Control Of Parasitic Nematodes Research Articles

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.

POTENCY OF LOCAL ISOLATE PAECILOMYCES FROM WEST SUMATERA FOR CONTROL OF ROOTKNOT NEMATODES (MELOIDOGYNE SPP) ON VEGETABLES

ABSTRACT Paecilomyces is an antagonistic fungus that can be used as a base material for the manufacture of bionematicides for the control of root knot nematodes (Meloidogyne spp) because it has antagonistic activity as a nematode egg parasite. Utilization of Paecilomyces local isolates has good potentials for the control of parasitic nematodes, especially root knot nematodes. The environmental conditions of fungal isolates affect the ability of fungi in parasitizing nematode eggs. The objective of the study was to obtain local isolate of Paecilomyces fungi which have high potential to control root knot nematodes. Fungal samples were from soil rhizosphere of tomato plants infected by root knot nematodes (Meloidogyne spp) from several locations of vegetable production centers of West Sumatra, namely Alahan Panjang, Agam and Tanah Datar. As results of the research, 10 isolates of Paecilomyces fungi were obtained, 3 isolates from Alahan Panjang (PAP1, PAP2, and PAP3), 3 isolates from Agam (PA1, PA2, and PA3) and 4 isolates from Tanah Datar (PTD1, PTD2. PTD3 and PTD4). All Paecilomyces isolates had potency to control root knot nematode (Meloidogyne spp) with suppression percentage ranged from 55% - 70.9%. The PAP 2 (isolate from Alahan Panjang) had the best ability to control root knot nematodes with suppression percentage of 70.90%.

Nicotine-sensitive acetylcholine receptors are relevant pharmacological targets for the control of multidrug resistant parasitic nematodes

The control of parasitic nematodes impacting animal health relies on the use of broad spectrum anthelmintics. However, intensive use of these drugs has led to the selection of resistant parasites in livestock industry. In that respect, there is currently an urgent need for novel compounds able to control resistant parasites. Nicotine has also historically been used as a de-wormer but was removed from the market when modern anthelmintics became available. The pharmacological target of nicotine has been identified in nematodes as acetylcholine-gated ion channels. Nicotinic-sensitive acetylcholine receptors (N-AChRs) therefore represent validated pharmacological targets that remain largely under-exploited. In the present study, using an automated larval migration assay (ALMA), we report that nicotinic derivatives efficiently paralyzed a multiple (benzimidazoles/levamisole/pyrantel/ivermectin) resistant field isolate of H. contortus. Using C. elegans as a model we confirmed that N-AChRs are preferential targets for nornicotine and anabasine. Functional expression of the homomeric N-AChR from C. elegans and the distantly related horse parasite Parascaris equorum in Xenopus oocytes highlighted some striking differences in their respective pharmacological properties towards nicotine derivative sensitivity. This work validates the exploitation of the nicotine receptors of parasitic nematodes as targets for the development of resistance-breaking compounds.

Ovicidal and larvicidal activity of extracts from medicinal-plants against Haemonchus contortus

The use of medicinal plants (MP) containing bioactive compounds is an alternative strategy to control of parasitic nematode of small ruminants Haemonchus contortus at various stages of their life cycle. The aims of this study were to determine the in vitro anthelmintic activity of both aqueous and methanolic extracts from 13 medicinal plants typical for Central Europe, and to determine quantity of selected plant secondary metabolites (PSMs) in the methanolic extracts. In vitro egg hatch test and larval development tests were conducted to determine the possible anthelmintic effects of methanolic and aqueous extracts of the roots of Althaea officinalis L., Petasites hybridus L. and Inula helenium L.; flowers of Malva sylvestris L. and Chamomilla recutita L.; leaves of Plantago lanceolata L. and Rosmarinus officinalis L.; seeds of Foeniculum vulgare Mill. and stems of Solidago virgaurea L., Fumaria officinalis L., Hyssopus officinalis L., Melisa officinalis L. and Artemisia absinthium L. on eggs and larvae of H. contortus. Ultra-performance liquid chromatography and tandem mass spectroscopy was used for quantifying six PSMs: gallic acid (GA), rutin (RU), diosmin (DI), hesperidin (HE), quercetin (QU) and kaempferol (KA). RU content of the most effective methanolic extracts was in the order: M. sylvestris (9.33 mg/g DM) > A. absinthium (6.10 mg/g DM) > C. recutita (0.42 mg/g DM). The highest concentration of QU (44.8 mg/g DM) and KA (6.59 mg/g DM) were detected in stems of F. officinalis comparing to the other evaluated plants. The most significant (p < 0.05) anthelmintic effects exhibited methanolic extracts of A. absinthium in both in vitro tests (i.e., egg hatch test and larval development test). Additionally, only two methanolic extracts of C. recutita and M. sylvestris were comparable to activity of A. absinthium using the larval development test. Wider spectrum of aqueous extracts exhibited stronger ovicidal activity in comparison to methanolic extracts. The similar trend was observed in evaluating of larvicidal activity of aqueous and methanolic plant extracts.

