Control Of Parasitic Nematodes Research Articles

Chlamydospore dormancy and predatory activity of nematophagous fungus Duddingtonia flagrans.

The chlamydospores of Duddingtonia flagrans are an essential survival and reproductive structure and also an effective ingredient for the biocontrol of parasitic nematodes in livestock. In this study, entering and exiting dormancy conditions and predatory activity of the fungal chlamydospores were conducted. During this fungal growth process, the cultivation time is negatively correlated with spore germination rates. After the spores were processed by vacuum drying for 168 h, their germination rate dropped to 0.94%. In contrast, the percentage of living spores remained 54.82%, suggesting that the spores entered structural dormancy in the arid environment. Meanwhile, the efficacies of the spore against Haemonchus contortus larvae were 93.05% (0 h), 92.19% (16 h), 92.77% (96 h), and 86.45% (168 h), respectively. After dormant spores were stored at 4°C, -20°C, and 28°C (RH90 ~ 95%) for 7 days, their germination rate began to increase significantly (p < 0.05). For in vitro predation assay under the condition of 28°C (RH90 ~ 95%), the predation rate was significantly higher on the 7th day after incubation than that on the 3rd day (p < 0.05). During the period when spores were stored at room temperature for 8 months, their germination rate decreased in the first 5 months and then increased slowly to reach a peak in the 7th month. However, the reduction rate of H. contortus L3 in feces captured by spores remained above 71% for the first 7 months. These results will help us increase the end products yield and the quality of biological control of parasitic nematodes in livestock.

Selective control of parasitic nematodes using bioactivated nematicides.

Parasitic nematodes are a major threat to global food security, particularly as the world amasses 10 billion people amid limited arable land1-4. Most traditional nematicides have been banned owing to poor nematode selectivity, leaving farmers with inadequate means of pest control4-12. Here we use the model nematode Caenorhabditis elegans to identify a family of selective imidazothiazole nematicides, called selectivins, that undergo cytochrome-p450-mediated bioactivation in nematodes. At low parts-per-million concentrations, selectivins perform comparably well with commercial nematicides to control root infection by Meloidogyne incognita, a highly destructive plant-parasitic nematode. Tests against numerous phylogenetically diverse non-target systems demonstrate that selectivins are more nematode-selective than most marketed nematicides. Selectivins are first-in-class bioactivated nematode controls that provide efficacy and nematode selectivity.

The Brown Alga Bifurcaria bifurcata Presents an Anthelmintic Activity on All Developmental Stages of the Parasitic Nematode Heligmosomoides polygyrus bakeri.

The current control of gastrointestinal (GI) parasitic nematodes mainly relies on the widespread use of anthelmintics, which has inevitably led to resistance. Therefore, there is an urgent need to find new sources of antiparasitic compounds. Macroalgae represent a rich source of active molecules and are widely described as having medicinal properties. In the present study, we investigated the potential anthelmintic activity of aqueous extracts from three species of algae (Bifurcaria bifurcata, Grateloupia turuturu and Osmundea pinnatifida) on the murine parasite Heligmosomoides polygyrus bakeri. Using a set of complementary in vitro tests, including larval development assays, egg hatching tests and nematicidal activity assays on larvae and adults, we report the nematicidal activity of aqueous extracts of B. bifurcata. In addition, aqueous extract fractionation using liquid/liquid partitioning with a solvent of increasing polarity was performed in order to identify the groups of active molecules underlying the anthelmintic activity. Non-polar extracts (heptane, ethyl acetate) demonstrated high anthelmintic potential, highlighting the role of non-polar metabolites such as terpenes. Here, we highlight the strong anthelmintic potential of the brown alga B. bifurcata on a mouse model of GI parasites, thus confirming the strong interest in algae as natural alternatives for the control of parasitic nematodes.

Tuft Cells Increase Following Ovine Intestinal Parasite Infections and Define Evolutionarily Conserved and Divergent Responses.

