Egg Hatching Inhibition Research Articles

Nematicidal Efficacy of Milbemectin against Root-Knot Nematodes.

The nematicidal efficacy of milbemectin and its commercial formulate Milbeknock® on (i) egg hatching, (ii) juvenile motility and (iii) infective capacity of root-knot nematodes was evaluated in vitro and in planta assays. Serial dilutions of pure milbemectin were tested against nematode eggs and juveniles and lethal concentrations LC50 and LC90 calculated. Exposure of egg masses to milbemectin at a concentration of 30 μg/mL for 72 h reduced egg hatching by 52%. The increase in exposure time to 240 h did not increase the egg hatching inhibition at the highest concentration 30 μg/mL (53%) but reduced egg hatching at 15 and 7 μg/mL by 35 and 24%, respectively, when compared to untreated controls. The inhibitory effect of milbemectin on juvenile motility ranged from 41 to 87% depending on its concentration, and this effect was persistent after rinsing the juveniles in water. The probabilistic dose–response model indicated that lethal concentrations of milbemectin for juvenile motility were LC50: 7.4 μg/mL and LC90: 29.9 μg/mL. The pre-plant application of Milbeknock® to soils infested with the nematode reduced its infective capacity by 98–99% compared to untreated soils in pot experiments. Milbeknock® reduced nematode soil population densities by 50–60% in natural infestations under field conditions. Milbemectin shows a high level of efficacy against root-knot nematodes as it reduces egg hatching, persistently immobilizes nematode juveniles, and reduces tomato root infection.

In-vitro efficacy of Arachis hypogaea (Peanut) peels extract against Haemonchus contortus

Synthetic anthelmintics are becoming ineffective due to development of parasitic resistance. For this reason, traditional medicinal plants are being explored for their anthelmintic potential. The aim of this study was to evaluate the in-vitro anthelmintic activity of Arachis hypogaea L. (peanut) peels against Haemonchus contortus. To assess the anthelmintic effect of Arachis hypogaea L. on 3 life cycles of Haemonchus contortus, egg hatch assay (EHA), larval developmental assay (LDA) and adult motility assay (AMA) were conducted. In these tests, methanol and n-hexane extracts of the plant were used in three concentrations of 10, 15 and 20 mg/ml. Levamisole and PBS were used as positive and negative control groups respectively. Results of these tests showed that methanol extract of Arachis hypogaea L. had higher anthelmintic effect than that of n-hexane extract. Overall both extracts exhibited a significant (p<0.05) dose and a time dependent anthelmintic effect. At 20 mg/ml, methanol extract and n-hexane extract showed 87% and 80% egg hatching inhibition respectively. Methanol and n-hexane extracts at 20 mg/ml showed 83.3% and 76.6% larval mortality respectively. Adult motility test with both extracts showed maximum immobilization of worms after 6 hours of treatment at 20 mg/ml concentration. It is concluded that peels of Arachis hypogaea possess significant anthelmintic potential against nematodes. It may be suggested that the plant can be used further to investigate the in-vivo activity.

In Vitro Anthelmintic Evaluation of Gliricidia sepium, Leucaena leucocephala, and Pithecellobium dulce: Fingerprint Analysis of Extracts by UHPLC-Orbitrap Mass Spectrometry.

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.

In Vitro Screening of a Culturable Soybean Cyst Nematode Cyst Mycobiome for Potential Biological Control Agents and Biopesticides.

Fungal biological control of soybean cyst nematodes (SCN) is an important component of integrated pest management for soybean. However, very few fungal biological control agents are available in the market. In this study, we have screened fungi previously isolated from SCN cysts over 3 years from a long-term crop rotation field experiment for their ability to antagonize SCN using (i) parasitism, (ii) egg hatch inhibition, and (iii) J2 mortality. We evaluated egg parasitism using an in-vitro egg parasitism bioassays and scored parasitism using the egg parasitic index (EPI) and fluorescent microscopy. The ability of these fungi to produce metabolites causing egg hatch inhibition and J2 mortality was assessed in bioassays using filter-sterilized culture filtrates. We identified 10 high-performing isolates each for egg parasitism and toxicity toward SCN eggs and J2s and repeated the tests after storage for 1 year of cryopreservation at -80°C to validate the durability of biocontrol potential of the chosen 20 isolates. Although the parasitic ability changed slightly for the majority of strains after cryopreservation, they still scored 5/10 on EPI scales. There were no differences in the ability of fungi to produce antinemic metabolites after cryopreservation.[Formula: see text] Copyright © 2020 The Author(s). This is an open access article distributed under the CC BY 4.0 International license.

