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The antibiotic and nematocidal activities of extracts from two coastal lichen species collected on Lampedusa Island (Sicily), Ramalina implexa Nyl. and Roccella phycopsis Ach., were tested. Methyl orsellinate, orcinol, (+)-montagnetol, and for the first time 4-chlororcinol were isolated from Roccella phycopsis. (+)-Usnic acid was obtained from Ramalina implexa. The crude organic extract of both lichen species showed strong antibiotic activity against some bacterial species and nematocidal activity. Among all the pure metabolites tested against the infective juveniles (J2) of the root-knot nematode (RKN) Meloydogine incognita, (+)-usnic acid, orcinol, and (+)-montagnetol had significant nematocidal activity, comparable with that of the commercial nematocide Velum® Prime, and thus they showed potential application in agriculture as a biopesticide. On the contrary, methyl orsellinate and 4-chlororcinol had no nematocidal effect. These results suggest that the substituent pattern at ortho-para-position in respect to both hydroxyl groups of resorcine moiety, which is present in all metabolites, seems very important for nematocidal activity. The organic extracts of both lichens were also tested against some Gram-positive and Gram-negative bacteria. Both extracts were active against Gram-positive species. The extract of Ramalina implexa showed, among Gram-negative species, activity against Escherichia coli and Acinetobacter baumannii, while that from Roccella phycopsis was effective towards all test strains, with the exception of Pseudomonas aeruginosa. The antimicrobial activity of (+)-usnic acid, methyl orsellinate, and (+)-montagnetol is already known, so tests were focused on orcinol and 4-chlororcinol. The former showed antibacterial activity against all Gram positive and Gram-negative test strains, with the exception of A. baumannii and K. pneumoniae, while the latter exhibited a potent antibacterial activity against Gram-positive test strains and among Gram-negative strains, was effective against A. baumannii and K. pneumonia. These results suggest, for orcinol and 4-chlororcinol, an interesting antibiotic potential against both Gram-positive and Gram-negative bacterial strains.

Root-knot nematodes (RKN) such as Meloidogyne spp. are among the most detrimental pests in agriculture affecting several crops. New methodologies to manage RKN are needed such as efficient discovery of nematophagous microbes. In this study, we developed an in vitro high-throughput method relying on the free-living nematode Caenorhabditis elegans and the infection of those nematodes with a soil slurry containing a microbiome likely to house nematophagous microbes. Nematodes were monitored for presence of infection and sub-cultured repeatedly for the purpose of isolating pure cultures of the microbe responsible for conferring the nematicidal activity. Once soil microbes were confirmed to be antagonistic to C. elegans, they were tested for pathogenicity against Meloidogyne chitwoodi. Using this methodology, the fungal isolate Mortierella globalpina was confirmed to be pathogenic in vitro against M. chitwoodi by nematode trapping via hyphal adhesion to the cuticle layer, penetration of the cuticle layer, and subsequently digestion of its cellular contents. M. globalpina was also observed to reduce disease symptomology of RKNs in vivo via significant reduction of root-galls on tomato (Solanum lycopersicum var. Rutgers).

The discovery of novel drugs against animal parasites is in high demand due to drug-resistance problems encountered around the world. Herein, the synthesis and characterization of 27 organic and organometallic derivatives of the recently launched nematocidal drug monepantel (Zolvix®) are described. The compounds were isolated as racemates and were characterized by 1H, 13C, and 19F NMR spectroscopy, mass spectrometry, and IR spectroscopy, and their purity was verified by microanalysis. The molecular structures of nine compounds were confirmed by X-ray crystallography. The anthelmintic activity of the newly designed analogues was evaluated in vitro against the economically important parasites Haemonchus contortus and Trichostrongylus colubriformis. Moderate nematocidal activity was observed for nine of the 27 compounds. Three compounds were confirmed as potentiators of a known monepantel target, the ACR-23 ion channel. Production of reactive oxygen species may confer secondary activity to the organometallic analogues. Two compounds, namely, an organic precursor (3 a) and a cymantrene analogue (9 a), showed activities against microfilariae of Dirofilaria immitis in the low microgram per milliliter range.

