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Effect of different concentrations of acetone:water extracts of Acacia collinsii (a), A. pennatula (b), Bunchosia swartziana (c), Gymnopodium floribundum (d), Havardia albicans (e), Leucaena leucocephala (f), Lysiloma latisiliquum (g), Mimosa bahamensis (h), Piscidia piscipula (i), and Senegalia gaumeri (j) on the proportion of Haemonchus contortus eggs hatched in vitro. Each point represents the mean (± SD) of six replicates per extract concentration
Source publication
Nutraceutical plants provide nutrients for the animal as well as secondary compounds that can affect the biology and survival of gastrointestinal nematodes (GIN). Current screening of nutraceutical plants is based on in vitro evidence of anthelmintic (AH) activity against different life stages of GIN, but nutritional information is omitted or scarc...
Citations
... Considering that one of the infected groups received PEG in their diet but still reduced their GIN egg excretion, Torres-Fajardo et al. (2018) hypothesized that other PSM other than CT, which are not neutralized by PEG, may also possess anthelmintic activity against H. contortus. Recent studies confirmed that PSM other than CT display AH activity against H. contortus eggs and L 3 larvae (Castañeda-Ramírez et al., 2018. These outcomes reinforced the complexity and multifactorial nature of the goat's feeding behaviour as well as the existence of unknown interactions between dietary compounds themselves and between PSM and the GIN population. ...
... Arboreal legumes represent an important source of nutrients such as protein and fibre for ruminants; because of the contents of secondary compounds, such as polyphenols, flavonoids and saponins, they are a nutraceutical feed for livestock (Castañeda-Ramírez et al., 2018). In this context, there are different reports on the in vitro and in vivo effect of several legumes with a high anthelmintic effect (Brito et al., 2018;Castillo-Mitre et al., 2021). ...
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.
... This study evaluated the nutritional characteristics of ruminant feeds such as CP, ash, NDF, ADF, CT, TT, and TP ( [27]. The evaluation of D. eriantha hay (Di) showed a low CP content (T2 = 5.86%) but the latter was expected as most tropical grasses show low CP content (< 5% CP) [28]. ...
... The effect against eggs observed with these extracts was associated with larvae trapped inside the eggs, which is the most common effect reported for hydroalcoholic extracts of different tropical plants when confronted to H. contortus [2]. It is also important to mention that the few larvae that were able to hatch from eggs showed structural damage (Fig. 1), which is consistent with previous reports obtained with leaf extracts from tropical trees [27,34], and also with extracts obtained from Theobroma cacao leaves and husks [10]. ...
Purpose
This study evaluated the in vitro anthelmintic (AH) activity against Haemonchus contortus of ten extracts obtained from coffee pulp waste (Coffea canephora (Co)), maize comb waste (Zea mays (Zm)), pangola grass hay (Digitaria eriantha Steud (Di) and different mixtures of those materials.
Methods
Three batches prepared with individual feedstuffs (T1, T2 and T3), 3 batches formed with 2 feedstuffs (50:50 proportion; T4, T5 and T6), a batch combining 3 feedstuffs (T7) and 3 batches combining 3 feedstuffs (T8, T9 and T10). The batches of individual feedstuffs and mixtures were used to determine their chemical composition as well as preparing 10 methanol–water (70–30%) extracts. The in vitro tests used against H. contortus were egg hatch test (EHT), larval mortality test (LMT) and larval exsheathment inhibition test (LEIT).
Results
Chemical composition suggested that the nutritional value of Co and the batches including Co (T1, T4, T6 to T10) could be used for ruminant nutrition, but the Di and Zm showed very poor nutritional potential unless they are combined with Co. Extracts showing activity against eggs were T4 and T8 (P < 0.05). Significant L3 mortality was reported for extracts T1, T3, T4 and T8 (P < 0.05). Extracts of T3, T4, T7, T8 and T10 showed an EC50 < 1000 μg/mL for the L3 exsheathment inhibition (P < 0.05). Chemical analyses showed the presence of coumarins and flavonoids in all the extracts.
Conclusion
Extracts obtained from T4 and T8 showed the best overall activity in the three in vitro tests against H. contortus and a good nutritional quality that could be suited for ruminant nutrition.
Graphical Abstract
... The values of CT content in the present study were within the range reported previously for the TDF of Yucatán [6,10,34]. The RH maximum recorded at the September harvest corresponded with the CT content, which was highest in September, regardless of leaf age ( Table 1). ...
