Gastrointestinal Nematodes Of Sheep Research Articles

Infection with gastrointestinal nematodes in lambs in different integrated crop-livestock systems (ICL)

Lamb performance was evaluated in four integrated crop-livestock (ICL) systems. The ICL areas were previously planted with maize and marandu palisade grass in December 2013. ICL systems 1 and 2 were also seeded with pigeon pea. After harvesting in April 2014, black oats were sown in rows (ICL 1 and 3) or by broadcast seeding (ICL 2 and 4). Each ICL area was divided into 12 paddocks of 225 m2 to be grazed by young sheep from July 23 to September 30, 2014. To determine if the pasture remained contaminated by free-living stages of sheep gastrointestinal nematodes (GIN) after approximately ten months without animals in the area, 12 worm-free “tracer” lambs (n = 3/ ICL system) grazed each ICL pasture for 14 consecutive days in July 2015 and were later housed in pens, where they remained for another 14 days. The tracer lambs acquired no worm infections, demonstrating that the area was cleared after 300 days without animals. To evaluate GIN infection and uncastrated male lamb performance, we used 60 (n = 15/ system) Poll Dorset x Corriedale (crossbred) sheep with mean body weights of 24.4 ± 3.4 kg from July to September 2015 and 48 (n = 12/ system) Texel x Corriedale sheep with body weights of 26.4 ± 3.5 kg from June to September 2016. Lambs were allocated to the following groups: Group 1 rotated on the 12 ICL1 paddocks; Group 2 rotated on the 12 ICL2 paddocks; Group 3 rotated on the 12 ICL3 paddocks; and Group 4 rotated on the 12 ICL4 paddocks. Each paddock was grazed by sheep twice for three days, with a 33-day interval between grazing. At the end of the day the lambs were supplemented with concentrate plus silage. Groups 1 and 2 received mixed silage made of maize, marandu palisade grass and pigeon pea and groups 3 and 4 received mixed silage made of maize and marandu palisade grass. Faecal and blood samples were taken from all the animals every two weeks, and body weight was recorded on the same occasion. Nematode faecal egg counts (FEC), packed cell volume and total plasma protein means did not differ (P > 0.05) between the four lamb groups. In 2015, throughout the experimental period, the FEC decreased in all animals, with 6733 and 1407 eggs per gram (EPG) on average at the beginning and end of the trial, respectively. At the beginning of the trial in 2016, the animals had mild GIN infections (1077 EPG on average), and the individual faecal egg counts did not exceed 10,000 EPG during the trial. In decreasing order, Haemonchus spp., Trichostrongylus spp. and Cooperia spp. third stage larvae were found in the faecal cultures in both years. The performance of lambs were similar in the four ICL systems (P > 0.05), in the first and second years, the daily weight gains were 0.192 ± 0.05 kg and 0.221 ± 0.06 kg, respectively. In conclusion, a period of withdrawal of contaminated sheep results in plots free of infective larvae. The use of clean pastures during the dry season, in the different ICL systems, associated with a good nutrition plan, resulted in progressively declining degrees of GIN infections and satisfactory daily weight gain of young sheep.

Droplet digital polymerase chain reaction (ddPCR) as a novel method for absolute quantification of major gastrointestinal nematodes in sheep

In this paper, we present for the first time a new tool, based on Droplet Digital™ Polymerase Chain Reaction (ddPCR), for absolute quantification of key genera of gastrointestinal (GI) nematode parasites of grazing livestock. Four combinations of primers/probe sets targeting the internal transcribed spacer region 2 (ITS2) of the ribosomal RNA gene array were designed using the Primer3 software, following in silico analysis of nucleotide sequences from nematodes of interest downloaded from common databases. The amplified regions include both a universal region for detection of any strongylid gastrointestinal parasite and three different genus specific regions, making it possible to differentiate between the most important GI nematodes of sheep in Sweden: Haemonchus, Teladorsagia and Trichostrongylus. Analysis of samples containing serial dilutions and different mixtures of genomic DNA extracted from different species of adult worms proved useful in assessment of different threshold settings with the QuantaSoft software. Analysis of template DNA from these worms indicated that ddPCR is a viable choice for detection and absolute quantification of the different genera and also in samples with multiple species. Interpretation of the ddPCR results was straightforward and choice of analytical approach had little influence on the final results. Thus, the results obtained in the different analytical approaches seemed to be robust and the concentrations determined were uniform. Furthermore, the linear range of the Haemonchus ddPCR assay was similar to that of real-time PCR (qPCR). Taken together, our data confirm the suitability of ddPCR for detection and absolute quantification of three major sheep pathogens when tested on larval cultures from pooled ovine faeces. The results also indicate that ddPCR can be a useful complement to applications based on conventional egg counting methods such as the faecal egg reduction test (FECRT).