Biocontrol of Meloidogyne incognita by Metarhizium guizhouense and its protease

Crude extracts of the fungus Metarhizium guizhouense PSUM02 grown on protein- enriched medium showed high proteolytic activity and produced one main protein indicated as fungal protease with 33 kDa molecular mass. The protease activity had high stability when incubated at 10 or −20 °C for 6 months. The use of Metarhizium spores (1 × 104 spore mL−1) in combination with its protease (354.1 ± 2.2 U mL−1) completely inhibited hatching of the root-knot nematode Meloidogyne incognita within 24 h. Moreover, the mortality of juveniles increased to 100% at 48 h post treatment. The results were significantly different (p < 0.05) from Metarhizium spores or protease individually, as well as from extraction buffer and distilled water controls. Metarhizium spores suspended in crude protease rapidly germinated in 24 h and then mycelia invaded the immature eggs of M. incognita, while the eggs containing juveniles and the hatched juveniles were resistant. SEM observations showed extensive damage to eggshell and cuticle when treated with Metarhizium spore and protease, either individually or in combination. This is the first report on biocontrol potential of M. guizhouense against M. incognita. The combination of Metarhizium spores and protease has enhanced potential to develop into a safe and reliable method for plant parasitic nematode control.

Field evaluation of Duddingtonia flagrans IAH 1297 for the reduction of worm burden in grazing animals: Tracer studies in sheep

The aim of these studies was to determine the reduction in pasture infectivity likely to be achieved by the supplementation of grazing sheep with BioWorma®, a product containing the chlamydospores of the nematophagous fungus Duddingtonia flagrans strain IAH 1297. Four placebo-controlled trials were conducted between 2009 and 2013 in sheep in different climatic regions of New South Wales and Queensland, Australia and across several seasons. The effectiveness of BioWorma was assessed by total worm counts in tracer sheep placed in paddocks grazed by parasitised sheep which were fed a daily supplement with and without BioWorma under group-feeding conditions. Further proof of concept was obtained by assessing the worm burdens and weight gains of the parasitised sheep, as well as the number of anthelmintic (“salvage”) treatments required when faecal egg counts exceeded a threshold level.Significant reductions ranging from 57 to 84% (P < 0.05) in worm burdens of the tracer sheep placed in the paddock grazed by BioWorma treated sheep were obtained in all four trials, compared to the Control group. In two of the studies the treatment effect was greater at the end of the trial, indicating that pasture infectivity in the Control paddocks had risen considerably. The main nematodes encountered were Haemonchus spp., Trichostrongylus spp., and Teladorsagia spp. (including multi-resistant strains) and significant reductions were demonstrated for each of these species.Given the results of the four trials it can be concluded that supplementation of pastured sheep with BioWorma was effective in reducing the numbers of parasitic nematode larvae ingested by tracer sheep. It is considered that these levels of reduced pasture larvae would result in productivity increases in grazing sheep and reduce the requirement for intervention with anthelmintic chemicals. Therefore, use of BioWorma will provide an alternative means for control of gastrointestinal nematode (GIN) parasites on pasture.

