New therapeutic approaches are necessary to control strongyloidiasis due to the side effects of, and resistance to, currently available drugs thiabendazole, albendazole, and ivermectin. This study examined the anthelmintic properties of extracts and isolated compounds from Siparuna guianensis against Strongyloides venezuelensis eggs and larvae, using the egg hatching test (EHT) and larval motility test (LMT). Albendazole (0.025 mg/ml) and ivermectin (0.316 mg/ml) were used as the positive controls for the EHT and LMT assays, respectively. Strongyloides venezuelensis eggs or larvae (±50 specimens) were treated with ethanol extract (0.05–1.0 mg/ml), ethyl acetate and aqueous fractions (0.05–0.8 mg/ml), essential oil (0.2–1.0 mg/ml) and α-bisabolol (0.2–1.0 mg/ml) from S. guianensis, and analysed by optical microscopy after 48 h (EHT), or after 24, 48 and 72 h (LMT). All the tested compounds exhibited ovicidal activity equivalent to the positive control and changed the morphology of the eggs. The S. guianensis ethanol extract and aqueous fraction were as effective as the positive control. Phytochemical analysis of the ethanol extract and fractions revealed the presence of phenolic compounds, tannins and flavonoids. Therefore, S. guianensis is effective against S. venezuelensis eggs and larvae in vitro, and can be considered as a potential alternative treatment for strongyloidiasis.

The toxicity of water–ethanol extracts of garlic (Allium sativum), ginger (Zingiber officinale), basil (Ocimum basilicum), bitter chaparro (Castela tortuousa), onion (Allium cepa) and papaya (Carica papaya) against adults, eggs and oncomiracidia of Neobenedenia spp. parasites was examined. Parasites were exposed to continuous immersion and treated as follows: extracts were tested at three dilutions: 1:10, 1:50 and 1:100 made with filtered seawater (35 g l–1); ethanol (70%) was evaluated at the same dilutions of 1:10 (7% ethanol), 1:50 (1.4% ethanol) and 1:100 (0.07% ethanol) and a seawater (35 g l–1) control. The antiparasitic effect was measured on: (1) adult survival, egg production and time to detachment from the culture vessel; (2) egg development and cumulative egg hatching; and (3) oncomiracidia survival. All three dilutions of ginger and dilutions 1:100 and 1:50 of basil extract reduced adult survival in vitro, time to detachment from the surface of the culture vessel, egg production and oncomiracidia survival. Bitter chaparro extract reduced adult egg production and oncomiracidia survival. Hatching success was significantly reduced (P < 0.05) in basil extract (1:100) to 86.6% compared to the seawater control (100%). Dilutions 1:10 of ginger and basil exhibited the highest impact on the biological parameters of Neobenedenia sp. Our study demonstrates that water–ethanol extracts of ginger, basil and bitter chaparro are toxic against Neobenedenia sp. life stages.

This study assessed the in vitro anthelmintic (AH) activity of methanol and acetone:water leaf extracts from Annona squamosa, A. muricata and A. reticulata against Haemonchus contortus eggs. The egg hatch test was used to determine the effective concentrations required to inhibit 50% of eggs hatching (EC50). The role of polyphenols on AH activity was measured through bioassays with and without polyvinylpolypyrrolidone (PVPP). Methanolic extracts mainly caused the death of eggs at the morula stage (ovicidal activity). Meanwhile, acetone:water extracts caused egg-hatching failure of developed larvae (larvae failing eclosion (LFE) activity). The lowest EC50 values against H. contortus eggs were observed for the methanolic extracts from A. reticulata and A. muricata (274.2 and 382.9 µg/ml, respectively). From the six extracts evaluated, the methanolic extracts of A. muricata, A. reticulata and A. squamosa showed the highest ovicidal activity, resulting in 98.9%, 92.8% and 95.1% egg mortality, respectively. When the methanolic extract of A. squamosa was incubated with PVPP, its AH activity increased. Similarly, when acetone:water extracts of A. muriata and A. reticulata were incubated with PVPP, their LFE activity increased. Alkaloids were only evident in methanolic extracts, irrespective of PVPP incubation. The presence of acetogenins was not observed. In conclusion, methanolic extracts obtained from leaves of A. muricata, A. reticulata and A. squamosa showed ovicidal activity affecting the morula of H. contortus eggs, with minor LFE activity. Meanwhile, acetone:water extracts showed mostly LFE activity, with a lower proportion of ovicidal activity.

