracts were, in general, more effective than the hexane (HEX), dichloromethane (DCM), and aqueous (H2O) ones in inhibiting larval migration, with the MeOH extracts of Terminalia leiocarpa, Adansonia digitata, and Momordica charantia being the most effective. Nevertheless, the MeOH extract of M. charantia was highly cytotoxic at the concentration of 100 µg/mL. The anthelmintic activity of M. charantia, Vitex doniana, and Caesalpinia bonduc was studied on H. contortus for the first time. These results provide scientific information that can be used for better valorization of the anthelmintic potential of the studied plants and to initiate the process of the identification of new anthelmintic molecules.

In vitro and in vivo anthelmintic properties of Caesalpinia coriaria fruits against Haemonchus contortus.

The purpose of the present study was to assay the in vitro and in vivo anthelmintic activity (AA) of Caesalpinia coriaria (Cc) mature fruits against the nematode Haemonchus contortus (Hc). The Hc infective larvae were used to assess the in vitro AA through larval mortality assay. The exposure of larvae to the different treatments was performed in 96-well microtitration plates. The treatments were as follows: hydroalcoholic extract (HA-E, at 25-100 mg/mL), aqueous fraction (Aq-F, at 12.5-50 mg/mL), organic fraction (EtOAc-F at 12.5-50 mg/mL), compounds (1, methyl gallate and 2, gallic acid at 1.25-10 mg/mL), positive control (ivermectin at 5 mg/mL) and two negative controls (distilled water and 4% methanol). After exposure, dead and live larvae were quantified and results were compared to their controls. The in vivo assay was carried out by a faecal egg count reduction test (FECRT); artificially infected goat kids (F1: Boer x Nubia) were treated with Cc ground dried fruits to assess the AA. The treatments were established as follows: G1-untreated goats (negative control), G2-goats dewormed with ivermectin (positive control), G3-goats fed with Cc mature fruits (10% of their diet). Results in both in vitro and in vivo assays were analysed using an ANOVA through random design, applying a general linear model and mixed models. The in vitro results showed an evident larvicidal effect of the HA-E, EtOAc-F from Cc, indicating that the compound responsible for the AA was gallic acid. The results of the in vivo study corroborated the anthelmintic properties of Cc, reaching 78.6% reduction in the elimination of Hc eggs per gram of faeces. This plant represents a potential natural anthelmintic for the control of haemonchosis in goats under grazing conditions. Future studies should standardise the Cc extract or dried fruits for use in the management of nematodiasis in goat herds.

Phenolic Acids and Flavonoids from Pithecellobium dulce (Robx.) Benth Leaves Exhibit Ovicidal Activity against Haemonchus contortus.

Pithecellobium dulce (Robx.) Benth is an arboreal legume used in traditional medicine for the treatment of several ailments, including a number of intestinal disorders, and as a natural deworming. The objective of this study was to evaluate the ovicidal activity of a hydroalcoholic extract (HA-E) and its fractions (aqueous, Aq-F and organic, EtOAc-F) from P. dulce leaves, as well as subfractions (C1F1-C17) obtained from EtOAc-F against Haemonchus contortus eggs. The HA-E, Aq-F, and EtOAc-F were assessed at 0.03-5.00 mg/mL and subfractions (C1F1-C17) were assessed at 0.06-1.00 mg/mL. The HA-E and Aq-F showed an ovicidal activity close to 100% at 2.5 mg/mL, and EtOAc-F displayed the highest anthelmintic effect (100% at 0.25 mg/mL). Meanwhile, the sub-fractions with the highest ovicidal effect were C1F6, C1F9, C1F10, C1F11, and CIF13. The main compounds identified in the most active fractions (C1F9, C1F11, and C1F13) were kaempferol (1), quercetin (2), coumaric acid (3), ferulic acid (4), luteolin 7-O-rhamnoside (5), quercetin 3-O-rhamnoside (6), and a caffeoyl derivate (NI). The results indicate that P. dulce leaves exhibit a potent anthelmintic activity and contain bioactive compounds able to inhibit egg hatching in H. contortus. Therefore, this plant could be used for the control of gastrointestinal nematodes in small ruminants.

In Vitro Anthelmintic Activity of a Hydroalcoholic Extract from Guazuma ulmifolia Leaves against Haemonchus contortus.

The purpose of the present study was to assess the ovicidal and larvicidal activity of a hydroalcoholic extract (HAE) and their fractions (aqueous, Aq-F and organic, EtOAc-F) from Guazuma ulmifolia leaves using Haemonchus contortus as a biological model. The egg hatching inhibition (EHI) and larval mortality against infective larvae (L3) tests were used to determine the anthelmintic effect of the treatments. The extract and fractions were tested at different concentrations against eggs and L3. Additionally, distilled water and methanol were used as negative controls and ivermectin as a positive control. The extract and fractions were subjected to HPLC analysis to identify the major compounds. The HAE displayed the highest ovicidal activity (100% EHI at 10 mg/mL). Fractionation of the HA extract allowed increasing the nematicidal effect in the EtOAc-F (100% EHI at 0.62 mg/mL and 85.35% mortality at 25 mg/mL). The phytochemical analysis of the extract and fractions revealed the presence of kaempferol, ethyl ferulate, ethyl coumarate, flavonol, luteolin, ferulic acid, luteolin rhamnoside, apigenin rutinoside, coumaric acid derivative, luteolin glucoside, and quercetin glucoside. These results suggest that G. ulmifolia leaves could be potential candidates for the control of H. contortus or other gastrointestinal parasitic nematodes.