The essential phosphobase methylation pathway for synthesis of phosphocholine is unique to nematodes, protozoa and plants, and thus an attractive antiparasitic molecular target. Herein, we screened compounds from the National Cancer Institute (Developmental Therapeutics Program Open Chemical Repository) for specific inhibitory activity against Haemonchus contortus phosphoethanolamine methyltransferases (HcPMT1 and HcPMT2), and tested candidate compounds for anthelmintic activity against adult and third-stage larvae of H. contortus. We identified compound NSC-641296 with IC50 values of 8.3 ± 1.1 μM and 5.1 ± 1.8 μM for inhibition of the catalytic activity of HcPMT1 alone and HcPMT1/HcPMT2 combination, respectively. Additionally we identified compound NSC-668394 with inhibitory IC50 values of 5.9 ± 0.9 μM and 2.8 ± 0.6 μM for HcPMT1 alone and HcPMT1/HcPMT2 combination, respectively. Of the two compounds, NSC-641296 depicted significant anthelmintic activity against third-stage larvae (IC50 = 15 ± 2.9 μM) and adult stages (IC50 = 7 ± 2.9 μM) of H. contortus, with optimal effective in vitro concentrations being 2-fold and 4-fold, respectively, lower than its cytotoxic IC50 (29 ± 2.1 μM) in a mammalian cell line. Additionally, we identified two compounds, NSC-158011 and NSC-323241, with low inhibitory activity against the combined activity of HcPMT1 and HcPMT2, but both compounds did not show any anthelmintic activity against H. contortus. The identification of NSC-641296 that specifically inhibits a unique biosynthetic pathway in H. contortus and has anthelmintic activity against both larval and adult stages of H. contortus, provides impetus for the development of urgently needed new efficacious anthelmintics to address the prevailing problem of anthelmintic-resistant H. contortus., Graphical abstract, Highlights • NSC-641296 and NSC-668394 inhibit HcPMT1 and HcPMT2 enzymes in Haemonchus contortus. • NSC-641296 has in vitro anthelmintic activity against larvae and adult H. contortus. • H. contortus HcPMT1 and HcPMT2 are two unique targets for anthelmintic development.

Schistosomiasis is a geographically restricted disease caused by several species of the trematode worm Schistosoma. If untreated, it can lead to major disability and death. Typically, only nonspecific symptoms, such as cachexia and anemia occur, but occasionally patients experience more severe complications arising from hepatic (portal hypertension and its related consequences) and urinary involvement. Although in the past few years, preventive measures have reduced the worldwide burden of infection, active transmission still occurs on several different continents...

The control of parasitic roundworms (nematodes) is heavily reliant on the use of a limited number of anthelmintic drugs. However, drug resistance is now very widespread and no vaccines are available, such that the discovery of new chemical entities is crucial. Within this context, we screened a library of pure natural products (n=400) against exsheathed third-stage (xL3) larvae of the parasitic nematode Haemonchus contortus using a whole-organism screening method. We identified two plant-derived rotenoids, deguelin and rotenone, with inhibitory activity on xL3 motility. Rotenone was not investigated further, because of its toxicity to some vertebrates. The dose response and cytotoxicity studies showed potent and selective inhibitory activity of deguelin on motility of xL3 larvae of H. contortus. Detailed future work needs to be conducted to explore the mode of action of this compound on H. contortus and related nematodes, and to assess its potential as an anthelmintic candidate.

The L3-stage-specific chitinase OvCHT1 has been implicated in the development of Onchocerca volvulus, the causative agent of onchocerciasis. Closantel, a known anthelmintic drug, was previously discovered as a potent and specific OvCHT1 inhibitor. As closantel is also a known protonophore, we performed a simple scaffold modulation to map out the structural features that are relevant for its individual or dual biochemical roles. Furthermore, we present that either OvCHT1 inhibition or protonophoric activity was capable of affecting O. volvulus L3 molting and that the presence of both activities in a single molecule yielded more potent inhibition of the nematode’s developmental process.