Screening of anti-nematode potential through inhibition of egg hatching in plant-parasitic nematode Meloidogyne javanica

Plant-parasitic nematodes have emerged as nature’s most successful among all parasites known till today. These animals have been reported from all terrains of all ecosystems. Their capability to survive on a wide diversity of the host plants, circumvent host plant defence is a few of several of their secrets making them most successful of all known parasites. Among various groups of plant-parasitic nematodes, endo-parasitic nematodes are the most damaging one and also difficult to control. Meloidogyne sps. are commonly known as root-knot nematodes. Our inability to control them is primarily due to our poor understanding of the biology of these plant parasites. Due to the availability of the complete genome sequence of few Meloidogyne species, biotechnological interventions are used to unravel the secrets of their success. Chemical controls of these nematodes are extensively reported in the literature. Due to the environmental toxicity associated with these chemicals, and restrictions on the use of chemicals against nematodes led to screening and development of eco-friendly management strategies. The present study was conducted to screen nematotoxic properties of Neem (Azadirachta indica), Jatropha (Jatropha curcas), Kachnar (Bauhinia variegate), Bel (Aegle marmelos) and Eucalyptus (Eucalyptus globules) leaf extracts against root-knot nematode Meloidogyne javanica in vitro. The aqueous extracts were used against the hatching of the nematode eggs, movement of second stage juveniles (J2) and the viability of the J2 in increasing concentration of the bioactive compound. The eggs were treated with various concentrations of the selected extracts for different time periods ranging from 24h to 6 days. A significant inhibition of egg hatching and increase in the mortality of the nematode juvenile in few of the aqueous extracts were recorded. Reduced egg hatching and increased mortality of the nematode juveniles could be maybe the indicators of the presence of anti-nematode potential in the selected plant leaves. The results from the study can pave the way for the development of eco-friendly management strategies for plant-parasitic nematodes.

Artemisia cina 30 CH homeopathic treatment against Haemonchus contortus

The anthelmintic resistance problem is widely recognized in sheep production. Therefore, new methods of control against gastrointestinal nematodes (GIN) need to be integrated. The aim of this work was to assess the toxicity of A. cina 30 CH as a homeopathic product against Haemonchus contortus in in vitro and in vivo assays. A. cina 30 CH was obtained from a commercial laboratory, and confirmation of artemisinin as a key ingredient was performed with mass spectrophotometry. The A. cina 30 CH and the artemisinin pure reagent were used for the inhibition of egg hatching (IEH) and for the inhibition of larval migration of H. contortus L3 (ILM). In addition, three groups of 10 naturally infected lambs with GIN were treated with A. cina 30 CH and albendazole, and 10 were used as control. The parasitic infection was monitored at 0, 7, 14 and 28 d postreatment (PT) to determine the number of eggs per gram (epg) and FAMACHA index. The in vitro data showed 100 % IEH and 64.7 % ILM by A. cina 30 CH, and nonlethal activity was observed with the artemisinin pure reagent. The toxicity of A. cina 30 CH against H. contortus in infected lambs was observed after 7 d of infection. Administration of the A. cina 30 CH yielded a 69 % reduction in the epg at 28 d PT, similar to the albendazole (P<0.05). In conclusion, A. cina 30 CH had the ability to IEH and ILM of H. contortus in in vitro assays and reduced the number of eggs of H. contortus, which is the primary parasitic nematode in grazing lambs, thereby reducing infection.

Nematicidal Effect of Methyl Palmitate and Methyl Stearate against Meloidogyne incognita in Bananas.