Potential effects and relevant lead compounds of Vigna mungo (L.) Hepper seeds against bacterial infection, helminthiasis, thrombosis and neuropharmacological disorders.

Multidrug-resistant bacterial infections, helminthiasis, thrombosis, anxiety and insomnia are some of the major global health concerns. Vigna mungo (L.) Hepper (VM) has been used traditionally to treat microbial infection, helminthic disorder, schizophrenia, memory loss, and blood circulatory problem. This research aims to discover antibacterial, anthelmintic, thrombolytic and neuropharmacological effects of the methanol extract of Vigna mungo seeds (MESVM), and also in-silico prediction of relevant lead compounds by molecular docking and ADME/T analysis. The crude extracts and subsequent fractions of MESVM were investigated for antibacterial activity by disc diffusion method, anthelmintic activity by paralysis and death test on earthworms, and thrombolytic activity by in vitro blood clot dissolution test. Open-field test and elevated plus maze test were performed for evaluating anxiolytic activity of the extracts. Using molecular docking, ligand poses of selected VM seeds' phytoconstituents were predicted targeting tubulin, GlcN-6-P synthase, and human tissue plasminogen activator proteins for anthelmintic, antibacterial, and thrombolytic activity, respectively. In the antibacterial activity test, the MESVM at 10000 µg/mL concentration created highest and significant (P < 0.001) zone of inhibition against Staphylococcus aureus (15.42 mm) and Escherichia coli (12 mm) compared with tetracycline. The MESVM exhibited remarkable anthelmintic activity at 50 mg/mL concentration with 35.4 min paralysis time, 75.2 min death time and were closer to the durations of standard drug albendazole. No test extract showed anxiolytic activity. In thrombolytic activity test, all concentrations of MESVM produced clot lytic activity with high significance (P < 0.001) in comparison with the blank. In docking, 2'-hydroxygenistein, cyclokievitone hydrate, and aureol displayed maximum affinity to the target proteins for anthelmintic, antibacterial, and thrombolytic activity, respectively. This research revealed that the MESVM demonstrated potential anthelmintic, antibacterial and thrombolytic effects that confirmed the folkloric uses of VM and the found relevant lead compounds might be further optimized in future drug development.

In Vitro Anthelmintic Activity of Sea Buckthorn (Hippophae rhamnoides) Berry Juice against Gastrointestinal Nematodes of Small Ruminants

Gastrointestinal nematodes are one of the major threats in small ruminant breeding. Their control is difficult due to the development of anthelmintic resistance, and the search for new molecules endowed with anthelmintic activity (AH) is considered a priority. In this context, we evaluated the in vitro AH activity of two commercial sea buckthorn (Hippophae rhamnoides) berry juices, namely SBT and SBF. The in vitro evaluation was based on the egg-hatch test and larval exsheathment assay at different concentrations. Data were statistically analysed, and the EC50 was calculated. Chemical analyses were performed to evaluate the total polyphenol content of the juices and chemical profile of the most represented compounds. The role of the polyphenolic fraction in the anthelmintic activity of the juices was also assessed. At the highest concentrations, the activity of SBT was high in both tests and comparable to that observed in the thiabendazole-treated positive controls, while SBF showed a lower efficacy. Glycosylated isorhamnetin and quercetin were the most represented polyphenolic compounds in both juices. In conclusion, both H. rhamnoides berry juices tested in this study showed interesting anthelmintic properties in vitro.

Ultrastructural tegumental changes of Trichinella spiralis adult and larval stages after in vitro exposure to Allium sativum.

Albendazole (ALB) is the most used therapeutic drug for trichinellosis treatment, but it has many drawbacks. Hence, the search for alternative natural compounds is a goal for researchers. The present work aimed to test the in vitro anthelmintic effect of Tomex (Allium sativum) against adult and muscular larva of Trichinella spiralis (T. spiralis). We incubated adult forms and muscular larvae of T. spiralis with Tomex at concentrations 10, 50, and 100 µg/mL to evaluate the changes that happened on the cuticle using a scanning electron microscopy (SEM). Although ALB was very effective, Tomex also affected motility and the tegumental structure of T. spiralis, which was in proportion to its concentration and incubation time, as Tomex started the mortality of muscular larvae and adult stages after 4 h at 50 µg/mL, and after 6 h at10 µg/mL. A 50% mortality rate of muscular larvae was recorded after 6 h at 100 µg/mL. However, a 50% mortality rate was recorded after 12 h at 10 µg/mL for the adult stage. Adult worms and muscular larvae of T. spiralis incubated with 100 µg/mL of Tomex displayed loss of normal annulations and creases of the cuticle, multiple vesicles, widening of longitudinal furrow space, and multiple minor loss blebs. Our results suggested that Tomex can be a therapeutic agent against adults and larvae stages of T. spiralis.