The completion of a number of nematode genomes has provided significant information on ABC systems in these organisms. Nematodes have more ABC systems genes and greater diversity than do mammalian species. Class 1 and class 2 ABC systems, more commonly known as ABC transporters, are present. As in other organisms, nematode ABC systems are characterized by a highly conserved ATP-binding domain (ABC_2) and a less conserved transmembrane domain (ABC_TM1/TM1F). Studies of drug resistance in nematodes have suggested that ABC transporters are part of the resistance mechanism. Evidence in support of this has been obtained from genetic studies where an association between anthelmintic selection and ABC transporters was shown by comparisons between unselected and drug selected, or resistant, populations of parasitic nematodes. In drug resistant populations, genetic polymorphism and diversity, genotype patterns, and linkage disequilibrium were disrupted. Multidrug resistance (MDR) reversing agents that inhibit ABC function improve efficacy in sensitive nematode populations and restore sensitivity in resistant populations. Similar to the situation in clinical oncology, overexpression of ABC systems occurs in drug resistant and sensitive populations following drug exposure, particularly those in the P-glycoprotein (PGP) subfamily. Deletion or disruption of ABC genes, particularly PGP and the multidrug resistance associated protein (MRP), increases sensitivity to some drugs, particularly ivermectin. These studies provide evidence that ABC transporters play a role in drug action and resistance in nematodes.

SUMMARYPlant cysteine proteinases (CPs) from papaya (Carica papaya) are capable of killing parasitic nematode wormsin vitroand have been shown to possess anthelmintic effectsin vivo.The acute damage reported in gastrointestinal parasites has not been found in free-living nematodes such asCaenorhabditis elegansnor among the free-living stages of parasitic nematodes. This apparent difference in susceptibility might be the result of active production of cysteine proteinase inhibitors (such as cystatins) by the free-living stages or species. To test this possibility, a supernatant extract of refined papaya latex (PLS) with known active enzyme content was used. The effect on wild-type (Bristol N2) and cystatin null mutant (cpi-1−/−andcpi-2−/−)C. eleganswas concentration-, temperature- and time-dependent. Cysteine proteinases digested the worm cuticle leading to release of internal structures and consequent death. Both cystatin null mutant strains were highly susceptible to PLS attack irrespective of the temperature and concentration of exposure, whereas wild-type N2 worms were generally resistant but far more susceptible to attack at low temperatures. PLS was able to induce elevatedcpi-1 andcpi-2 cystatin expression. We conclude that wild-typeC. elegansdeploy cystatins CPI-1 and CPI-2 to resist CP attack. The results suggest that thecpi-1 orcpi-2 null mutants (or a double mutant combination of the two) could provide a cheap and effective rapid throughputC. elegans-based assay for screening plant CP extracts for anthelmintic activity.

Two new tenvermectins, tenvermectins C (1) and D (2), were isolated from Streptomyces avermitilis MHJ1011. Their structures were determined by extensive spectroscopic analysis and by comparison with a related known compound, tenvermectin A. Compounds 1 and 2 exhibited potent nematocidal and acaricidal activities against Bursaphelenchus xylophilus and Tetranychus cinnabarinus.

This study utilised computer simulation modelling (Risk Management Model for Nematodes) to investigate the impact of different parasite refugia scenarios on the development of anthelmintic resistance and worm control effectiveness. The simulations were conducted for adult ewe flocks in a Mediterranean climatic region over a 20 year time period. Factors explored in the simulation exercise were environment (different weather conditions), drug efficacy, the percentage of the flock left untreated, the timing of anthelmintic treatments, the initial worm egg count, and the number of drenches per annum. The model was run with variable proportions of the flock untreated (0, 10, 20, 30, 40 and 50%), with ewes selected at random so that reductions in the mean worm burden or egg count were proportional to the treated section of the flock. Treatments to ewes were given either in summer (December; low refugia potential, hence highly selective) or autumn (March; less selective due to a greater refugia potential), and the use of different anthelmintics was simulated to indicate the difference between active ingredients of different efficacy. Each model scenario was run for two environments, specifically a lower rainfall area (more selective) and a higher rainfall area (less selective) within a Mediterranean climatic zone, characterised by hot, dry summers and cool, wet winters. Univariate general linear models with least square difference post-hoc tests were used to examine differences between means of factors. The results confirmed that leaving a proportion of sheep in a flock untreated was effective in delaying the development of anthelmintic resistance, with as low as 10% of a flock untreated sufficient to significantly delay resistance, although this strategy was associated with a small reduction in worm control. Administering anthelmintics in autumn rather than summer was also effective in delaying the development of anthelmintic resistance in the lower rainfall environment where all sheep were treated, although the effect of treatment timing on worm control effectiveness varied between the environments and the proportion of ewes left untreated. The use of anthelmintics with higher efficacy delayed the development of resistance, but the initial worm egg count or number of annual treatments had no effect on either the time to resistance development or worm control effectiveness. In conclusion, the modelling study suggests that leaving a small proportion of ewes untreated, or changing the time of treatment, can delay the onset of anthelmintic resistance in a highly selective environment.