Helminth parasite infections of humans and livestock are a global health and economic problem. Resistance of helminths to current drug treatment is an increasing problem and alternative control approaches, including vaccines, are needed. Effective vaccine design requires knowledge of host immune mechanisms and how these are stimulated. Mouse models of helminth infection indicate that tuft cells, an unusual type of epithelial cell, may ‘sense’ infection in the small intestine and trigger a type 2 immune response. Currently nothing is known of tuft cells in immunity in other host species and in other compartments of the gastrointestinal (GI) tract. Here we address this gap and use immunohistochemistry and single cell RNA-sequencing to detail the presence and gene expression profile of tuft cells in sheep following nematode infections. We identify and characterize tuft cells in the ovine abomasum (true stomach of ruminants) and show that they increase significantly in number following infection with the globally important nematodes Teladorsagia circumcincta and Haemonchus contortus. Ovine abomasal tuft cells show enriched expression of tuft cell markers POU2F3, GFI1B, TRPM5 and genes involved in signaling and inflammatory pathways. However succinate receptor SUCNR1 and free fatty acid receptor FFAR3, proposed as ‘sensing’ receptors in murine tuft cells, are not expressed, and instead ovine tuft cells are enriched for taste receptor TAS2R16 and mechanosensory receptor ADGRG6. We also identify tuft cell sub-clusters at potentially different stages of maturation, suggesting a dynamic process not apparent from mouse models of infection. Our findings reveal a tuft cell response to economically important parasite infections and show that while tuft cell effector functions have been retained during mammalian evolution, receptor specificity has diverged. Our data advance knowledge of host-parasite interactions in the GI mucosa and identify receptors that may potentiate type 2 immunity for optimized control of parasitic nematodes.

Phylum-Spanning Neuropeptide GPCR Identification and Prioritization: Shaping Drug Target Discovery Pipelines for Nematode Parasite Control.

Nematode parasites undermine human health and global food security. The frontline anthelmintic portfolio used to treat parasitic nematodes is threatened by the escalation of anthelmintic resistance, resulting in a demand for new drug targets for parasite control. Nematode neuropeptide signalling pathways represent an attractive source of novel drug targets which currently remain unexploited. The complexity of the nematode neuropeptidergic system challenges the discovery of new targets for parasite control, however recent advances in parasite ‘omics’ offers an opportunity for the in silico identification and prioritization of targets to seed anthelmintic discovery pipelines. In this study we employed Hidden Markov Model-based searches to identify ~1059 Caenorhabditis elegans neuropeptide G-protein coupled receptor (Ce-NP-GPCR) encoding gene homologs in the predicted protein datasets of 10 key parasitic nematodes that span several phylogenetic clades and lifestyles. We show that, whilst parasitic nematodes possess a reduced complement of Ce-NP-GPCRs, several receptors are broadly conserved across nematode species. To prioritize the most appealing parasitic nematode NP-GPCR anthelmintic targets, we developed a novel in silico nematode parasite drug target prioritization pipeline that incorporates pan-phylum NP-GPCR conservation, C. elegans-derived reverse genetics phenotype, and parasite life-stage specific expression datasets. Several NP-GPCRs emerge as the most attractive anthelmintic targets for broad spectrum nematode parasite control. Our analyses have also identified the most appropriate targets for species- and life stage- directed chemotherapies; in this context we have identified several NP-GPCRs with macrofilaricidal potential. These data focus functional validation efforts towards the most appealing NP-GPCR targets and, in addition, the prioritization strategy employed here provides a blueprint for parasitic nematode target selection beyond NP-GPCRs.