Efficacy of essential oils from plants cultivated in the Amazonian Biome against gastrointestinal nematodes in sheep.

The excessive use of anthelmintics to control nematodes has resulted in anthelminthic resistance. Essential oils (EOs) are a rich source of bioactive molecules that can be assessed for their ability to control resistant parasite populations. The aims of this study were to screen EOs from 10 plant species in vitro for anthelmintic activity against Haemonchus contortus, evaluate the cytotoxicity of those EOs in a human immortalized keratinocyte cell line (HaCaT), and test the most promising EO candidate in vivo in Santa Inês sheep. The efficacy was investigated in vitro using an egg hatch test (EHT) and a larval development test (LDT). EO cytotoxicity was evaluated with the sulforhodamine-B assay. In the in vivo experiment, 28 Santa Inês sheep naturally infected were distributed into groups: G1-Mentha arvensis (EO5), 200mgkg-1; G2-menthol, 160mgkg-1; G3-negative control; and G4-positive control (monepantel). EO5, from M. arvensis (86.7% menthol), had the lowest LC50 and LC90 values in the EHT (0.10, 0.27mgmL-1, respectively), good performance in the LDT (0.015, 0.072mgmL-1, respectively), and the lowest cytotoxicity (190.9µgmL-1) in HaCaT cells. In the in vivo test, a single dose of the EO5 (200mgkg-1 BW) had an efficacy of approximately 50% on days 1, 14, and 21; however, values were not significantly from day 0. Conversely, pure menthol at a dose of 160mgkg-1 BW showed no in vivo efficacy. This can be attributed to key factors related to bioavailability and pharmacology of terpenes in the host organism, as well as to the fact that menthol is mainly excreted as glucuronides in urine. Thus, further studies should be conducted with formulation systems that deliver bioactives directly to the abomasum, focusing on terpenes, whose excretion route is mainly via faeces.

Gastrointestinal nematodes in sheep of cutting under grazing regime

Sheep breeding is practiced in practically all the continents, the wide diffusion of the species is due mainly to its power of adaptation. Due to geoclimatic factors, tropical and subtropical countries stand out and settle in the agricultural scenario, and Brazil does not become an exception in this market. Brazil, because it has a considerable part of the herd produced in pasture, which is one of the economic and practical ways of producing and offering bulky ruminants, is one of the main meat producers at one of the lowest production costs. Tropical and subtropical climates, in addition to favoring optimal conditions for the production and development of agricultural and livestock sectors, such as pasture and ruminant farming, also provide favorable conditions for the development and survival of gastrointestinal nematodes. Negative performance of the animals, especially the sheep. Gastrointestinal nematodes, when uncontrolled, can cause 30 to 40% losses in animal performance, especially in the development of young animals and reproductive matrices that are the most sensitive categories, and which are closely related to the growth of the stock. The main nematodes that affect the small ruminants and of greater economic importance for the exploitation of sheep are: Haemonchus contortus and Trichostrongylus axei, both located in the abomasum; Trichostrongylus colubriformis, Strongyloides papillousus, Cooperia sp. and Bunostomum trigonocephalum, in the small intestine and Oesophagostomum columbianum, Trichuris ovis, T. globulosa and Skrjabinema sp., in the large intestine. Morphophysiological characteristics of forage plants such as foliar structure, habit of growth and green mass production, associated to climatic factors, may be factors that influence the permanence of the different stages of free life of the gastrointestinal nematodes in the environment, pastures that do not provide lightning in the lower stratum of the clumps and soils, provide shading and humidity and may potentiate the reinfection of the animals.