This review discusses the latest approaches to the diagnosis and treatment of patients with strongyloidiasis, with an emphasis on infection in the immunocompromised host and the risk for disseminated strongyloidiasis.The differences in acute, chronic, accelerated autoinfection, and disseminated disease in Strongyloides stercoralis infection are explored with particular emphasis on early diagnosis, treatment, and prevention. The goals of treatment are investigated for the different infection states. Predisposing risks for dissemination are delineated, and the roles played for newer diagnostics in the identification of at-risk individuals are detailed.The use of newer diagnostic tests and broader screening of immunocompromised patients from Strongyloides-endemic areas is of paramount importance, particularly if prevention of life-threatening dissemination is the goal.

Current control practices against gastrointestinal nematodes in dairy cows rely strongly on anthelmintic use. To reduce the development of anthelmintic resistance or disposition of drug residues in the environment, novel control approaches are currently proposed that target anthelmintic treatment to individual animals instead of the whole herd. However, such selective treatment strategies come with additional costs for labor and diagnostics and, so far, no studies have addressed whether they could be economically sustainable. The objectives of this study were to (1) investigate the economic effects at farm level of whole-herd versus more selective anthelmintic treatment strategies in adult dairy cows, and (2) determine how these economic effects depend on level of infection and herd size. A Monte Carlo simulation, fed by current epidemiological and economical knowledge, was used to estimate the expected economic effects and possible variation of different control strategies under Belgian conditions. Four treatment strategies were compared with a baseline situation in which no treatments were applied: whole herd at calving (S1), selective at calving with (S2) or without (S3) treatment of the first-calf cows, and wholeherd when animals are moved from grazing to the barn in the fall (housing treatment, S4). The benefit per lactation for an average dairy herd varied between -$2 and $131 (average $64) for S1, between -$2 and $127 (average $62) for S2, between -$17 and $104 (average $43) for S3, and between -$41 and $72 (average $15) for S4. The farmer’s risk associated with any treatment strategy, as indicated by the width of the 95% credible intervals of economic benefit of anthelmintic treatment, decreased with increasing level of exposure, as assessed by bulk tank milk ELISA. The order of the different strategies when sorted by expected benefit was robust to changes in economic input parameters. We conclude that, on average, strategies applying anthelmintic treatment at calving outperform a strategy applying treatment at housing. Within the strategies that applied treatment at calving, more selective treatment strategies can be economically sustainable. However, given the large variation in possible benefits within each treatment strategy, decision support systems are needed to account for the multitude of cow, epidemiological, and economic factors that determine the economics of nematode control and select the optimal treatment strategy for a specific farm. Key words: dairy , nematode , anthelmintic , economic