The chemical treatment of gastrointestinal parasitic diseases has been undermined by increasing resistance and high toxicity. There is an urgent need to search for alternative natural sources for the treatment of such parasites. In this respect, the present study aims to quantify phenolic compounds of chamomile (Matricaria recutita L.) and to study their in vitro anti-oxidant and anthelmintic activities in solvents with increasing polarity. In vitro determination of anti-oxidant capacity was carried out using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2-azino-bis-(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) radical cation methods. In vitro anthelmintic activity was investigated on egg-hatching inhibition and loss of motility of adult worms of Haemonchus contortus from sheep. The results showed that methanolic and aqueous extracts contain more total polyphenols, total flavonoids and condensed tannins than chloroformic and hexanic extracts. ABTS and DPPH assays showed that methanolic extracts had the highest anti-oxidant potency (IC50 = 1.19 μg/ml and 1.18 μg/ml, respectively). In vitro anthelmintic activity showed that both methanolic (IC50 = 1.559 mg/ml) and aqueous (IC50 = 2.559 mg/ml) extracts had the greatest effect on egg hatching and motility of worms (100% after 8 h post exposure at 8 mg/ml). A significant and positive correlation between DPPH and ABTS tests was observed for all tested extracts. Therefore, total phenolic, total flavonoid and condensed tannin values were correlated with IC50 from both ABTS and DPPH, and with inhibition of egg hatching. To our knowledge, this report is the first of its kind to deal with in vitro anthelmintic activities of chamomile extracts.

Hymenolepis nana is a common intestinal tapeworm that affects humans. Drugs are available for the treatment of this infection, including praziquantel (PZQ), nitazoxanide and niclosamide. Although the drug of choice is praziquantel, due to its high cure rates, indicators of the development of PZQ resistance by different parasites have begun to appear over recent decades. Therefore, this study was a trial to find an alternative to PZQ by assessing the activity of the crude aqueous extract of the medicinal herb Artemisia absinthium against H. nana. In vitro, the extract was used against adult worms at concentrations of 1 and 5 mg/ml, in comparison with 1 mg/ml of PZQ. The times of worm paralysis and death were determined. Ultrastructural morphological changes were studied using transmission electron microscopy (TEM). For the in vivo study, infected mice were divided into untreated, PZQ-treated and A. absinthium-treated groups (400 mg/kg and 800 mg/kg). Pre- and post-treatment egg counts per gram of faeces (EPG) were performed; then, the reduction percentages of the EPG and worm burden were calculated. The best results were obtained with praziquantel. Artemisia absinthium induced worm paralysis, death and ultrastructural alterations, such as tegumental damage, lipid accumulation, and destruction of the nephridial canal and the intrauterine eggs, in a dose-dependent manner. Additionally, significant reductions in the EPG and worm burden were recorded in A. absinthium-treated mice. Although the results obtained with A. absinthium were promising and comparable to PZQ, further studies using different extracts, active ingredients and concentrations against different parasites should be conducted.

The in vitro larvicidal and in vivo anthelmintic effects of Oxalis tetraphylla hydroalcoholic extract (HE), against Haemonchus contortus in experimentally infected lambs, were assessed. We used a microtitration plate method, comprising the following two stages. Stage 1: 20 μl of water containing 200 sheathed H. contortus infective larvae (ShHcl) were deposited in every well of three series; then, the series 2 and 3 wells were treated with 80 μl 1% ivermectin and O. tetraphylla HE at 20 mg/ml, respectively. Stage 2: the same procedure was performed replacing the ShHcl with exsheathed larvae (ExShHcl). Evaluations were performed after 24 and 48 h. The total numbers of dead and live larvae were counted. A second experiment evaluated the reduction in nematode egg populations in the faeces of lambs treated orally with the O. tetraphylla HE. The 27 lambs used were divided into Groups 1, 2 and 3 (n = 9), which were administered water (positive control), levamisole 1 m (7.5 mg/kg body weight (BW), as a unique dose) and O. tetraphylla HE (20 mg/kg BW), respectively. The plant HE was administered daily for 8 days. The in vitro assay showed 80.9% and 86.5% larval mortality of ShHcl after 24 and 48 h, respectively, while the corresponding mortality values for ExShHcl were 97 and 99%, respectively. The in vivo assay showed variability in the eggs/gram of faeces (epg) values; however, at the end of the trial, the average reduction in the epg values of the O. tetraphylla HE group was 45.6% (P < 0.05). Oxalis tetraphylla HE contains compounds that belong to the flavonol group with anthelmintic activity.