Evaluation of cross-grazing deer with sheep or cattle, as means to reduces anthelmintic usage to control gastrointestinal and pulmonary nematodes in farmed red deer (Cervus elaphus) in New Zealand

Recent reports indicate that gastrointestinal nematodes (GIN) are contributing to significant losses in deer productivity and that anthelmintic resistance has become an issue of concern for deer farmers in New Zealand. The aim of this study was to evaluate cross-grazing of deer with sheep or cattle as an aid for control of gastrointestinal and pulmonary nematode parasites of farmed red deer (Cervus elaphus) in New Zealand. This was a field study replicated over two years (2012 and 2013) for 16 weeks each year at two locations (Massey University, Palmerston North and Invermay AgResearch Centre, Mosgiel). Each farm replicate included four groups (19–20 deer) at each location: red deer cross-grazing with cattle (Deer/Cattle); red deer cross-grazing with sheep (Deer/Sheep); red deer grazing on their own (DeerOwn); and red deer grazing on their own and treated with anthelmintics every two weeks to suppress worm burdens, as a positive control (DeerSup). The key outcome was the number of anthelmintic treatments (AT) given to deer. The decision to treat individual resident deer in Deer/Cattle, Deer/Sheep and DeerOwn groups was based on “trigger” criteria including faecal egg count (FEC)≥250 eggs/g or Dictyocaulus faecal larval count (FLC)≥100 larvae/g or when growth rate was less than 80 % of the mean of the DeerSup group in the previous two weeks. In addition, to quantify the species of parasites cycling in each group, sets of three “tracer” deer were introduced to graze with each group at the mid-point and again at the end of each 16 week period in both years at both locations. Least squares means (LSM) of the number of AT given per animal for Deer/Sheep (3.4) and DeerOwn (3.3) groups were significantly higher than for the Deer/Cattle (2.7) group (p < 0.001). In tracer animals, the LSM of abomasal Trichostrongylus spp. were significantly fewer in the DeerOwn (17), Deer/Cattle (37) and DeerSup (54) groups than in the Deer/Sheep (952; p < 0.001) group. The LSM of the nematodes in the subfamily Ostertagiinae (=Ostertagia-type) were significantly more in the DeerOwn (1950) than in Deer/Sheep (370; p = 0.003) and DeerSup (238; p < 0.001) groups, but the number in the Deer/Cattle group (689) was not different to DeerOwn (p> 0.05). The LSM of lungworm were fewer in Deer/Sheep (3), Deer/Cattle (4) and DeerSup (3; p < 0.001) groups than in DeerOwn (40) group. The Deer/Cattle and DeerSup groups had significantly higher LSM of liveweight gain over the 16 weeks (p < 0.001) than the other two groups. This study demonstrated that cross-grazing with either sheep or cattle aided control of lungworm and gastrointestinal nematodes in young deer during autumn. However, the advantages varied between the use of sheep or cattle and in the ability to control different species of parasites.

Cytoplasmic Lipases-A Novel Class of Fungal Defense Proteins Against Nematodes.

Fungi are an attractive food source for predators such as fungivorous nematodes. Several fungal defense proteins and their protective mechanisms against nematodes have been described. Many of these proteins are lectins which are stored in the cytoplasm of the fungal cells and bind to specific glycan epitopes in the digestive tract of the nematode upon ingestion. Here, we studied two novel nematotoxic proteins with lipase domains from the model mushroom Coprinopsis cinerea. These cytoplasmically localized proteins were found to be induced in the vegetative mycelium of C. cinerea upon challenge with fungivorous nematode Aphelenchus avenae. The proteins showed nematotoxicity when heterologously expressed in E. coli and fed to several bacterivorous nematodes. Site-specific mutagenesis of predicted catalytic residues eliminated the in-vitro lipase activity of the proteins and significantly reduced their nematotoxicity, indicating the importance of the lipase activity for the nematotoxicity of these proteins. Our results suggest that cytoplasmic lipases constitute a novel class of fungal defense proteins against predatory nematodes. These findings improve our understanding of fungal defense mechanisms against predators and may find applications in the control of parasitic nematodes in agriculture and medicine.