BACKGROUND AND PURPOSE: The cholinergic agonist levamisole is widely used to treat parasitic nematode infestations. This anthelmintic drug paralyses worms by activating a class of levamisole-sensitive acetylcholine receptors (L-AChRs) expressed in nematode muscle cells. However, levamisole efficacy has been compromised by the emergence of drug-resistant parasites, especially in gastrointestinal nematodes such as Haemonchus contortus. We report here the first functional reconstitution and pharmacological characterization of H. contortus L-AChRs in a heterologous expression system. EXPERIMENTAL APPROACH: In the free-living nematode Caenorhabditis elegans, five AChR subunit and three ancillary protein genes are necessary in vivo and in vitro to synthesize L-AChRs. We have cloned the H. contortus orthologues of these genes and expressed them in Xenopus oocytes. We reconstituted two types of H. contortus L-AChRs with distinct pharmacologies by combining different receptor subunits. KEY RESULTS: The Hco-ACR-8 subunit plays a pivotal role in selective sensitivity to levamisole. As observed with C. elegans L-AChRs, expression of H. contortus receptors requires the ancillary proteins Hco-RIC-3, Hco-UNC-50 and Hco-UNC-74. Using this experimental system, we demonstrated that a truncated Hco-UNC-63 L-AChR subunit, which was specifically detected in a levamisole-resistant H. contortus isolate, but not in levamisole-sensitive strains, hampers the normal function of L-AChRs, when co-expressed with its full-length counterpart. CONCLUSIONS AND IMPLICATIONS: We provide the first functional evidence for a putative molecular mechanism involved in levamisole resistance in any parasitic nematode. This expression system will provide a means to analyse molecular polymorphisms associated with drug resistance at the electrophysiological level.

Drug resistance in parasitic nematodes is an increasing problem worldwide, with resistance reported to all three commonly used classes of anthelmintics. Most studies to date have sought to correlate the resistant phenotype with genotypic changes in putative target molecules. Although this approach has identified mutations in several relevant genes, resistance might result from a complex interaction of different factors. Here we propose an alternative mechanism underlying the development of drug resistance based on functional differences in microRNA activity in resistant parasites. microRNAs play an important role in resistance to chemotherapeutic agents in many tumour cells and here we discuss whether they might also be involved in anthelmintic resistance in parasitic nematodes.

Caenorhabditis elegans, a free-living soil nematode, is an ideal model system for studying various physiological problems relevant to human diseases. Despite its short history, C. elegans proteomics is receiving great attention in multiple research areas, including the genome annotation, major signaling pathways (e.g. TGF-beta and insulin/IGF-1 signaling), verification of RNA interference-mediated gene targeting, aging, disease models, as well as peptidomic analysis of neuropeptides involved in behavior and locomotion. For example, a proteome-wide profiling of developmental and aging processes not only provides basic information necessary for constructing a molecular network, but also identifies important target proteins for chemical modulation. Although C. elegans has a simple body system and neural circuitry, it exhibits very complicated functions ranging from feeding to locomotion. Investigation of these functions through proteomic analysis of various C. elegans neuropeptides, some of which are not found in the predicted genome sequence, would open a new field of peptidomics. Given the importance of nematode infection in plants and mammalian pathogenesis pathways, proteomics could be applied to investigate the molecular mechanisms underlying plant- or animal-nematode pathogenesis and to identify novel antinematodal drugs. Thus, C. elegans proteomics, in combination of other molecular, biological and genetic techniques, would provide a versatile new tool box for the systematic analysis of gene functions throughout the entire life cycle of this nematode.

The antinematodal activity and mechanism of a 23-mer antimicrobial peptide, PMAP-23, derived from pig myeloid was investigated. PMAP-23 displayed a strong antinematodal activity against the eggs and worms of Caenorhabditis elegans. To investigate the antinematodal mechanism of PMAP-23, fluorescence activated flow cytometry and confocal laser scanning microscopy were performed. C. elegans treated with PMAP-23 showed higher fluorescence intensity by propidium iodide (PI) staining than normal cells. Confocal microscopy showed that the peptide was localized in the egg's shell and cell membrane. The action of the peptide against C. elegans membranes was examined by testing the membrane disrupting activity using liposome (PC/PS; 3:1, w/w). The result suggests that PMAP-23 may exert its antinematodal activity by disrupting the structure of the cell membrane via pore formation or via direct interaction with the lipid bilayers.

Successive extracts of Tribulus terrestris prepared using petroleum ether, chloroform, 50% methanol and water were tested for anthelmintic activity in-vitro using the nematode Caenorhabditis elegans. The activity could be detected only in 50% methanol extract which on further bioactivity guided fractionation and chromatographic separation yielded a spirostanol type saponin, tribulosin and beta-sitosterol-D-glucoside. Both the compounds exhibited anthelmintic activity with ED50 of 76.25 and 82.50 microg/ml respectively.