Banana plants (Musa spp.) are susceptible to infection by many plant-parasitic nematodes, including Meloidogyne incognita. In this study, a mixed fermentation broth of chicken manure (CM) and cassava ethanol wastewater (CEW) was used to inhibit M. incognita by reducing egg hatching and by having a lethal effect on second-stage juvenile nematodes (J2s). It also alleviated nematode damage and promoted banana plant growth. Using gas chromatography-mass spectrometry (GC-MS), we identified methyl palmitate and methyl stearate as bioactive compounds. These bioactive compounds repelled J2s and inhibited egg hatching; reduced root galls, egg masses, and nematodes in soil; and downregulated the essential parasitic nematode genes Mi-flp-18 and 16D10. A Caenorhabditis elegans offspring assay showed that low concentrations of the fermentation broth, methyl palmitate, and methyl stearate were safe for its life cycle. This study explored the effective and environmentally safe strategies for controlling root-knot nematodes.

In vitro evaluation of Penicillium chrysogenum Snef1216 against Meloidogyne incognita (root-knot nematode)

Root-knot nematode (Meloidogyne incognita) is chief plant parasitic nematode of various crops globally. Meanwhile, the negative side effects on human health and environmental concerns associated with haphazard uses of chemical nematicides. Hence, the search for a safe and effective approach is more relevant. The present study was aimed to evaluate the nematicidal potential of Snef1216 (Penicillium chrysogenum) against M. incognita at different concentrations (5%, 10%, 25%, 50%, 75% and 100%) and with the nutritious medium. The egg hatching inhibition and mortality of second stage juveniles of M. incognita were assessed after 6, 12, 24, 48 and 72 h exposure. Results revealed that egg hatching inhibition and percent mortality of M. incognita increased with increasing concentration and exposure time. The highest mortality of juveniles was recorded at 100% conc. i.e., 24.20%, 36%, 66%, 78% and 97.8% at 6, 12, 24, 48 and 72 h, respectively. The highest ovicidal activity was recorded at 100% concentration with 5.20% of eggs hatching. The outcome suggested that Snef1216 (P. chrysogenum) resulted in the lowest LC50 value was recorded as 3718.496 at 6 h exposure period followed by 10479.87, 11186.682, 14838.58 and 24001.430 at 72, 12, 48 and 24 h respectively via ovicidal assay. Whereas, in the larvicidal assay, the lowest LC50 value demonstrated at 72 h being 17.628% exposure period followed by 28.345, 50.490, 215.710 and 482.595% at 48, 24, 12 and 6 h respectively. It is concluded that Snef1216 has potential being used as a biocontrol agent against M. incognita and can serve as a source of a novel nematicidal agent of fungal origin.

Activity of four nematicides against Meloidogyne incognita race 2 on tomato plants

AbstractMeloidogyne incognita is one of the most important causes of disease in protected vegetable cultivation in North China Plain, but chemical control options for it are currently limited. In the present study, we measured activity of four nematicides with M. incognita race 2 in the laboratory and greenhouse pots. Fluensulfone and fluopyram had a greater negative effect on the motility of M. incognita second‐stage juveniles (J2) than avermectin B1a (AV‐B1a) and fosthiazate had, especially at low concentration, with respective EC50 values of 0.29, 0.13, 0.68 and 2.48 mg/L. AV‐B1a significantly and uniquely inhibited egg hatching, which indicated that it was the only nematicide that could readily be transported across the eggshell. In greenhouse pots, fluensulfone (10 mg−1) and fluopyram (1 and 10 mg−1) caused the greatest inhibition of formation of galls in tomato roots, with decreases of 98.6%, 96.2% and 99.2%, respectively. This indicated that some M. incognita J2 lost their root penetration ability without losing their motility.