Response of drug-susceptible and -resistant Haemonchus contortus larvae to monepantel and abamectin alone or in combination in vitro

There is an increasing interest in the use of combination anthelmintic products for the control of intestinal nematode parasites of livestock. These products are seen as attractive options for parasite control in the face of increasing levels of resistance to the different anthelmintic drug classes, as well as a means to slow the rate at which resistance develops to the individual components of the combination. With the recent introduction of an anthelmintic combination product containing abamectin and monepantel (at 1:12.5), we were interested in measuring the response of drug-susceptible and drug-resistant isolates of Haemonchus contortus to these two drugs alone and in combination, using larval development assays. The GWBII isolate showed resistance to abamectin (12-fold) alongside susceptibility to monepantel. The resistance ratio was reduced from 12- to 3.2-fold when the two drugs were combined. The MPL-R isolate was resistant to both drugs, with resistance factors of 6-fold towards abamectin, and 10.6- and 1008-fold towards monepantel in two sub-populations present in the isolate. This isolate showed 6.4-fold resistance to the drug combination. Hence, for both GWBII and MPL-R, the level of resistance towards the combination was reduced compared to the resistance towards abamectin or monepantel alone, respectively, but was not abolished. However, for GWBII, this in vitro resistance to the drug combination would be expected to have no impact on the in vivo efficacy of the combination drench product as the isolate is resistant to only the abamectin component of the drench, with monepantel remaining effective. On the other hand, the observed in vitro resistance to the combination shown by the MPL-R isolate is derived from significant levels of resistance towards both components separately, and hence may impact on in vivo efficacy of the combination. Isobologram analysis did not find any evidence for a synergistic interaction between the two drugs in larval development assays. We examined the predicted effects of varying the abamectin:monepantel ratio in drug combinations, assuming that the two drugs acted in an additive fashion. For GWBII, resistance to the drug combination was reduced to almost zero as the abamectin:monepantel ratio increased from 1:12.5 to 1:100, reflecting its resistance to only the abamectin component of the combination. For MPL-R, on the other hand, the resistance increased as the relative proportion of monepantel in the combination was increased, reflecting the extreme level of in vitro resistance shown by this isolate to monepantel.

In Vitro, in Vivo, and Interaction Studies of Nematophagous Fungus Arthrobotrys thaumasia ( Monacrosporium thaumasium) with the Larvae of Trichostrongylides of Sheep.

It is important to isolate potential candidates from the local isolates of nematophagous fungi and to investigate interaction between the fungal strains and gastrointestinal nematodes for the biological control of parasitic nematodes in livestock. In the present study, we assessed the in vitro predatory activity and the viability of isolates of Arthrobotrys thaumasia ( Monacrosporium thaumasium) after passage through the gastrointestinal tract of sheep. The predatory process of a representative isolate selected against the larvae of trichostrongylids was prepared with a scanning electron microscope (SEM). In vitro experiments tested the ability of 9 native isolates of A. thaumasia to prey on larvae of feces of sheep infected with natural mixed nematodes ( Haemonchus contortus, Trichostongylus colubriformis, Marshallagia mongolica). These isolates of A. thaumasia decreased infectivity of third stage infective larvae (L3) by 75.54-99.97%; 7 isolates decreased infectivity by more than 90%. In vivo experiments also demonstrated significant reductions of L3 numbers in the feces treated with the 9 isolates after passing through the gastrointestinal tract of sheep, and these decreases ranged from 51.68 to 88.16%. The isolates tested were re-isolated in 5-g sub-samples of feces from sheep in each treatment group, indicating that these isolates had the capacity to prey upon larvae of trichostrongylids after the passage through gastrointestinal tract. SEM shows that at 6 hr after the larvae were added, including the second stage larvae (L2) and L3 of trichostrongylids, the isolate NBS 005 caught them; at 8 hr after being caught L2 was penetrated by the fungus while penetration of L3 occurred at 12 hr; at 78 hr post-capture L2 was completely destroyed by the fungus while complete digestion of L3 occurred at 84 hr.

A BRIEF REVIEW ON THE MODE OF ACTION OF ANTINEMATODAL DRUGS.