Repeated, drug-truncated infections with Ostertagia ostertagi elicit strong humoral and cell-mediated immune responses and confer partial protection in cattle

Bovine ostertagiasis causes significant production losses to the cattle industry. Protective immunity induced by natural infection is slow to develop and anthelmintic resistance is rapidly developing. There is a need to advance alternatives for control of gastrointestinal nematode parasites. The present study investigated the effects of repeated, drug-truncated infections (rDTI) on development of protective immunity and attenuation of a challenge infection by O. ostertagi. Helminth-free calves were randomly assigned to either a rDTI or a control group (n = 5). The rDTI group received daily oral infections of 5000 Ostertagia L3 for 5 consecutive days, then were drug-treated on 14 and 15 days post infection (dpi), to attenuate O. ostertagi at the late fourth larval (L4) through young adult stages. DTI was repeated 3 weeks after the drug treatment. A total of 5 DTIs were administered to the DTI-treated animals. Non-DTI-treated, control animals received tap water as infection control. All animals were drug-treated at the same time. Animals were challenge-infected 4 weeks following the final round of rDTI. The results show that eggs per gram of feces (EPG) in the rDTI group were significantly reduced (P < 0.05) from 21 to 39 dpi, with an overall reduction in cumulative EPG. The control group exhibited reduced (P = 0.0564) average weight gains when compared to those of the rDTI group during weeks 4–5 post infection, a period coinciding with peak EPG output of control animals. Antigen-specific IgG, IgE and IgA responses were detected after the 2nd DTI, and stronger antibody recall responses were elicited by challenge infection. High levels of antigen-specific peripheral blood mononuclear cell (PBMC)/T cell proliferation to whole worm and excretory-secretory (ES) antigens were detected in rDTI-treated animals. These data indicate that partial protective immunity against ostertagiasis, involving cell-mediated and humoral responses, can be attained by rDTI which allowed for maximal antigen exposure from staggered parasitic developmental stages. The data suggest that rDTI can be used as a model to study host-parasite interactions and identify parasite antigens responsible for eliciting host protective immune responses.

The role of Cinnamomum zeylanicum essential oil, (E)‐cinnamaldehyde and (E)‐cinnamaldehyde oxime in the control of Meloidogyne incognita

AbstractThe search for natural nematicides that are biodegradable with little or no human toxicity has intensified in recent years. In this context, the use of essential oils has the potential to function as an alternative plant–parasitic nematode control strategy, and their characterization may identify new nematicidal molecules. In this study, the nematicidal activity of Cinnamomum zeylanicum essential oil, its most abundant biochemical component and its analogue were evaluated against Meloidogyne incognita. Mean LC50 and LC95 values for C. zeylanicum oil were 49 µg/ml and 131 µg/ml, respectively. When J2 placed in C. zeylanicum oil at its LC50 concentration were used to inoculate tomato plants, the reduction in numbers of galls and eggs versus samples inoculated using J2 and no C. zeylanicum oil was 98%. C. zeylanicum essential oil reduced levels of M. incognita J2 that hatched 38%–54%, while carbofuran (positive control) did not prevent hatching. C. zeylanicum oil applied at a concentration of 800 µg/ml to a potted substrate containing infested tomato plants significantly reduced numbers of M. incognita galls. The cinnamaldehyde molecule within C. zeylanicum oil had LC50 and LC95 values of 64 and 768 µg/ml, respectively, while LC50 and LC95 values for cinnamaldehyde oxime were 323 and 529 µg/ml, respectively. Both cinnamaldehyde and its oxime inhibited hatching in M. incognita J2. These findings indicate that C. zeylanicum essential oil, its major biochemical component, cinnamaldehyde and cinnamaldehyde oxime (a cinnamaldehyde analogue) can be used to reduce levels of M. incognita.