Effectiveness of Cratylia argentea as an animal feed supplement in the control of gastrointestinal nematodes in sheep

Gastrointestinal nematodes are considered one of the most important sanitary problems affecting ruminants globally; whether parasitizing alone or acting with other pathogens, they are responsible for severe economic losses related to animal husbandry. Endoparasites in small ruminants are usually controlled with anti-helminthic drugs. However, certain strains of endoparasites are resistant to such drugs. Supplementation of the diet with protein-rich foods is one strategy for reducing the negative impact of gastrointestinal parasites. Cratylia argentea has anthelmintic properties and other characteristics that favor its use as an animal feed supplement. In the present study, we subdivided crossbred lambs into homogeneous groups and fed them C. argentea (G1), C. argentea plus Brachiaria spp. (G2), or Brachiaria spp. (G3) only. The animals in group G1 had a lower average number of eggs per gram of feces (EPG) at Day 42 (p < 0.05), and the average count was ten times lower than that in the control at the last evaluated time-point (Day 56). The animals exhibited linear weight gain, and those in group G1 gained the most weight. The combined use of drugs and plants containing bioactive compounds can minimize the evolution of multidrug resistance, prolonging the efficacy of the drugs. The FAMACHA® score showed little variation during the experiment (p > 0.05), indicating the absence of anemia and normal globular volume. This reinforces the importance of the measurement of EPG; this is a quantitative method and does not depend on the subjective interpretation of the results.

Correction: The bacterial community associated with the sheep gastrointestinal nematode parasite Haemonchus contortus.

[This corrects the article DOI: 10.1371/journal.pone.0192164.].

Control of sheep gastrointestinal nematodes using the combination of Duddingtonia flagrans and Levamisole Hydrochloride 5.

The objective was to evaluate the action of D. flagrans pellets in association with Levamisole Hydrochloride 5% for controlling sheep gastrointestinal nematodes in the northeastern Brazil. Three groups of six sheep each were formed: group 1 received 3 g of the pellets (0.6 g of D. flagrans mycelium) for each 10 kg b.w., twice a week for six months, and deworming with Levamisole Hydrochloride 5% when EPG ≥ 1500; group 2 received a dosage of Levamisole Hydrochloride 5% when EPG ≥ 1500; and group 3 received 3 g of pellets without fungi for each 10 kg b.w., twice a week for six months. EPG counts, larval cultures, packed cell volume (PCV) and weighing were performed every 15 days; monthly, samples of grass from each paddock were collected. The mean EPG of the groups began to statistically differ from day 30 (p < 0.05). Group 1 required less deworming with Levamisole Hydrochloride 5% and showed superiority of PCV values ​​throughout the experiment (p < 0.05). There was a significant reduction (p < 0.05) in L3 recovery in the group 1 paddock from day 30 onwards. The use of D. flagrans pellets in association with Levamisole Hydrochloride 5% was effective for controlling gastrointestinal nematodes.

The bacterial community associated with the sheep gastrointestinal nematode parasite Haemonchus contortus.

Culture-independent methods were used to study the microbiota of adult worms, third-stage larvae and eggs, both in faeces and laid in vitro, of Haemonchus contortus, a nematode parasite of the abomasa of ruminants which is a major cause of production losses and ill-health. Bacteria were identified in eggs, the female reproductive tract and the gut of adult and third-stage larvae (L3). PCR amplification of 16S rRNA sequences, denaturing gradient gel electrophoresis (DGGE) and clone libraries were used to compare the composition of the microbial communities of the different life-cycle stages of the parasites, as well as parasites and their natural environments. The microbiomes of adult worms and L3 were different from those in the abomasum or faeces respectively. The H. contortus microbiota was mainly comprised of members of the phyla Proteobacteria, Firmicutes and Bacteroidetes. Bacteria were localised in the gut, inside eggs and within the uterus of adult female worms using the universal FISH Eub338 probe, which targets most bacteria, and were also seen in these tissues by light and transmission electron microscopy. Streptococcus/Lactococcus sp. were identified within the distal uterus with the probe Strc493. Sequences from the genera Weissella and Leuconostoc were found in all life-cycle stages, except eggs collected from faeces, in which most sequences belonged to Clostridium sp. Bacteria affiliated with Weissella/Leuconostoc were identified in both PCR-DGGE short sequences and clone libraries of nearly full length 16S rRNA bacterial sequences in all life-cycle stages and subsequently visualised in eggs by fluorescent in situ hybridisation (FISH) with group-specific probes. This strongly suggests they are vertically transmitted endosymbionts. As this study was carried out on a parasite strain which has been maintained in the laboratory, other field isolates will need to be examined to establish whether these bacteria are more widely dispersed and have potential as targets to control H. contortus infections.