Anthelmintics are some of the most widely used drugs in veterinary medicine. Here we review the mechanism of action of these compounds on nematode parasites. Included are the older classes of compounds; the benzimidazoles, cholinergic agonists and macrocyclic lactones. We also consider newer anthelmintics, including emodepside, derquantel and tribendimidine. In the absence of vaccines for most parasite species, control of nematode parasites will continue to rely on anthelmintic drugs. As a consequence, vigilance in detecting drug resistance in parasite populations is required. Since resistance development appears almost inevitable, there is a continued and pressing need to fully understand the mode of action of these compounds. It is also necessary to identify new drug targets and drugs for the continued effective control of nematode parasites.

Prevalence and Risk Factors of Gastrointestinal Nematodes Infections in Small Ruminants in Tullo District, Western Harerghe, Ethiopia

A cross-sectional study was carried out in sheep and goats kept under extensive management system in Tullo district, Western Harerghe zone of Oromia Regional State, Ethiopia, during the period from November, 2013 to April, 2014. The main objectives of the study were to estimate the prevalence of nematode infection in small ruminants and to assess associated risk factors with its occurrence. A total of 384 fecal samples were collected from 168 sheep and 216 goats. The samples were examined using floatation technique and eggs per gram of faeces were determined. The overall prevalence for the nematode infection was 50.8% (195/384). Different nematode eggs including Strongyle (177, 46.1%), Strongyloides (35, 9.1%) and Trichuris eggs (31, 8.1%) were identified. The mean eggs per gram count showed that, 100 (56.5%) were heavily infected, 61(34.5%) were moderately infected and the remaining were lightly infected. The coproculture examination from the positive samples for strongyle eggs revealed the presence of Trichostrongylus species (sp), Haemonchus sp., Oesophagostomum sp., Cooperia sp., and Bunostomum sp. Gastrointestinal nematode parasitism is more prevalent (AOR=2.23) in sheep as compared to goats. It was also higher in animals with poor (AOR=4.54) and medium body conditions (AOR=2.02) as compared to animals with good body conditions. However, there was no statistically significant association between the occurrence of the infection and age, sex and origin of the animals’. In conclusion, gastrointestinal nematode parasites were highly prevalent in the study area and this could hamper health and productivity of the small ruminants. Thus, strategic control of nematode parasites should be planned and implemented to reduce the economic losses and welfare problems of the animals.

In vitro control of parasitic nematodes of small ruminants using some plant species containing flavonoids.

This study determined in vitro anthelmintic efficacy of three plant species: Trema orientalis, Urtica dioica and Zanthozylum capense on nematode larvae of small ruminants. Dried leaf samples (40g) were extracted in 70% ethanol, in portions of 10g and concentrated to 100ml. Half and one quarter of the original crude extract were both made to 100ml. Rectal faecal material from 10 Merino sheep and 25 Nguni goats was pooled within species and thoroughly hand-mixed. Dung samples, each of 5g were cultured for 12days at 27°C. On day 13, 4 plates were watered and 4 others treated with ethanol to correct for solvent effect on mortality. The design was 2 (animal species) × 3 (plant species) × 3 (extract concentrations). In each of three runs, three plates were treated with each crude extract in three incremental concentrations. Surviving L3 larvae were isolated, counted and mortalities became indices of anthelmintic efficacy. Data from nematode larval mortality were analysed to determine the effect of animal species, plant species, concentration and their interactions. Efficacy was affected by concentration (P = 0.0001), animal species (P = 0.0046), plant species (P = 0.0572), the interactions of animal species and concentration (P = 0.0010), plant species and concentration (P = 0.0123) and concentration × animal × plant species (P = 0.0435).

In vitro and in vivo studies of the native isolates of nematophagous fungi from China against the larvae of trichostrongylides.