The present study proved that combined administration of pyrantel and febantel to Heterakis spumosa-infected mice yielded clear synergistic effects (seen in a quicker expulsion of the worms and a significant higher degree of worm degeneration), whereas the different doses of both drugs never reached the same efficacy, when given alone. It is concluded that the synergistic action of pyrantel and febantel on the functions of different organs of the parasites (muscle, nerve, inertine etc.) seen in the rodent model - also holds for the gut dwelling nematodes of the dog.

(1997). Bioreactor Cultivation of the Nematode Caenorhabditis elegans: Large Scale Production of Biologically Active Drug Receptors for Pharmaceutical Research. Biotechnology and Genetic Engineering Reviews: Vol. 14, No. 1, pp. 37-50.

Background The lack of new anthelmintic agents is of growing concern because it affects human health and our food supply, as both livestock and plants are affected. Two principal factors contribute to this problem. First, nematode resistance to anthelmintic drugs is increasing worldwide and second, many effective nematicides pose environmental hazards. In this paper we address this problem by deploying a high throughput screening platform for anthelmintic drug discovery using the nematode Caenorhabditis elegans as a surrogate for infectious nematodes. This method offers the possibility of identifying new anthelmintics in a cost-effective and timely manner. Methods/Principal findings Using our high throughput screening platform we have identified 14 new potential anthelmintics by screening more than 26,000 compounds from the Chembridge and Maybridge chemical libraries. Using phylogenetic profiling we identified a subset of the 14 compounds as potential anthelmintics based on the relative sensitivity of C. elegans when compared to yeast and mammalian cells in culture. We showed that a subset of these compounds might employ mechanisms distinct from currently used anthelmintics by testing diverse drug resistant strains of C. elegans. One of these newly identified compounds targets mitochondrial complex II, and we used structural analysis of the target to suggest how differential binding of this compound may account for its different effects in nematodes versus mammalian cells. Conclusions/Significance The challenge of anthelmintic drug discovery is exacerbated by several factors; including, 1) the biochemical similarity between host and parasite genomes, 2) the geographic location of parasitic nematodes and 3) the rapid development of resistance. Accordingly, an approach that can screen large compound collections rapidly is required. C. elegans as a surrogate parasite offers the ability to screen compounds rapidly and, equally importantly, with specificity, thus reducing the potential toxicity of these compounds to the host and the environment. We believe this approach will help to replenish the pipeline of potential nematicides., Author Summary With over two billion people infected and many billions of dollars of lost crops annually, nematode infections are a serious problem for human health and for agricultural production. While there are drugs to treat infections, many pockets of parasites have been identified worldwide that are developing immunity to the standard treatment regimen. In this study we describe a strategy using the model organism C. elegans as a surrogate parasite to identify several new chemical compounds that may offer additional treatments for infection. We demonstrate how to use our platform to identify compounds that are specific in their effect to nematodes and are not simply biocides. We also show through genetic and molecular analysis in this organism that we can quickly identify the mode of action of any new compound. Most critically, we show that a compound first identified in a free-living nematode, Caenorhabditis elegans, is also effective on a parasitic nematode, Meloidogyne hapla. With this result and considering the level of sequence conservation across much of the nematode phyla we believe our strategy can be more widely applied to find new anthelmintics.