In vitro Anthelmintic Activity of Aqueous and Ethanolic Extract of Senna italica (Caesalpiniaceae) on Three-stages of Haemonchus contortus

A phytochemical screening and in vitro anthelminthic activity of aqueous and ethanolic extract of Senna italica on Haemonchus contortus were conducted. Polyphenol, tannin and flavonoid contents were determined by using gallic acid and rutin. egg hatching inhibition test was carried out on fresh eggs; larval mortality test was conducted on infective larvae (L3) and adult worm mortality test was conducted. Eggs, larvae and adults worms were incubated in aqueous and ethanolic extract of S. italica at different concentrations (0.1; 0.3; 0.5; 0.7 and 1 mg/mL). All extracts of plants showed an effect on all stages of H. contortus with high efficiency variations depending on the dose used. The inhibition of eggs hatching rate increased from 8.67±1.53% to 65.67±1.15% and from 24.67±1.53% to 80±1.73% respectively for aqueous and ethanolic extract of S. italic. The larval mortality rate increased from 12.22±2.34% to 56.67±4.9% after 24 h and from 45.28±4.11% to 91.25±3.73% after 48 h for aqueous extract and from 30.07±2.84% to 70.83±6.31% after 24 h and from 48.79±3.73% to 96.25±4.79% after 48 h for ethanolic extract. After 24 h of exposure to aqueous and ethanolic extract of S. italica, the adult mortality rate varies from 55.56±9.62% to 83.33±9.62% for aqueous extract and from 61.11±9.62% to 88.89±9.62%.These in vitro results confirm the use of S. italica in traditional medicine.

Identification and Characterization of Nematicidal Volatile Organic Compounds from Deep-Sea Virgibacillus dokdonensis MCCC 1A00493.

Root-knot nematode diseases cause severe yield and economic losses each year in global agricultural production. Virgibacillus dokdonensis MCCC 1A00493, a deep-sea bacterium, shows a significant nematicidal activity against Meloidogyne incognita in vitro. However, information about the active substances of V. dokdonensis MCCC 1A00493 is limited. In this study, volatile organic compounds (VOCs) from V. dokdonensis MCCC 1A00493 were isolated and analyzed through solid-phase microextraction and gas chromatography–mass spectrometry. Four VOCs, namely, acetaldehyde, dimethyl disulfide, ethylbenzene, and 2-butanone, were identified, and their nematicidal activities were evaluated. The four VOCs had a variety of active modes on M. incognita juveniles. Acetaldehyde had direct contact killing, fumigation, and attraction activities; dimethyl disulfide had direct contact killing and attraction activities; ethylbenzene had an attraction activity; and 2-butanone had a repellent activity. Only acetaldehyde had a fumigant activity to inhibit egg hatching. Combining this fumigant activity against eggs and juveniles could be an effective strategy to control the different developmental stages of M. incognita. The combination of direct contact and attraction activities could also establish trapping and killing strategies against root-knot nematodes. Considering all nematicidal modes or strategies, we could use V. dokdonensis MCCC 1A00493 to set up an integrated strategy to control root-knot nematodes.

In Vitro Anthelmintic Activity of Crude Extracts of Artemisia herba-alba and Punica granatum against Haemonchus contortus.

Gastrointestinal nematodes (GINs) are the major limiting factor for the successfulness of livestock production throughout the world. Emergence of resistance strains as well as scarcity and high cost of the currently available drugs has led to the evaluation of other alternative helminth control options, mainly from plants. The current study is aimed at investigating the in vitro anthelmintic efficacy of crude methanolic extracts of two traditionally important medicinal plants, Artemisia herba-alba and Punica granatum, against Haemonchus contortus using adult motility assay (AMA) and egg hatch inhibition assay (EHIA). Four graded concentrations of the extracts were tested for both the AMA (10, 5, 2.5, and 1.25 mg/mg) and EHIA (0.1, 0.25, 0.5, and 1 mg/mL) in replicates. Albendazole and phosphate-buffered saline (AMA) or distilled water (EHIA) were used as the positive and negative controls, respectively. The crude extracts of A. herba-alba and P. granatum exhibited a potential anthelmintic activity at all dose levels in a concentration- and time-dependent fashion. The highest concentration (10 mg/mL) of all the extracts caused a significantly (p < 0.05) superior nematocidal activity compared to the negative control. Moreover, significant and concentration-dependent egg hatching inhibition effect was observed from both plant extracts. Maximal (98.67%) egg hatching inhibition effect was exhibited by the flower extract of A. herba-alba at 1 mg/mL concentration. The relative egg hatch inhibition efficacy indicated that both plants caused a significantly (p < 0.05) greater egg hatch inhibition within 48 hr of exposure. The current study validated the traditional use of both plants as a natural anthelmintic against H. contortus justifying a need to undertake detail pharmacological and toxicological investigation on both plants.