To screen potential nematophagous fungi candidates for the biological control of parasitic nematodes in livestock, in vitro and in vivo studies of the native isolates of nematophagous fungi against the larvae of trichostrongylides were conducted. The in vitro predatory activity of 16 native nematophagous fungal isolates on the larvae of trichostrongylides in sheep feces was assessed. In the ten isolates of Duddingtonia flagrans, the reduction percentage for the infective larvae (L3) of Trichostrongylus colubriformis ranged from 57.21 to 99.83%, and that of Haemonchus contortus ranged from 62.12 to 99.88%. The analysis of the same assay on five isolates of Arthrobotrys superba and one isolate of A. cookedickinson (Monacrosporium cystosporum) showed comparable results with those for D. flagrans. To determine the excretion time of fungal isolates in feces after oral administration, D. flagrans (SDH035) were studied in vivo in sheep and rabbits. Results showed that the tested fungal isolates existed in sheep feces from 12 to 72 h after fungal treatment, and the fungal excretion in rabbit feces occurred at 4 h, reached a peak at 10 h, and declined gradually 18 h after oral administration. All the native fungal isolates were assessed after passing through the gastrointestinal tract of sheep. Treatment with isolates of D. flagrans significantly reduced the number of developing larvae in the feces, and the efficacies ranged from 55.15 to 98.82%. One out of the five isolates of A. superba and A. cookedickinson (BS002) survived after passing through the gastrointestinal tract, and the L3 reduction rates were 83.79 and 81.33%, respectively. Results of the present study provide information about the in vitro predatory activity of nematophagous fungi from China on the L3 of trichostrongylides and their ability to pass through the gastrointestinal tract before administering them for biocontrol.

Abomasal mucosal immune responses of cattle with limited or continuous exposure to pasture-borne gastrointestinal nematode parasite infection

It has been well documented that cattle raised on pasture are slow in weight gain when compared to those fed with grain. Inflammation in the digestive system commonly caused by pasture-transmitted gastrointestinal (GI) nematode parasites that could negatively impact feed conversion has never been compared in cattle raised with no pasture exposure (NPE, uninfected), limited pasture exposure (LPE, exposure until weaning), or continuous pasture exposure (CPE, life time exposure). In the present study, the abomasal mucosal immune responses and inflammation of LPE and CPE cattle were investigated. Our results indicate that CPE cattle displayed inflamed abomasa with enlarged draining lymph nodes, the presence of Ostertagia ostertagi larvae and higher levels of Ostertagia-specific antibodies in circulation. The level of B cells was elevated in the abomasal mucosa in the presence (nodular) or absence (non-nodular) of Ostertagia-specific pathology, where B cells were 4-fold higher in the nodular mucosa. Foxp3+ CD4T cells were also noticeably elevated in both the abomasal mucosa and blood, but were only slightly higher in non-nodular mucosa than in the nodular mucosa of CPE animals. In contrast, LPE animals presented no enlargement of abomasal draining lymph nodes and exhibited little to no immune cell infiltration in the abomasal mucosa. Further, CPE animals had higher numbers of mucosal mast cells when compared to LPE animals, though mucosal mast cells were high in all animals. Overall, CPE cattle displayed significantly higher levels of inflammation and pathology in their abomasa and may explain in part slowed weight gain relative to LPE animals. The results of this study emphasize the need for GI nematode parasite control in CPE animals and development and application of vaccines which are compatible with the organic cattle production system.

Parasitic Nematode Immunomodulatory Strategies: Recent Advances and Perspectives

More than half of the described species of the phylum Nematoda are considered parasitic, making them one of the most successful groups of parasites. Nematodes are capable of inhabiting a wide variety of niches. A vast array of vertebrate animals, insects, and plants are all identified as potential hosts for nematode parasitization. To invade these hosts successfully, parasitic nematodes must be able to protect themselves from the efficiency and potency of the host immune system. Innate immunity comprises the first wave of the host immune response, and in vertebrate animals it leads to the induction of the adaptive immune response. Nematodes have evolved elegant strategies that allow them to evade, suppress, or modulate host immune responses in order to persist and spread in the host. Nematode immunomodulation involves the secretion of molecules that are capable of suppressing various aspects of the host immune response in order to promote nematode invasion. Immunomodulatory mechanisms can be identified in parasitic nematodes infecting insects, plants, and mammals and vary greatly in the specific tactics by which the parasites modify the host immune response. Nematode-derived immunomodulatory effects have also been shown to affect, negatively or positively, the outcome of some concurrent diseases suffered by the host. Understanding nematode immunomodulatory actions will potentially reveal novel targets that will in turn lead to the development of effective means for the control of destructive nematode parasites.