In the present study, the effects of macro- and microclimatic conditions, month of harvest, and leaf age at harvest on the bromatological composition and polyphenol content of Gymnopodium floribundum leaves were evaluated. Leaves were harvested in December 2017 and 2018 and March, June, and September 2018. At each harvest, three composite samples of mixed-age leaves were collected from 12 trees (four trees for each sample), and the sampling was repeated on day 90 post- harvest to collect 90-day-old leaves. Fresh and dry matter, crude protein, acid and neutral detergent fibers (ADF and NDF, respectively), lignin, total tannins, condensed tannins (CT), total phenols, in vitro dry matter (IVDMD) and organic matter (IVOMD) digestibility, and metabolizable energy (ME) were estimated. Rainfall, relative humidity, and microhumidity were associated with chemical composition. IVDMD, IVOMD, and ME were highest in leaves sampled in March regardless of age (p < 0.001). Water content, ADF, NDF, and lignin were highest in the leaves sampled in September, regardless of age (p < 0.05), suggesting that leaves require more structural support in the rainy season. CT content was highest in September in the mixed-age leaves and in September and December in the 90-day-old leaves (p < 0.05). A high fiber and CT content during the period of rapid leaf growth could deter herbivory.
... Extracts from various parts of L. leucocephala had therapeutic effects on parasitic nematodes found in sheep [18,19]. In addition to nematodes [20][21][22], there are also articles related to protozoa [23], but, unfortunately, no clear curative effect has been found against nematodes and cestodes. In addition to worms, components of L. leucocephala were also found to be effective against some insects of medical importance [24]. ...
Background/purpose:
Hymenolepis nana is globally distributed. Leucaena leucocephala has been studied as a treatment, including the nematodes and protozoa, but no research results are related to cestodes. Therefore, the aim of this study was to target H. nana.
Methods:
The natural components of L. leucocephala were isolated and added to H. nana, which was cultured in vitro, to observe changes in the mortality, motility, and morphology. BALB/c male mice infected with H. nana were treated with effective components of L. leucocephala for 10 days, and the changes were recorded. After the mice were sacrificed, the spleen weight was measured, and a primary culture was performed for the subsequent cytokine and chemokine testing.
Results:
The experiment found that 132-hydroxy-(132-S)-pheophytin a and aristophyll-C have clear cestocidal effects in vitro. 132-hydroxy-(132-S)-pheophytin a has been shown to be effective at reducing parasite populations and eliciting host immune responses in vivo. IL-2, IL-4, IL-5, IL-6, IL-10, IL-13, IL-17, MCP-1, IFN-γ, TNF-α, MIP-1α, and GM-CSF in 132-hydroxy-(132-S)-pheophytin a were significantly increased after stimulation, while IL-1α, IL-1β, IL-3, IL-12p70, and RANTES were unchanged.
Conclusions:
The investigation shows that components of L. leucocephala have actual cestocidal activity against H. nana.
... 9,10 A number of reports from plant extracts under in vitro and in vivo conditions have shown important anthelmintic effects on the GIN of livestock. 11,12 Lippia graveolens H.B.K is an aromatic plant in the Verbenaceae family, commonly named oregano in Mexico, and it is used as a condiment in Mexican food. This plant has been used as herbal remedies that are purported to be analgesic, antipyretic, antidiarrheal and antimalarial. ...
The objective of this study was to assay the ovicidal and larvicidal effects of two hydroalcoholic extracts (HA-E) and their aqueous fractions (Aq-F) and organic fractions (EtOAc-F) from Lippia graveolens (leaves) and Delonix regia (flowers) against the parasitic nematode Haemonchus contortus. The egg hatching inhibition test (% EHI) and mortality of infective larvae (% mortality) were used as biological models. The treatments of each plant (mg/mL, % EHI, and % mortality) were assigned as follows: HA-E (12.5–50, EHI; 25–200, mortality), Aq-F (6.25–25.00, EHI; 12.5–50, mortality), EtOAc-F (0.37–25, EHI; 3.12–50, mortality), negative controls (distilled water and methanol at 2% and 4%), and positive control (ivermectin at 5 mg/mL). Data were analyzed using a completely randomized design through an analysis of variance, considering the EHI and mortality percentages as the response variables. To identify the secondary metabolites, both plants were subjected to qualitative chemical analysis and HPLC. The results showed that both plants exhibited highly ovicidal and larvicidal effects. The bipartition in each HA extract allowed the increase of the anthelmintic effect in EtOAc-F, achieving a total ovicidal activity at 1.56 mg/mL and a larvicidal effect higher than 90% at 25 mg/mL. The qualitative chemical profile of both vegetal species identified alkaloids, flavonoids, condensed tannins, terpenes, and saponins. These results suggest that either L. graveolens leaves or D. regia flowers could be potential candidates for the control of parasitic gastrointestinal nematodes in small ruminants. However, in vivo studies with experimental animals to evaluate their possible toxicity and anthelmintic effect are necessary.