Synthesis and Evaluation of Pyrazole Derivatives as Potent Antinemic Agents

Pyrazole derivatives were synthesized by bromination of pyrazole, followed by N-alkylation of 4-bromopyrazole. The synthesized derivatives were characterized by microanalytical data and IR and 1H and 13C NMR spectra and were evaluated for their nematicidal activity against the root knot nematode Meloidogyne incognita. The compounds were screened for their egg hatch inhibition and mortality potential, and they showed significant nematicidal activity as compared to the control. 1H-Pyrazol-5(4H)-one was found to be most effective in egg hatch inhibition, and 4-bromopyrazole was found to be most effective in juvenile mortality.

Biopesticidal value of Senecio glaucus subsp. coronopifolius essential oil against pathogenic fungi, nematodes, and mites

Essential oils are widely investigated as an alternative or complementary approach to broad-spectrum synthetic pesticides. In the present study, essential oil of Senecio glaucus subsp. coronopifolius was screened for its antifungal, nematicidal, acaricidal and repellent activities against the phytopathogens Botrytis cinerea, Meloidogyne javanica and Tetranychus urticae Koch under laboratory conditions. The volatile phase method (VF) and poisoned food method (PF) were adopted to test the antifungal activity of the essential oil. The oil gave 83% inhibition at 16 μL/ml using the PF method. VF method showed 86% inhibition of mycelial growth of B. cinerea at 0.8 μL/ml air. To check the nematicidal activity of the essential oil, two bioassays targeted the mortality of second stage juvenile (J2) and hatch inhibition of M. javanica eggs. 95% immobility of J2 and 92% inhibition of egg hatch were recorded at 16,000 ppm. The effect exerted by the oil was nematostatic. In the case of mites, a leaf dip bioassay revealed 100% mortality of T. urticae adults, and repellency of 24% after exposure to the oil at 80% concentration. Chemical profile of S. glaucus essential oil was obtained using GC–MS. Terpenic compounds, such as indicated α-pinene (26.2%), myrcene (11.4%), p-cymene (9.9%), β-pinene (7.7%), γ-muurolene (4%), deoxynivalenol (3.1%) and α-phellandrene (2.7%) were identified as the major components. The findings indicated that S. glaucus essential oil has the potential for management of these pathogens.

Nematicidal potential of nematophagous fungi on Meloidogyne incognita

The efficacy of indigenous fungi isolated from mite,grub,teabark and soil sample were tested against Meloidogyne incognita for their efficacy in vitro through egg hatch inhibition, J2 mortality and egg parasitism. Mortality of M. incognita larvae was found to be directly proportional to concentration of culture filtrate of the isolates and duration of exposure in vitro. Among the isolate listed, Isolate 8 showed highest egg hatch inhibition, juvenile mortality and egg parasitism of M. incognita. followed by Isolate 10, Isolate 1 and Isolate 2.

Festulolium and fungal endophyte associations: host status for Meloidogyne incognita and nematotoxic plant extracts.

Festulolium hybrids are forage grasses used worldwide in temperate climates. They are associated with the fungal endophyte Epichloë uncinata, which aids in nutrient uptake, drought tolerance, and production of metabolites that protect against parasites and herbivores. Epichloë uncinata produces loline alkaloids, which can deter insect pests. Festulolium has not been widely studied for susceptibility to plant-parasitic nematodes, so Festulolium lines, with and without fungal endophytes, were tested in the greenhouse for host status to the root-knot nematode Meloidogyne incognita. All were poor hosts, regardless of line or endophyte status. Pepper seedlings planted into soil following removal of the Festulolium plants were infected by nematodes, likely because of surviving nematodes from the original inoculation combined with some reproduction on Festulolium. Lolines were found in shoots and roots of all endophyte-associated lines, and some types of lolines in roots increased after nematode infection. Methanolic extracts from roots and shoots of a tested Festulolium line did not inhibit egg hatch, but killed nearly a third of second-stage juveniles whether an endophyte was present or not. Further studies would indicate whether these Festulolium lines aid in suppressing field populations of M. incognita.