Chemogenomic approach to identifying nematode chemoreceptor drug targets in the entomopathogenic nematode Heterorhabditis bacteriophora

Parasitic nematodes constitute one of the major threats to human health, causing diseases of major socioeconomic importance worldwide. Recent estimates indicate that more than 1 billion people are infected with parasitic nematodes around the world. Current measures to combat parasitic nematode infections include anthelmintic drugs. However, heavy exposure to anthelmintics has selected populations of livestock parasitic nematodes that are no longer susceptible to the drugs, rendering several anthelmintics useless for parasitic nematode control in many areas of the world. The rapidity with which anthelmintic resistance developed in response to these drugs suggests that increasing the selective pressure on human parasitic nematodes will also rapidly generate resistant worm populations. Therefore, development of new anthelmintics is of major importance before resistance becomes widespread in human parasitic nematode populations. G-Protein Coupled Receptors (GPCRs) represent an important target for many pharmacological interventions due to their ubiquitous expression in various cell types. GPCRs contribute to numerous physiological processes, and their ligand binding sites located on cell surfaces make them accessible targets and attractive substrates in terms of druggability. In fact, ∼35 % of Food and Drug Administration (FDA) and European Medicines Agency (EMA) approved drugs target GPCRs and their associated proteins, with over 300 additional drugs targeting GPCRs at the clinical trial stage. Nematode Chemosensory GPCRs (NemChRs) are unique to nematodes, and therefore represent ideal substrates for target-based drug discovery. Here we set out to identify NemChRs that are transcriptionally active inside the host, and to use these NemChRs in a reverse pharmacological screen to impede parasitic development. Our data identified several NemChRs, and we focused on one that was expressed in neuronal cells and exhibited the highest fold change in transcription after host activation. Next, we performed homology modelling and molecular dynamics simulations of this NemChR in order to conduct a virtual screening campaign to identify candidate drug targets which were ranked and selected for experimental testing in bioassays. Taken together, our results identify and characterize a candidate NemChR drug target, and provide a chemogenomic pipeline for identifying nematicide substrates.

Identification of compounds responsible for the anthelmintic effects of chicory (Cichorium intybus) by molecular networking and bio-guided fractionation

Increasing resistance towards anthelmintic drugs has necessitated the search for alternative treatments for the control of gastrointestinal nematode parasites. Animals fed on chicory (Cichorium intybus L.), a temperate (pasture) crop, have reduced parasite burdens, hence making C. intybus a potentially useful source for novel anthelmintic compounds or a diet-based preventive/therapeutic option. Here, we utilized in vitro bioassays with the parasitic nematode Ascaris suum and molecular networking techniques with five chicory cultivars to identify putative active compounds. Network analysis predicted sesquiterpene lactones (SL) as the most likely group of anthelmintic compounds. Further bioassay-guided fractionation supported these predictions, and isolation of pure compounds demonstrated that the SL 8-deoxylactucin (8-DOL) is the compound most strongly associated with anti-parasitic activity. Furthermore, we showed that 8-DOL acts in a synergistic combination with other SL to exert the anti-parasitic effects. Finally, we established that chicory-derived extracts also showed activity against two ruminant nematodes (Teladorsagia circumcincta and Cooperia oncophora) in in vitro assays. Collectively, our results confirm the anti-parasitic activity of chicory against a range of nematodes, and pave the way for targeted extraction of active compounds or selective breeding of specific cultivars to optimize its future use in human and veterinary medicine.

The narrow-spectrum anthelmintic oxantel is a potent agonist of a novel acetylcholine receptor subtype in whipworms.

In the absence of efficient alternative strategies, the control of parasitic nematodes, impacting human and animal health, mainly relies on the use of broad-spectrum anthelmintic compounds. Unfortunately, most of these drugs have a limited single-dose efficacy against infections caused by the whipworm, Trichuris. These infections are of both human and veterinary importance. However, in contrast to a wide range of parasitic nematode species, the narrow-spectrum anthelmintic oxantel has a high efficacy on Trichuris spp. Despite this knowledge, the molecular target(s) of oxantel within Trichuris is still unknown. In the distantly related pig roundworm, Ascaris suum, oxantel has a small, but significant effect on the recombinant homomeric Nicotine-sensitive ionotropic acetylcholine receptor (N-AChR) made up of five ACR-16 subunits. Therefore, we hypothesized that in whipworms, a putative homolog of an ACR-16 subunit, can form a functional oxantel-sensitive receptor. Using the pig whipworm T. suis as a model, we identified and cloned a novel ACR-16-like subunit and successfully expressed the corresponding homomeric channel in Xenopus laevis oocytes. Electrophysiological experiments revealed this receptor to have distinctive pharmacological properties with oxantel acting as a full agonist, hence we refer to the receptor as an O-AChR subtype. Pyrantel activated this novel O-AChR subtype moderately, whereas classic nicotinic agonists surprisingly resulted in only minor responses. We observed that the expression of the ACR-16-like subunit in the free-living nematode Caenorhabditis elegans conferred an increased sensitivity to oxantel of recombinant worms. We demonstrated that the novel Tsu-ACR-16-like receptor is indeed a target for oxantel, although other receptors may be involved. These finding brings new insight into the understanding of the high sensitivity of whipworms to oxantel, and highlights the importance of the discovery of additional distinct receptor subunit types within Trichuris that can be used as screening tools to evaluate the effect of new synthetic or natural anthelmintic compounds.