... On the other hand, recent studies have shown that extracts from the foliage of G. floribundum have anthelmintic (AH) activity in vitro against eggs and larvae of H. contortus (11,12) , and polyphenols have been shown to be involved in such activity (12) . In vitro AH activity was recently confirmed in in vivo studies using G. floribundum foliage in the diet of lambs infected with H. contortus (13) . ...
... On the other hand, recent studies have shown that extracts from the foliage of G. floribundum have anthelmintic (AH) activity in vitro against eggs and larvae of H. contortus (11,12) , and polyphenols have been shown to be involved in such activity (12) . In vitro AH activity was recently confirmed in in vivo studies using G. floribundum foliage in the diet of lambs infected with H. contortus (13) . ...
... The eggs were washed three times with purified water to remove the remaining sugar and were resuspended in 15 ml tubes containing 10 ml of phosphate saline solution (PBS 0.01 M: NaCl 0.138 M, KCl 0.0027 M, KH2PO4 0.001M, Na2HPO4 0.0081M; pH 7.4; Sigma® USA). Egg concentration was determined and the suspension was diluted to 150 eggs/ml of PBS for use in the egg hatch (EH) test (12) . ...
Se determinó el efecto del mes de cosecha y edad de las hojas de Gymnopodium floribundum sobre el contenido de compuestos polifenólicos (fenoles totales (FT), taninos totales (TT) y taninos condensados (TC)) de extractos metanol:agua. Además, se determinó la actividad biológica de los polifenoles medida como la capacidad de precipitar proteína (PP), inhibir la eclosión de huevos (EH), y el desenvaine larval (IDL) de Haemonchus contortus. Se cosecharon hojas de G. floribundum en cuatro meses del año: diciembre, marzo, junio y septiembre. Se obtuvieron 24 extractos metanol:agua (70:30), 12 producidos de hojas de edad variada (EV) y 12 de hojas de 90 días (E90). Todos los extractos ocasionaron similar PP independientemente de edad y mes de cosecha. La inhibición de la EH solo fue significativa para el extracto EV de diciembre (CE50 = 374.4 μg/ml; P<0.05). Los extractos de hojas E90 mostraron una CE50 > 1500 μg/ml en diciembre, junio y septiembre. Aunque todos los extractos inhibieron el desenvaine larval (IDL), la menor CE50 fue la del extracto de hojas EV de junio (CE50 = 80.4 μg/ml; P<0.05). La incubación de extractos con polivinilpolipirrolidona (PVPP) limitó la IDL (P<0.05), pero los polifenoles solo explicaron parte de esa actividad. En conclusión, el contenido de TC de los extractos de hojas de G. floribundum depende de su edad y mes de cosecha. Los polifenoles mostraron actividad de PP y se asociaron parcialmente con la IDL. Sin embargo, los polifenoles no explican la actividad contra huevos de H. contortus.
... Finally, H. albicans, G. floribundum, and S. gaumeri showed EC 50 values of 63.5, 66.9, and 184.7 µg/mL, respectively in the LEIT. Castañeda-Ramírez et al. [55] provided the first in vitro proposal to evaluate the nutraceutical value of the ten feed resources used in the previous experiment. In this protocol, the authors evaluated the macronutrient content, the in vitro dry matter, and the organic matter digestibility. ...
Nutraceuticals are defined as livestock feeds that combine their nutritional value with their beneficial effects on animal health. We analyzed the outcomes from nearly 20 years of research assessing the nutraceutical properties of plants consumed by sheep and goats in low deciduous forests. A systematic review of different databases suggested 31 peer-reviewed manuscripts according to pre-established criteria. Amongst these, 16 manuscripts described in vitro evaluations investigating the bioactivity of plant secondary compounds in the extracts of 12 plant species. Most of these studies used the abomasal nematode Haemonchus contortus as the parasite model. Meanwhile, 11 manuscripts reported in vivo trials under controlled pen conditions, evaluating the relationships between the intake of leaves from different plant species and their secondary compounds and animal nutrition, performance, and gastrointestinal nematode infections. Additionally, four manuscripts described studies under natural feeding conditions. Altogether, the studies showed the inherent complexity of the relationship between small ruminants, plants, nutrients, secondary compounds, and gastrointestinal nematodes in natural feeding systems. Several plant species can be considered good candidates for nutraceutical use. Our findings warrant future work to understand the relationship between plants, ruminants, and their parasites, with the aim to improve the sustainability of production systems based on the native vegetation of tropical forests.