Egg hatch inhibition of root-knot nematodes by Tagetes patula extracts

Egg hatch inhibition of root-knot nematodes by <i>Tagetes patula</i> extracts

Cell-wall degrading enzymes for the control of Meloidogyne javanica

The potential of cell-wall degrading enzymes (chitinase, β-1,3 glucanase, endo-cellulase and exocellulase) in the control of M. javanica in vitro and in vivo was investigated in this study. The enzymes were produced by solid-state fermentation using Beauveria bassiana, Trichoderma harzianum and Metarhizium anisopliae, microorganisms that are used in the biological control of agricultural pests. The bioproducts were efficient in the treatments in vitro, presenting high indexes of egg hatching inhibition (66.7-87.0%) and mortality of juveniles (40.0-90.0%). The bioproduct containing enzymes from B. bassiana presented the most promising results. In vivo treatments with all bioproducts on lettuce and tomato cultures during initial stages of plant development demonstrated excellent nematicidal effects in relation to the water-treated control. On lettuce, final egg counts and juveniles in the control treatment were 141 and 452% higher than in treatments containing bioproducts, respectively. The index of penetration of M. javanica on tomato roots was 348% higher in the control treatment than in the treatments using bioproducts. The bioproducts evaluated in this study were efficient in the control of nematodes both in treatments in vitro and in vivo. The results obtained open a new perspective for the use of bioproducts containing cell-wall degrading enzymes for the control of nematodes in agriculture.

In vitro anthelmintic activity of Lippia gracilis Schauer essential oil against egg-hatching of goat gastrointestinal nematodes

ABSTRACT Since drug-resistant nematodes became a common problem in sheep and goat industries, alternative methods using natural products have emerged as a viable and sustainable anthelmintic treatment option. Here, the in vitro effect of essential oil extracted from Lippia gracilis Schauer was assessed on the hatching process of nematodes recovered from naturally infected goats. Essential oil at concentrations of 0.08% (0.008 μL/mL), 0.12% (0.012 μL/mL), and 0.16% (0.016 μL/mL) was able to induce an average inhibition of 74.7, 84 and 93%, respectively. The effective concentration required to inhibit egg hatching in 50% of eggs (EC50) was 0.03452%. Therefore, essential oil of L. gracilis showed promisor in vitro anthelmintic results against egg-hatching of goat gastrointestinal nematodes.

In vitro nematocidal activity of commercial fatty acids and β-sitosterol against Haemonchus contortus.

Haemonchus contortus is a haematophagous gastrointestinal nematode (GIN) that causes severe anaemia and even death in small ruminants, negatively impacting the economic viability of farms. Traditionally, this parasite has been controlled with chemical compounds; however, inadequate use of these types of products has favoured the emergence of anthelmintic resistance. Therefore, it is necessary to search for alternatives for GIN control. Previous studies have reported the anthelmintic activity of edible mushroom extracts against H. contortus. A recent study reported that a fraction constituted of different fatty acids and β-sitosterol isolated from the basidiomata of the edible mushroom Pleurotus djamor ECS-123 has ovicidal and larvicidal activity against H. contortus. Thus, this study aimed to assess the anthelmintic activity of the pure molecules: pentadecanoic acid, palmitic acid, β-sitosterol, stearic acid and linoleic acid. For this purpose, an egg-hatching inhibition test was carried out in which the compounds were evaluated individually and in combination at a final concentration of 20 mg mL-1. Furthermore, larval mortality was assessed using a combination of the five commercial compounds previously mentioned at different concentrations (1.25-20 mg mL-1). Palmitic acid and stearic acid, in some combinations, inhibited H. contortus egg hatching by 100%. On the other hand, in the larval mortality test, the combination of the five compounds showed dose-dependent behaviour, and 100% mortality was obtained 24 h post-incubation. Pure molecules and their combinations have anthelmintic-like activity against the eggs and larvae of H. contortus.