Exposure to the anthelmintic dinitroaniline oryzalin causes changes in meiotic prophase morphology and loss of synaptonemal complexes in the nematode Caenorhabditis elegans

Abstract The anthelmintic dinitroaniline oryzalin interferes with the formation of microtubules and inhibits meiosis and mitosis in nematodes. Exposure to oryzalin resulted in deterioration in morphology of the oocytes and loss of synaptonemal complexes at meiotic prophase I. The nuclear matrix and envelope were poorly formed, and the central rachis was diminished. These results provide the basis for the loss of fecundity after treatment with the oryzalin resulting in control of parasitic nematodes.

Preference in Dogs of Two Oral Endectoparasiticide Formulations: NexGard Spectra&amp;lt;sup&amp;gt;&amp;#174;&amp;lt;/sup&amp;gt; (Afoxolaner and Milbemycin Oxime) and Credelio&amp;lt;sup&amp;gt;&amp;#174;&amp;lt;/sup&amp;gt; Plus (Lotilaner and Milbemycin Oxime)

Fleas and ticks are major ectoparasites of dogs globally. Their control is based on regular treatments with ectoparasiticides, which represent the most important part of veterinary drugs worth around 3 billion Euros per year. In many parts of the world, dogs are also at risk of infection by endoparasites like heartworm, eyeworm, and lungworm. In these areas, endectoparasiticide formulations are used to prevent the risk of ecto- and endoparasite infections. Since 2014, oral formulations of insecticidal-acaricidal drugs have been launched, followed by endectoparasiticide formulations. These oral formulations facilitate the treatment by the owners and are now the market leaders. Intense work has been done during their development to enhance their palatability through their consistence (hard to soft) and their flavors. Palatable oral formulations facilitate the dog-owner relationship and help ensuring compliance. The most recent palatable formulations include isoxazoline as the ecto-parasiticide molecule. They also include anthelmintics (milbemycin oxime or moxidectin + pyrantel) to provide control of parasitic nematodes. Being very similar in terms of spectrum of activity, any differences in palatability may be a key differentiating factor for the owners. The present study was conducted to verify if dogs exhibited a preference between two endectoparasiticide oral formulations, NexGard Spectra&#174 (afoxolaner and milbemycin oxime) and Credelio&#174 Plus (lotilaner and milbemycin oxime). For four consecutive days, 100 dogs were offered the choice between both products and consumption was recorded. If one product was more consumed than the other, it was defined as the preferred product. A total of 375 chewable tablets were consumed over the four days, with a significantly higher consumption (p 0.0001) for NexGard Spectra&#174 (272 chews, 72.5%) compared to Credelio&#174 Plus (103 chews, 27.5%). Seventy-six dogs showed a preference for a product amongst whom 68 preferred NexGard Spectra&#174 (89.5%) and 8 preferred Credelio&#174 Plus (10.5%), resulting in a preference ratio of 8.5 to 1 for NexGard Spectra&#174 (p 0.0001).

Announcement of WormAtlas partnership with the Journal of Nematology.