Schistosomiasis is a chronic debilitating parasitic disease that causes hepatic damage and is known to be endemic in developing countries. Recent control strategies for schistosomiasis depend exclusively on chemotherapeutic agents, specifically praziquantel. Unfortunately, praziquantel has low efficacy in the early phase of infection, and resistance to treatment is increasingly reported. The aim of this work was to find an alternative treatment by assessing the in vivo activity of aqueous extract of Callistemon citrinus against Schistosoma mansoni in both prepatent and patent phases in experimentally infected mice. The study was conducted on 80 male BALB/c albino mice divided into eight groups. Callistemon was administered at a dose of 200 mg/kg on days 14 and 45 post infection as a single therapy and in combination with praziquantel. Porto-mesenteric worm burden, hepatic and intestinal egg counts, hepatic granuloma number and diameter, and oogram pattern were assessed to evaluate the anti-schistosomal properties of C. citrinus. Liver enzymes and total bilirubin were tested to assess hepatoprotective effects. Results revealed that the use of C. citrinus was associated with a significant decrease in worm burden and tissue egg load together with an increased percentage of dead eggs. In addition, there was a significant reduction in granuloma formation. Callistemon also led to a significant improvement in liver function. The best results were obtained when C. citrinus was given in the prepatent phase of infection and when combined with praziquantel. Although the effects of C. citrinus are considered to be promising, further studies using different extracts, active ingredients and doses are needed.

The in vitro nematicidal effect of Chenopodium ambrosioides and Castela tortuosa n-hexane extracts (E-Cham and E-Cato, respectively) on Haemonchus contortus infective larvae (L3) and the anthelmintic effect of these extracts against the pre-adult stage of the parasite in gerbils were evaluated using both individual and combined extracts. The in vitro confrontation between larvae and extracts was performed in 24-well micro-titration plates. The results were considered 24 and 72 h post confrontation. The in vivo nematicidal effect was examined using gerbils as a study model. The extracts from the two assessed plants were obtained through maceration using n-hexane as an organic agent. Gerbils artificially infected with H. contortus L3 were treated intraperitoneally with the corresponding extract either individually or in combination. The results showed that the highest individual lethal in vitro effect (96.3%) was obtained with the E-Cham extract at 72 h post confrontation at 40 mg/ml, followed by E-Cato (78.9%) at 20 mg/ml after 72 h. The highest combined effect (98.7%) was obtained after 72 h at 40 mg/ml. The in vivo assay showed that the individual administration of the E-Cato and E-Cham extracts reduced the parasitic burden in gerbils by 27.1% and 45.8%, respectively. Furthermore, the anthelmintic efficacy increased to 57.3% when both extracts were administered in combination. The results of the present study show an important combined nematicidal effect of the two plant extracts assessed against L3 in gerbils.

Infections by gastrointestinal nematodes negatively affect small ruminant health and at the same time cause substantial economic losses worldwide. Because resistance to conventional anthelmintic compounds is growing, target studies evaluating the effectiveness of alternative ingredients of botanical origin on gastrointestinal nematodes are needed. In this study, we evaluated the in vitro anthelmintic activity of Ruta chalepensis L. extracts on the third-stage larvae of sheep gastrointestinal nematodes. A methanol extract showed the highest anthelmintic activity, with an EC50 = 0.10 ± 0.06 mg/ml after 96 h, while the essential oil had an EC50 = 1.45 ± 1.22 mg/ml after 48 h. Moreover, three secondary metabolites of the essential oil, i.e. 2-decanone, 2-nonanone and 2-undecanone, showed EC50 values of 0.07 ± 0.06, 0.25 ± 0.29 and 0.88 ± 0.73 mg/ml at 24 h, respectively. The present study indicated that the R. chalepensis methanol extract, the essential oil and its metabolites 2-decanone, 2-nonanone and 2-undecanone showed promising anthelmintic activity on gastrointestinal nematodes.