Anthelminthic resistance of gastrointestinal nematodes in sheep to monepantel treatment in central region of Rio Grande do Sul, Brazil

ABSTRACT: Given the numerous reports of anthelminthic resistance of sheep nematodes to different anthelmintic compounds, this study aimed to evaluate the resistance status of gastrointestinal nematodes from naturally infected sheep to monepantel in the state of Rio Grande do Sul. Four farms that present extensive raising system and absence of anthelmintic treatment for 60 days were selected for the study. Lambs that present counts of eggs per gram of feces (EPG) ≥200 (sensitivity of 50 EPG) one day (D-1) before the treatment were select for the study and randomly separated into two groups, a control group and an experimental group treated with monepantel. Feces were collected 9 days after the treatment (D+9) for EPG counts and fecal culture. The monepantel was 100% effective only on 2. The efficacy found on farm 1, 3, and 4 were 2.82%, 25.8%, and 78.4%, respectably. There were no viable larvae post-treatment at farm 2, but the genera Haemonchus, Trichostrongylus, Cooperia, and Strongyloides were resistant to it at the other farms. This study shows the presence of parasites resistant to the treatment with monepantel, pointing to the importance of monitoring its efficacy in sheep flocks of Rio Grande do Sul, Brazil.

Prevalence of Gastrointestinal Nematodes in Sheep and Goats Reared by Nomads of Jammu Region of Jammu and Kashmir

The study was carried out to determine the prevalence of gastrointestinal nematodes in sheep and goats domesticated by nomads of Jammu region of J&K. A total of 283 faecal samples were collected and processed by centrifugal fecal flotation technique using 33% ZnSO4 flotation solution to ensure that the flotation is effective. 194 samples were found positive for different gastrointestinal nematodes. The study showed the prevalence rate of gastrointestinal nematodes in sheep and goats to be 65.04 % and 70.55% respectively. Haemonchus was found to be predominant in faecal samples of both sheep and goats. The young ones and females of both species showed the higher parasitic burden compared to adults and males respectively. Based on the findings of present study it is clear that sheep and goats of Jammu region reared by nomads are heavely infected with a variety of nematode parasites.

A Cross-sectional Study on the Prevalence and Associated Risk Factors of Gastrointestinal Nematodes of Sheep in and around Hirna, South East Ethiopia

A Cross-sectional Study on the Prevalence and Associated Risk Factors of Gastrointestinal Nematodes of Sheep in and around Hirna, South East Ethiopia

Prevalence and Associated Risk Factors of Gastrointestinal Nematodes of Sheep and Goats in ZiwayDugda District, Eastern Arsi Zone of Oromia Regional State, Ethiopia

Prevalence and Associated Risk Factors of Gastrointestinal Nematodes of Sheep and Goats in ZiwayDugda District, Eastern Arsi Zone of Oromia Regional State, Ethiopia

In vitro Anthelmintic Activity of Two Aloe-derived Active Principles against Sheep Gastrointestinal Nematodes

The in vitro anthelmintic activity on sheep gastrointestinal strongyle (GIS) eggs and larvae of 0.5% aloin and 0.1% aloe-emodin was investigated. From fresh faecal samples collected by ewes naturally infected by Haemonchus, Trichostrongylus and Teladorsagia nematodes, GIS eggs were isolated and cultivated in Petri dishes (100 eggs/dish). For the in vitro evaluation of the anthelmintic activity of tested compounds, the Egg hatch test (EHT), the Larval development test (LDT) and the Larval mortality/paralysis test (LMT) were used. In each assay, the activity of tested compounds was compared to untreated and treated (0.1% thiabendazole, TBZ) controls. Six repetitions were made through the experiment. Obtained data were statistically elaborated using the X2 test. In EHT, 0.5% aloin gave highly significantly different (P&lt;0.01) results from the untreated controls. In LDT, both 0.1% aloe-emodin and 0.5% aloin almost completely prevented the larval development from L1 to L3, showing no significant differences (P&lt;0.01) when compared to TBZ. In LMT, larval mortality observed in 0.5% aloin treated plates was significantly higher (P&lt;0.01) than that observed in TBZ treated controls. These results show the in vitro anthelmintic properties on sheep GIS of the examined plant secondary metabolites. In LDT and/or LMT, the activity of 0.5% aloin and 0.1% aloe-emodin was comparable to or higher than that of the reference drug.