... The anthelmintic (AH) activity of the plant secondary metabolites (PSM) contained in nutraceutical feedstuffs may help to reduce the dependence on commercial AH drugs at farm level [3]. Most nutraceutical feedstuffs evaluated for their AH activity against GIN under in vitro and in vivo conditions, either from temperate [4] or tropical regions [5,6], are normally consumed by small ruminants. The use of tanninrich nutraceutical legumes, such as Onobrychis viciifolia or Lespedeza cuneata fodders, as an aid for the management of GIN, are already used for small ruminants under different forms (i.e. ...
... Acetone was removed using a rotavapor (< 50 °C; Buchi model B-480) followed by two consecutive liquid-liquid extractions of the water fraction using dichloromethane (1:1) to remove pigments and lipids. This was roto-evaporated again to eliminate solvent residues [5]. Finally, the extract was lyophilized (Labconco UL Standard 61010 a ) and stored at 4 °C in sealed glass until use. ...
... Hence, if the agroindustrial by-product possess a good nutritional value and an AH activity against parasitic GIN, they may be considered as nutraceuticals [7]. The present study used the in vitro approach described by Castañeda-Ramírez et al. [5] to screen the nutraceutical potential of T. cacao husks and leaves of three varieties. ...
PurposeSome agroindustrial by-products could be used as nutraceutical materials for small ruminants helping with their nutrition while controlling their gastrointestinal nematodes. This study evaluated the potential in vitro nutraceutical value of pod husks and leaves of three varieties of Theobroma cacao using two Haemonchus contortus isolates with different polyphenol susceptibility.Methods
Leaves and husks from three T. cacao varieties (AZT, CAL and CEY) were evaluated for their bromatological composition, in vitro dry matter digestibility and polyphenol content. Acetone:water extracts (70:30) of each plant variety were evaluated using the egg hatch and larval exsheathment inhibition tests, using two isolates (FESC and PARAISO) of H. contortus. Effective concentrations 50% (EC50) were determined for both tests. The role of polyphenols was confirmed using polyvinylpolypyrrolidone. L3 exposed to CAL leaf extract were submitted to transmission electron microscopy.ResultsBoth plant materials showed a good nutritional value to complement protein-rich diets for small ruminants. Extracts inhibited exsheathment of H. contortus L3 more effectively than the egg hatching, and the leaf extracts were more active than husk extracts in the L3 exsheathment inhibition. The FESC isolate was more sensitive to extracts. Polyphenols blocked exsheathment inhibition of leaf extracts. Structural damage was observed in the sheath and muscles of L3 exposed to CAL leaf extracts.Conclusion
The two T. cacao materials tested showed their potential to be used as ruminant feeds. Extracts affected H. contortus by blocking L3 exsheathment, particularly with the leaf extracts. The in vivo nutraceutical value should be confirmed in small ruminants.
... When comparing the IC 50 values of L. leucocephala for LEI [50][51][52], found lower values than those observed in this study. In addition, the IC 50 of G. sepium for EHI in gastrointestinal sheep parasites [53] was lower than that observed here, and lower than that found [42], while a lower IC 50 was observed [48] when using extracts and fractions of cotyledon and L. leucocephala seeds. ...
In the present work, the anthelmintic activity (AA) of ethanolic extracts obtained from Gliricidia sepium, Leucaena leucocephala, and Pithecellobium dulce was evaluated using the third-stage-larval (L3) exsheathment inhibition test (LEIT) and egg hatch test (EHT) on Haemonchus contortus. Extracts were tested at concentrations of 0.3, 0.6, 1.2, 2.5, 5.0, 10, 20, and 40 mg/mL. The larval exsheathment inhibition (LEI) results showed that G. sepium achieved the highest average inhibition of 91.2%, compared with 44.6% for P. dulce and 41.0% for L. leucocephala at a concentration of 40 mg/mL; the corresponding IC50 values were 22.4, 41.7, and 43.3 mg/mL, respectively. The rates of egg hatching inhibition (EHI) at a concentration of 5 mg/mL were 99.5% for G. sepium, 64.2% for P. dulce, and 54% for L. leucocephala; the corresponding IC50 values were 1.9 mg/mL for G. sepium, 3.9 mg/mL for P. dulce, and 4.3 mg/mL for L. leucocephala. The species extracts studied here were also analyzed by ultra-high performance liquid chromatography and Orbitrap high resolution mass spectrometry (UHPLC-Q/Orbitrap/MS/MS), resulting in the compounds’ identification associated with AA. Glycosylated flavonoids and methoxyphenols were observed in all three species: fatty acids in G. sepium and P. dulce; phenylpropanoids, anthraquinone glycosides, amino acids and glycosylated phenolic acids in G. sepium; and flavonoids in L. leucocephala. Comparatively, G. sepium presented a greater diversity of compounds potentially active against the control of gastrointestinal nematodes, which was associated with the results obtained in the applied tests.