First report of the in vitro nematicidal effects of camel milk

Antipathogenic properties of camel milk have been investigated to substitute for drugs hence overcome drug resistance. The main objective of this present study was to investigate the anthelmintic activity of camel milk in relationship to its chemical composition. In vitro anthelmintic effects of camel milk against Haemonchus contortus from sheep were ascertained by egg hatching and worm motility inhibitions in comparison to milks from cow, ewe and goat as well as a reference drug albendazole. Chemical composition revealed that camel milk has higher contents of protective protein (lactoferrin) and vitamin C than other species’ milk. It showed ovicidal activity at all tested concentrations and completely inhibited egg hatching at a concentration close to 100mg/mL (inhibitory concentration (IC50)=42.39mg/mL). Camel milk revealed in vitro activity against adult parasites in terms of the paralysis and/or death of the worms at different hours post treatment. After 8h of exposure, it induced 100% mortality at the highest tested concentration. There was 82.3% immobility of worms in albendazole 8h post-exposition. No such effects were seen with the other species’ milks. Bioactive compounds such as lactoferrin and vitamin C may be involved in such an effect.To our knowledge, these results depict for the first time that camel milk possesses in vitro anthelmintic properties and further in vitro and in vivo trials against different parasite species and stages are required to make use of this milk for the control of gastrointestinal nematode parasites.

TGF-β signaling regulates resistance to parasitic nematode infection in Drosophila melanogaster

Over the past decade important advances have been made in the field of innate immunity; however, our appreciation of the signaling pathways and molecules that participate in host immune responses to parasitic nematode infections lags behind that of responses to microbial challenges. Here we have examined the regulation and immune activity of Transforming Growth Factor-beta (TGF-β) signaling in the model host Drosophila melanogaster upon infection with the nematode parasites Heterorhabditis gerrardi and H. bacteriophora containing their mutualistic bacteria Photorhabdus. We have found that the genes encoding the Activin and Bone Morphogenic Protein (BMP) ligands Dawdle (Daw) and Decapentaplegic (Dpp) are transcriptionally induced in flies responding to infection with the nematode parasites, containing or lacking their associated bacteria. We also show that deficient Daw or Dpp regulates the survival of D. melanogaster adults to the pathogens, whereas inactivation of Daw reduces the persistence of the nematodes in the mutant flies. These findings demonstrate a novel role for the TGF-β signaling pathways in the host anti-nematode immune response. Understanding the molecular mechanisms of host anti-nematode processes will potentially lead to the development of novel means for the efficient control of parasitic nematodes.

Molecular mechanisms for anthelmintic resistance in strongyle nematode parasites of veterinary importance.

Veterinarians rely on a relatively limited spectrum of anthelmintic agents to control nematode parasites in domestic animals. Unfortunately, anthelmintic resistance has been an emerging problem in veterinary medicine. In particular, resistance has emerged among the strongyles, a group of gastrointestinal nematodes that infect a variety of hosts that range from large herbivores to small companion animals. Over the last several decades, a great deal of research effort has been directed toward developing an understanding of the mechanisms conferring resistance against the three major groups of anthelmintics: macrocyclic lactones, benzimidazoles, and nicotinic agonists. Our understanding of anthelmintic resistance has been largely formed by determining the mechanism of action for each drug class and then evaluating drug-resistant nematode isolates for mutations or differences in expression of target genes. More recently, drug efflux pumps have been recognized for their potential contribution to anthelmintic resistance. In this mini-review, we summarize the evidence for mechanisms of resistance in strongyle nematodes.

In vitro and in vivo anthelmintic activities of Iris kashmiriana Linn.

In search of a natural antiparasitic, in vitro and in vivo anthelmintic activities of methanol and aqueous extracts of Iris kashmiriana Linn. rhizome were tested against gastrointestinal nematodes of sheep. A worm motility inhibition assay was used for in vitro study and a faecal egg count reduction assay was used for in vivo study. Crude aqueous extracts of I. kashmiriana exhibited greater anthelmintic activity against Haemonchus contortus than crude methanolic extract (P<0.05). The aqueous extracts of I. kashmiriana resulted in a mean worm motility inhibition of 85.0%, while methanolic extracts resulted in a mean worm motility inhibition of 100.0%. The in vivo anthelmintic activity of aqueous and methanolic extracts of I. kashmiriana in sheep naturally infected with mixed gastrointestinal nematodes species demonstrated a maximum (70.27%) egg count reduction in sheep treated with aqueous extracts at 2gkg−1 body weight on day 15 after treatment, closely followed by methanolic extracts at 1gkg−1 body weight on day 15 after treatment (33.17% egg count reduction) while as lethal concentration (LC50) values of aqueous and methanolic extracts of I. kashmiriana on adult worms of gastrointestinal nematodes of sheep H. contortus are 18.50mg/ml and 16.66mg/ml respectively. Thus aqueous extracts exhibited greater anthelmintic activity under both in vitro and in vivo conditions; this could be due to the presence of water soluble active ingredients in I. kashmiriana. From the present study it can be suggested that I. kashmiriana rhizome exhibited significant anthelmintic activity against gastrointestinal nematodes of sheep and has the potential to contribute to the control of gastrointestinal nematode parasites of small ruminants.