Resistência anti-helmíntica em nematoides gastrintestinais de ovinos

The objective was to verify the existence of antihelminthic resistance in gastrointestinal nematodes of sheep in properties of the municipalities of Janaúba and Nova Porteirinha. The experiment was conducted at 4 properties, and was done as coproparasitological analyzes in fecal samples of 88 animals. In the farms (A and B) there was a division of the animals into two groups: control and immersion and in farms (C and D) in three groups: control, ivermectin and albendazole. Each group has 6 animals. The reduction of fecal grain contamination (RCOF) was determined based on the specific formula, indicating a percentage greater than or equal to 95%. There was resistance of the gastrointestinal nematodes to albendazole in the two properties in which it was tested and ivermectin in three of the four properties. In farm C, ivermectin showed efficacy of 97%. The most prevalent nematode was Haemonchus contortus.

Purification of native M. vogae and H. contortus tubulin by TOG affinity chromatography

Microtubules are non-covalent cylindrical polymers formed by alpha- and beta-tubulin heterodimer units, crucial for cell division, intracellular transport, motility and differentiation. This makes them very attractive pharmacological targets exploited to develop different drugs such as anthelmintics, antifungals, and antineoplastics. In this work, in order to establish an in vitro target-based screen to integrate to the search for new anthelmintics, we explored the extraction of native assembly-competent tubulin from two helminth parasites: Mesocestoides vogae tetrathyridia (syn. corti, Cestoda: Cyclophyllidea), a useful cestode biological model, and Haemonchus contortus, a sheep gastrointestinal nematode of interest in livestock production. For this purpose, a novel tubulin affinity chromatography procedure was employed, based on the binding capacity of TOG (Tumor Overexpressed Gene) domain from MAPs (microtubule-associated proteins). The TOG domain of the protein Stu2 from Saccharomyces cerevisiae fused to GST (glutathione S- transferase) were produced in E. coli, and the immobilized recombinant proteins allowed for native tubulin extraction from parasites. The binding capacity of TOG1 affinity column (3.6%) was estimated using commercial porcine brain tubulin. A total amount of up to 126 μg of M. vogae tubulin was purified, whereas H. contortus tubulin co-eluted with glutamate dehydrogenase enzyme. The identity of tubulins was confirmed by western blotting and mass spectrometry. The abundance of tubulin estimated in M. vogae was 10% soluble extract, which probably could explain differences observed between tubulin purification results of both helminth parasites.

Aspects of epidemiology and control of gastrointestinal nematodes in sheep and cattle – Approaches for its sustainability

Gastrointestinal helminthosis in ruminants cause considerable economic losses, with marked impairment of productivity of these animals, especially in poor management systems and poor parasite control programs. Infections caused by these helminths represent an obstacle to the expansion of the sheep industry. Furthermore, high parasite burdens were associated with increased mortality in calves. The recommended methods for the control of gastrointestinal helminths not always have practical applicability. We discuss important epidemiological and control aspects concerning gastrointestinal nematodes in sheep and cattle. Thus, sustainable worm control practices are recommended. Each farm has a specific situation and a management system directed to a specific parasitic infection must be adopted.

Anthelmintic effect of thymol and thymol acetate on sheep gastrointestinal nematodes and their toxicity in mice.

Thymol is a monoterpene and acetylation form of this compound can reduce the toxicity and enhance its biological effects. The objective of this study was to evaluate the effect of thymol and thymol acetate (TA) on egg, larva and adult Haemonchus contortus and the cuticular changes, acute toxicity in mice and the efficacy on sheep gastrointestinal nematodes. In vitro tests results were analyzed by analysis of variance (ANOVA) and followed by comparison with Tukey test or Bonferroni. The efficacy of in vivo test was calculated by the BootStreet program. In the egg hatch test (EHT), thymol (0.5 mg/mL) and TA (4 mg/mL) inhibited larval hatching by 98% and 67.1%, respectively. Thymol and TA (8 mg/mL) inhibited 100% of larval development. Thymol and TA (800 µg/mL) reduced the motility of adult worms, by 100% and 83.4%, respectively. Thymol caused cuticular changes in adult worm teguments. In the acute toxicity test, the LD50 of thymol and TA were 1,350.9 mg/kg and 4,144.4 mg/kg, respectively. Thymol and TA reduced sheep egg count per gram of faeces (epg) by 59.8% and 76.2%, respectively. In in vitro tests thymol presented better anthelmintic activity than TA. However TA was less toxic and in in vivo test efficacy was similar.