A detailed understanding of nematode anatomy can be leveraged for the development of new parasitic nematode control strategies and for fundamental biological insights through nematode model organisms. The Center for C. elegans Anatomy, with its websites WormAtlas and WormImage, is the central anatomical resource for researchers studying the model organism Caenorhabditis elegans. Here, we announce our expansion of the WormAtlas and WormImage resources beyond C. elegans to include additional nematode species. Towards this goal, we will partner with the Journal of Nematology to write and solicit anatomically focused review chapters for publication in the Journal and corresponding inclusion on the WormAtlas website.

Contrasting patterns of isotype-1 β-tubulin allelic diversity in Haemonchus contortus and Haemonchus placei in the southern USA are consistent with a model of localised emergence of benzimidazole resistance

The benzimidazoles are one of the most important broad-spectrum anthelmintic drug classes for parasitic nematode control in domestic animals and humans. They have been widely used in livestock, particularly in small ruminants for over 40 years. This has resulted in widespread resistance in small ruminant gastrointestinal nematode parasite species, especially Haemonchus contortus. Benzimidazole resistance mutations have also been reported in Haemonchus placei, but only at low frequencies, suggesting resistance is at a much earlier stage of emergence than is the case for H. contortus. Here, we investigate the haplotype diversity of isotype-1 β-tubulin benzimidazole resistance mutations and the population genetic structure of H. contortus and H. placei populations from sheep and cattle from the southern USA. Microsatellite genotyping revealed a low level of genetic differentiation in six H.placei and seven H. contortus populations examined. This is consistent with several previous studies from other regions, mainly in H. contortus, supporting a model of high gene flow between parasite populations. There was a single F200Y(TAC) haplotype present in all six H. placei populations across Georgia, Florida and Arkansas. In contrast, there were at least two different F200Y(TAC) haplotypes (up to four) and two different F167Y(TAC) haplotypes across the seven H. contortus populations studied. These results provide further evidence to support a model for benzimidazole resistance in Haemonchus spp, in which resistance mutations arise from a single, or the small number of locations, in a region during the early phases of emergence, and subsequently spread due to animal movement.

The Possible Biotechnological Use of Edible Mushroom Bioproducts for Controlling Plant and Animal Parasitic Nematodes.

The present paper reviewed publications on the nematocidal activity of edible mushrooms (EM) and their potential use as sustainable tools for the control of parasitic nematodes affecting agriculture and livestock industry. Nematodes are organisms living in the soil and animals' guts where they may live as parasites severely affecting economically important crops and farm animals, thus causing economic losses to worldwide agriculture. Traditionally, parasitic nematodes have been controlled using commercial pesticides and anthelmintic (AH) drugs. Over the years, nematodes developed resistance to the AH drugs, reducing the usefulness of many commercial drugs. Also, the use of pesticides/anthelmintic drugs to control nematodes can have important negative impacts on the environment. Different EM have been not only used as food but also studied as alternative methods for controlling several diseases including parasitic nematodes. The present paper reviewed publications from the last decades about the nematocidal activity of EM and assessed their potential use as sustainable tools for the control of nematodes affecting agriculture and livestock industry. A reduced number of reports on the effect of EM against nematodes were found, and an even smaller number of reports regarding the potential AH activity of chemical compounds isolated from EM products were found. However, those studies have produced promising results that certainly deserve further investigation. It is concluded that EM, their fractions and extracts, and some compounds contained in them may have biotechnological application for the control of animal and plant parasitic nematodes.

In vitro biological control of bovine parasitic nematodes by Arthrobotrys cladodes, Duddingtonia flagrans and Pochonia chlamydosporia under different temperature conditions.