This study assessed the efficacy of closantel vis-à-vis herbal extracts with known anti-parasitic properties, against fenbendazole-resistant nematodes in goats maintained under a semi-intensive system of management at the University goat farm, Jabalpur. Fifty goats were randomly assigned to five groups, each comprising 10 animals, irrespective of their breed, age and sex. Each animal in Group I, II and III was orally administered with aqueous leaf extracts of neem (Azadirachta indica) at 1 g/kg body weight, sitaphal (Annona squamosa) at 1.5 g/kg body weight and tobacco (Nicotiana tabacum) at 1 g/kg body weight, respectively, whereas Group IV was an untreated control group. Each animal in Group V was orally treated with closantel at 10 mg/kg body weight. During the course of the study, all animals were maintained under an identical semi-intensive system of management. Compared to the untreated control group (Group IV), there was no conspicuous reduction in post-treatment (day 10) faecal egg counts (FEC) in animals administered with the herbal extracts (Groups I, II and III), which is suggestive of poor anti-parasitic activity. However, using the faecal egg count reduction test (FECRT), the overall efficacy of closantel was recorded as 95.64%. This supports the rotational use of closantel as a preferred choice over the benzimidazole group of anthelmintics and/or herbal extracts to meet the acute challenge of in situ development of drug-resistant gastrointestinal nematodes, especially Haemonchus contortus.

In this study, we evaluated the in vivo comparative efficacy of ivermectin and Nigella sativa extract against helminths in Aseel chickens, and the effects of helminths on blood parameters before and after treatment in Aseel chickens. Forty naturally infected adult Aseel chickens were randomly divided into four groups (n = 10 each): group A (ivermectin at 300 μg/kg); group B (N. sativa extract at 200 mg/kg); group C (ivermectin at 300 μg/kg + N. sativa extract at 200 mg/kg); group D was kept as a positive control to monitor time-related changes. On day 28 post treatment, the mean percentages of faecal egg-count reduction (FECR %) in groups A, B and C were recorded as 93.58, 88.09 and 100.00%, respectively. Further data analysis showed significantly higher efficacy in group C (100 ± 0.00%) than in groups A and B (P < 0.001). Highly significant (P < 0.001) improvements in mean percentage values of packed cell volume (PCV %) were recorded in groups A and C on days 14 and 28 post treatment. Meanwhile, the improvements in mean values of haemoglobin (Hb) concentration in groups A, B and C were highly significant (P < 0.001) when compared to that of group D on day 28 post treatment. The synergistic combination of ivermectin and N. sativa extract possessed greater efficacy than either ivermectin or N. sativa extract used alone. Furthermore, both PCV % and Hb concentration values gradually increased in the treated groups compared to the control group, in which PCV % and Hb concentration gradually decreased throughout the trial.

This study describes the effects of extracts and fractions of Persea willdenovii leaves against goat gastrointestinal nematodes and their cytotoxicity on Vero cells. The in vitro ovicidal and larvicidal activities of the crude ethanolic, hexane, ethyl acetate (EAE), butanolic and residual hydroethanolic extracts were assessed through the inhibition of egg hatching and larval motility assays. The most active extract (EAE) was then fractionated by chromatography in an open column containing silica gel, to furnish six fractions (Fr1–Fr6), which were also tested. The cytotoxicity of active extracts and fractions was determined using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and trypan blue exclusion assay. The EAE and two fractions (Fr1 and Fr2) showed inhibitory activity in the egg hatching of gastrointestinal nematodes of goats in a concentration-dependent manner. The effective concentrations for 50% inhibition (EC50) of egg hatching were 2.3, 0.12 and 2.94 mg/ml for EAE, Fr1 and Fr2, respectively. All extracts and fractions were not effective in inhibiting 50% of motility of infective larvae. EAE and Fr2 had IC50 values (50% inhibitory concentration) of 4.95 and 2.66 mg/ml, respectively. Fr1 showed a slight cytotoxic effect (cellular inviability <30%) only after 48 h of treatment (MTT test). Gas chromatography–mass spectrometry (GC–MS) analysis showed the presence of six fatty acid ethyl esters, a fatty acid methyl ester and a long-chain ketone in the most active fraction. These constituents identified in P. willdenovii can be related to the high ovicidal activity and relatively non-toxic effect of the extracts.