Nematophagous fungi combinations reduce free-living stages of sheep gastrointestinal nematodes in the field

Gastrointestinal nematodes (GIN) can reduce or limit sheep production. Currently there is a clear deficiency in the action of drugs for the control of these parasites. Nematophagous fungi are natural enemies of GIN. Fungal combinations have potential for reducing GIN populations. The aim of this study was to evaluate the efficiency combinations of nematophagous fungi in sodium alginate matrix pellets for the biological control agents of gastrointestinal sheep nematode parasites in the field. The nematophagous fungi (0.2mg of fungus per kg of body weight), Arthrobotrys conoides, A. robusta, Duddingtonia flagrans, and Monacrosporium thaumasium were used. The treated groups were administered mycelium combinations in the following combinations: group 1 (D. flagrans+A. robusta); group 2 (M. thaumasium+A. conoides). The control group did not receive any fungal pellets. We used three groups with eight Santa Inês sheep each. Each animal was treated with approximately 1g of pellet per 10kg of live weight. During the experimental period, we evaluated: number of eggs per gram of feces (EPG), infective larvae (L3) per kg of dry matter, larvae recovered from coprocultures, packed cell volume, total plasma protein concentration of sheep, and environmental conditions. Group 2 EPG (M. thaumasium+A. conoides) differed from the control group in September and October. The number of L3/kg of dry matter recovered from animals of groups 1 and 2 at distances of 0-20 and 20-40cm from the fecal pats was lower than the control group. The packed cell volume and total plasma proteins of treated animals were similar to those of the control group. The combination of treatment groups (D. flagrans+A. robusta and M. thaumasium+A. conoides) reduced the number of L3/kg of pasture. Therefore, treatment of nematophagous fungal combinations have the potential to manage free-living stages of GIN in sheep.

Evaluation of anthelmintic antimicrobial and antioxidant activity of Chenopodium album.

The objective of this study was to evaluate the anthelmintic, antimicrobial and antioxidant activities of Chenopodium album against gastrointestinal nematodes of sheep and some pathogenic microbes. A worm motility inhibition assay was used for in vitro study, and a faecal egg count reduction assay was used for an in vivo study. Various concentrations ranging from 100 to 500μg/ml of the extract were subjected to antimicrobial screening by disc diffusion method against four selected bacterial (Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas multocida and Escherichia coli) and two fungal (Aspergillus flavus and Candida albicans) strains in order to estimate the medicinal potential of the herb. DPPH (1,1-diphenyl-2-picrylhydrazyl), riboflavin photo-oxidation, deoxyribose, lipid peroxidation assays were used for antioxidant activity. The extracts exhibited dose- and time-dependent anthelmintic effects on the Haemonchus contortus as compared to levamisole. The extract showed maximum inhibitory effect against S. aureus (28±0.14mm), while as mild inhibitory effect was observed against E. coli among the selected microbial strains. The effect produced by the different extract concentrations was comparable with the standard antibacterial agent streptomycin sulphate and antifungal agent nystatin, which were used as effective positive control in the study. The antioxidant activity showed that the extracts exhibited scavenging effect in concentration-dependent manner on superoxide anion radicals and hydroxyl radicals leading to the conclusion that the plant has broad spectrum anthelmintic, antimicrobial and antioxidant activities and could be a potential alternative for treating various diseases.

Synergistic interaction of ten essential oils against Haemonchus contortus in vitro

Anthelmintic resistance in sheep gastrointestinal nematodes is a worldwide problem. Multi-drug resistant haemonchosis is the most serious impediment for small ruminant systems, and there are no new drug candidates currently under development. Molecules from natural sources have demonstrated anthelmintic activity against parasites. In this work, the monoterpenoids carvacrol, carvone, cineole, linalool, limonene, and thymol and the phenylpropanoids cinnamaldehyde, anethole, vanillin, and eugenol were assessed individually or in mixtures of ten binary, three ternary, and three quaternary combinations using the in vitro egg hatch assay with eggs of a multi-drug resistant strain of Haemonchus contortus. The main objective of this study was to identify the most effective interaction among essential oils with the greatest individual anthelmintic efficacy and to determine the most powerful combinations. The essential oils were ranked by their 50% lethal concentration (LC50) as follows (mg/mL): cinamaldehyde (0.018), anethole (0.070), carvone (0.085), carvacrol (0.11), thymol (0.13), linalool (0.29), vanillin (0.57), eugenol (0.57), cineole (4.74), and limonene (207.5). Quantification of synergism, additive effect, and antagonism were calculated for binary, ternary, and quaternary combinations. The best anthelmintic effect resulting from synergistic activity among 16 different combinations was for cinnamaldehyde:carvacrol (CL50 0.012mg/mL) and anethole:carvone (CL50 0.013mg/mL). These results indicate that these binary combinations would be promising to be tested in sheep infected with H. contortus.