Season-long effectiveness of stocker-calf treatment at turnout with eprinomectin extended-release injection or a combination of injectable doramectin and oral albendazole

On May 22, 2013, 128 stocker calves enrolled in an internal parasite control study were treated according to structured allocations and placed directly onto treatment-specific, randomly allocated, similar pastures (4 animals/2 acres (0.8 hectare) pasture). Treatment groups were saline injection (8 pastures); 0.09 mg/lb (0.2 mg/kg) BW doramectin injection concomitantly with 4.54 mg/lb (10 mg/kg) BW albendazole oral suspension (12 pastures); and 0.45 mg/lb (1 mg/kg) BW eprinomectin extended-release injection (12 pastures). Over the 119-day grazing period, average daily gains ñ SE were 1.21ñ0.07, 1.46ñ0.04, and 1.52ñ0.04 lb (0.55ñ0.03, 0.66ñ0.02, and 0.69ñ0.02 kg)/day for the saline, combination, and extended-release groups, respectively; calves in the combination and extended-release groups outgained the control group (P&lt;0.05). In comparison to the saline-treated group, there were statistically significant (P&lt;0.05) reductions of strongyle fecal egg counts at 14, 30, and 58 days post-treatment (combination group) and at 14 and 30 days post-treatment (extended-release group). No significant differences in adult Haemonchus placei, Ostertagia ostertagi, Cooperia punctata, and Oesophagostomum radiatum populations between treatment groups were seen at necropsy. The combination treatment group had fewer O. ostertagi early fourth-stage larvae than did controls (P&lt;0.05). Based on results of this study, a single treatment of heavily infected and challenged stocker calves in the spring in Arkansas with either extended-release eprinomectin or doramectin+albendazole was not sufficient for adequate parasitic nematode control.

Determinación del crecimiento de hongos nematófagos en diversas fuentes de carbono

Las enmiendas orgánicas han sido utilizadas para estimular la acción antagónica de los hongos nematófagos depredadores (HND) del suelo, sin embargo, su uso ha producido resultados inconsistentes en el control de nematodos parásitos. Las inconsistencias han sido atribuidas parcialmente a la composición de las enmiendas orgánicas, específicamente al contenido de carbono y nitrógeno. Conocer las preferencias de fuentes de carbono podría ser de utilidad para favorecer la fase depredadora de los HND del suelo. El presente trabajo tiene por objetivo determinar el crecimiento de HND nativos de Costa Rica en fuentes de carbono diversas. Para esto, cepas de HND de las especies Candelabrella musiformis y Arthrobotrys oligospora fueron cultivadas en medios artificiales con diversas fuentes de carbono. La velocidad de crecimiento desarrollada por los HND en los medios de cultivo fue determinada y comparada. Las cepas de C. musiformis y de A. oligospora desarrollaron velocidades de crecimiento en las fuentes de carbono en el siguiente orden: celulosa&gt;quitina&gt; pectina&gt;almidón&gt;leche descremada. Únicamente hubo diferencias significativas en la velocidad de crecimiento en el medio con celulosa, en comparación con los otros medios de cultivo. En dicho medio ambas especies fúngicas crecieron más rápidamente, pero las cepas de A. oligospora crecieron más rápidamente en comparación con las cepas de C. musiformis. Ambas especies de HND desarrollaron velocidades de crecimiento bajas en los medios que contenían almidón y leche descremada.