Current control strategies for onchocerciasis and lymphatic filariasis (LF) rely on prolonged yearly or twice-yearly mass administration of microfilaricidal drugs. Prospects for near-term elimination or eradication of these diseases would be improved by availability of a macrofilaricide that is highly effective in a short regimen. Flubendazole (FLBZ), a benzimidazole anthelmintic registered for control of human gastrointestinal nematode infections, is a potential candidate for this role. FLBZ has profound and potent macrofilaricidal effects in many experimental animal models of filariases and in one human trial for onchocerciasis after parental administration. Unfortunately, the marketed formulation of FLBZ provides very limited oral bioavailability and parenteral administration is required for macrofilaricidal efficacy. A new formulation that provided sufficient oral bioavailability could advance FLBZ as an effective treatment for onchocerciasis and LF. Short-term in vitro culture experiments in adult filariae have shown that FLBZ damages tissues required for reproduction and survival at pharmacologically relevant concentrations. The current study characterized the long-term effects of FLBZ on adult Brugia malayi by maintaining parasites in jirds for up to eight weeks following brief drug exposure (6–24 hr) to pharmacologically relevant concentrations (100 nM—10 μM) in culture. Morphological damage following exposure to FLBZ was observed prominently in developing embryos and was accompanied by a decrease in microfilarial output at 4 weeks post-exposure. Although FLBZ exposure clearly damaged the parasites, exposed worms recovered and were viable 8 weeks after treatment., Author Summary Onchocerciasis and lymphatic filariasis are debilitating diseases caused by infections with filarial nematodes. The World Health Organization aims to eliminate these infections as public health problems. Despite prolonged control efforts, including chemotherapy through mass drug administration (MDA), transmission and infections persist. Addition of a microfilaricide that is efficacious in a short regimen would enhance prospects for achieving elimination goals. We investigated the long-term effects of the macrofilaricidal drug, flubendazole (FLBZ), on Brugia malayi. Adult parasites were exposed in culture to FLBZ at pharmacologically relevant concentrations (100 nM—10 μM) for up to 24 hr prior to implantation into the abdominal cavity of a jird for long-term maintenance. The greatest drug effect was on embryogenesis; morphological damage was most evident in early developmental stages. There was also a decrease in the release of microfilaria (mf) from the adult. Interestingly, no damage was observed to fully formed mf. Although further studies are required to determine to what extent these findings can be extrapolated to a field setting, an exposure profile which may produce similar effects in vivo has been defined.

The need for novel antibacterial strategies and the awareness of the importance of quorum sensing (QS) in bacterial infections have stimulated research aimed at identifying QS inhibitors (QSIs). However, clinical application of QSIs identified so far is still distant, likely due to their unsuitability for use in humans. A promising way to overcome this problem is searching for anti-QS side activity among the thousands of drugs approved for clinical use in the treatment of different diseases. Here, we applied this strategy to the search for QSIs, by screening a library of FDA-approved compounds for their ability to inhibit the QS response in the Gram-negative pathogen Pseudomonas aeruginosa . We found that the anthelmintic drug niclosamide strongly inhibits the P. aeruginosa QS response and production of acyl-homoserine lactone QS signal molecules. Microarray analysis showed that niclosamide affects the transcription of about 250 genes, with a high degree of target specificity toward the QS-dependent regulon. Phenotypic assays demonstrated that niclosamide suppresses surface motility and production of the secreted virulence factors elastase, pyocyanin, and rhamnolipids, and it reduces biofilm formation. In accordance with the strong antivirulence activity disclosed in vitro , niclosamide prevented P. aeruginosa pathogenicity in an insect model of acute infection. Besides the finding that an FDA-approved drug has a promising antivirulence activity against one of the most antibiotic-resistant bacterial pathogens, this work provides a proof of concept that a lateral anti-QS activity can be detected among drugs already used in humans, validating a new approach to identify QSIs that could easily move into clinical applications.

To the EditorEosinophilic myocarditis is a relatively rare condition and usually results from myocardial damage as a result of drugs or parasites. It is generally associated with increased peripheral eosinophil count.1 The clinical presentation of patients with myocarditis is variable, ranging from