Variations in temperature can affect the development of nematophagous fungi, especially when they are used in the biological control of parasitic nematodes in the pastures where cattle are reared. The aim of this work was to evaluate the effects of temperature on the performance of nematophagous fungi in the biological control of bovine parasitic nematodes. The mycelial growth, chlamydospore production and nematicidal activity of Duddingtonia flagrans, Arthrobotrys cladodes and Pochonia chlamydosporia were evaluated at 15, 20, 25, 30 and 35°C. The fungal strains achieved mycelial growth, chlamydospore production and nematicidal activity on parasitic nematodes under all temperature conditions tested. The fungi showed higher growth at intermediate temperatures (20, 25 and 30°C) than at the extremes of 15 and 35°C. At 25 and 30°C, D. flagrans realized 96.8 and 94.5% nematicidal activity on bovine parasitic nematodes, respectively. Arthrobotrys cladodes effected nematicidal activity of 85.3 and 83.5%, at 20 and 25°C, respectively. At 20 and 30°C, P. chlamydosporia achieved nematicidal activity of 81.3 and 87.4%, respectively. The maximum chlamydospore production was reached at 20, 25 and 30°C for D. flagrans, at 20 and 25°C for A. cladodes and P. chlamydosporia. The results of this study demonstrated that the tested fungal strains of D. flagrans, A. cladodes and P. chlamydosporia, when used in the biological control of bovine parasitic nematodes, were not limited by in vitro temperature variations. Therefore, the use of these strains of fungi as biological control agents of parasitic nematodes is promising.

Biological Control of Gastrointestinal Helminthes of Livestock

Gastrointestinal helminths cause considerable production loss in ruminants globally and constraint to optimum livestock production. Extensively managed animals often host gastrointestinal parasites. Guidelines for sustainable control of parasites in sheep (SCOPS) have been produced in the UK which include recommendations for treatments to ewes. Traditionally control has been achieved through the application of broad spectrum anthelmintic; however, resistance is the major issue. In terms of parasitic nematodes in sheep, widespread resistance has been identified to the three available broad-spectrum anthelmintic classes with reports of increasing multi-class resistance was reported. Anthelmintic treatments can reach up to eight for lamb finishing. Resistance to anthelmintic can be reached to 88%. The trend towards nonchemical parasite control methods (ecological, organic, green) farming of livestock. Biological control of nematode parasites of livestock is the rising issue of animal science. In this study, biological control of nematode parasites of livestock has been reviewed.

The transcription factor NHR-8: A new target to increase ivermectin efficacy in nematodes.

Resistance to the anthelmintic macrocyclic lactone ivermectin (IVM) has a great impact on the control of parasitic nematodes. The mechanisms by which nematodes adapt to IVM remain to be deciphered. We have identified NHR-8, a nuclear hormone receptor involved in the xenobiotic response in Caenorhabditis elegans, as a new regulator of tolerance to IVM. Loss-of-function nhr-8(ok186) C. elegans mutants subjected to larval development assays and electropharyngeogram measurements, displayed hypersensitivity to IVM, and silencing of nhr-8 in IVM-resistant worms increased IVM efficacy. In addition, compared to wild-type worms, nhr-8 mutants under IVM selection pressure failed to acquire tolerance to the drug. In addition, IVM-hypersensitive nhr-8(ok186) worms displayed low transcript levels of several genes from the xenobiotic detoxification network and a concomitant low Pgp-mediated drug efflux activity. Interestingly, some pgp and cyp genes known to impact IVM tolerance in many nematode species, were down regulated in nhr-8 mutants and inversely upregulated in IVM-resistant worms. Moreover, pgp-6 overexpression in nhr-8(ok186) C. elegans increased tolerance to IVM. Importantly, NHR-8 function was rescued in nhr-8(ok186) C. elegans with the homolog of the parasitic nematode Haemonchus contortus, and silencing of Hco-nhr-8 by RNAi on L2 H. contortus larvae increased IVM susceptibility in both susceptible and resistant H. contortus isolates. Thus, our data show that NHR-8 controls the tolerance and development of resistance to IVM in C. elegans and the molecular basis for this relates to the NHR-8-mediated upregulation of IVM detoxification genes. Since our results show that Hco-nhr-8 functions similarly to Cel-nhr-8, this study helps to better understand mechanisms underlying failure in drug efficacy and open perspectives in finding new compounds with NHR-8 antagonist activity to potentiate IVM efficacy.