This paper describes a new microfluidic platform for screening drugs and their dose response on the locomotion behavior of free living nematodes and parasitic nematodes. The system offers a higher sensitivity drug screening chip which employs a combination of existing and newly developed methods. Real-time observation of the entire drug application process (i.e. the innate pre-exposure locomotion, the transient response during drug exposure and the time-resolved, post-exposure behavior) at a single worm resolution is made possible. The chip enables the monitoring of four nematode parameters (number of worms responsive, number of worms leaving the drug well, average worm velocity and time until unresponsiveness). Each parameter generates an inherently different dose response; allowing for a higher resolution when screening for resistance. We expect this worm chip could be used as a robust cross species, cross drug platform. Existing nematode motility and migration assays do not offer this level of sophistication. The device comprises two principal components: behavioral microchannels to study nematode motility and a drug well for administering the dose and observing drug effects as a function of exposure time. The drug screening experiment can be described by three main steps: (i) ‘pre-exposure study’ – worms are inserted into the behavioral channels and their locomotion is characterized, (ii) ‘dose exposure’ – worms are guided from the behavioral microchannels into the drug well and held for a predefined time, during which time their transient response to the dose is characterized and (iii) ‘post-exposure study’ – worms are guided back into the behavioral microchannels where their locomotion (i.e. their time-resolved response to the dose) is characterized and compared to pre-exposure motility. The direction of nematodes' movement is reliably controlled by the application of an electric field within a defined range. Control experiments (e.g. in the absence of any drug) confirm that the applied electric fields do not affect the worms' motility or viability. We demonstrate the workability of the microfluidic platform on free living Caenorhabditis elegans (wild-type N2 and levamisole resistant ZZ15 lev-8) and parasitic Oesophagotomum dentatum (levamisole-sensitive, SENS and levamisole-resistant, LEVR) using levamisole (a well-studied anthelmintic) as the test drug. The proposed scheme of drug screening on a microfluidic device is expected to significantly improve the resolution, sensitivity and data throughput of in vivo testing, while offering new details on the transient and time-resolved exposure effects of new and existing anthelmintics.

The regulatory constraints imposed on use of chemical control agents in agriculture are rendering crops increasingly vulnerable to plant parasitic nematodes. Thus, it is important that new control strategies which meet requirements for low toxicity to non-target species, vertebrates and the environment are pursued. This would be greatly facilitated by an improved understanding of the physiology and pharmacology of these nematodes, but to date, these microscopic species of the Phylum Nematoda have attracted little attention in this regard. In this review, the current information available for neurotransmitters and neuromodulator in the plant parasitic nematodes is discussed in the context of the more extensive literature for other species in the phylum, most notably Caenorhabditis elegans and Ascaris suum. Areas of commonality and distinctiveness in terms of neurotransmitter profile and function between these species are highlighted with a view to improving understanding of to what extent, and with what level of confidence, this information may be extrapolated to the plant parasitic nematodes.

Little is known about the fundamental biology of parasitic nematodes (=roundworms) that cause serious diseases, affecting literally billions of animals and humans worldwide. Unlocking the biology of these neglected pathogens using modern technologies will yield crucial and profound knowledge of their molecular biology, and could lead to new treatment and control strategies. Supported by studies in the free-living nematode, Caenorhabditis elegans, some recent investigations have provided improved insights into selected protein phosphatases (PPs) of economically important parasitic nematodes (Strongylida). In the present article, we review this progress and assess the potential of serine/threonine phosphatase (STP) genes and/or their products as targets for new nematocidal drugs. Current information indicates that some small molecules, known to specifically inhibit PPs, might be developed as nematocides. For instance, some cantharidin analogues are known to display exquisite PP-inhibitor activity, which indicates that some of them could be designed and tailored to specifically inhibit selected STPs of nematodes. This information provides prospects for the discovery of an entirely novel class of nematocides, which is of paramount importance, given the serious problems linked to anthelmintic resistance in parasitic nematode populations of livestock, and has the potential to lead to significant biotechnological outcomes.

Under the bioassay-guided method, two diterpenes, 3-O-(2″,3″-dimethylbutanoyl)-13-Ododecanoylingenol (1) and 3-O-(2″,3″-dimethylbutanoyl)-13-O-decanoylingenol (2) isolated from Euphorbia kansui, showed a pronounced antinematodal activity against the nematode Bursaphelenchus xylophilus at the same minimum effective dose (MED) of 5 μg per cotton ball and still displayed antinematodal activity at a dose of 2.5 μg per cotton ball. Compounds 3 − 6 were obtained, and the structure of the new compound 6 was elucidated based on 1D-and 2D-NMR analyses and physicochemical data. Preliminary structure-biological activity relationships of ingenane-type compounds were deduced.

International audience; The increasing prevalence of resistance to anthelmintics among gastrointestinal nematodes and the desire for lower input agriculture have promoted the idea that targeted selective treatment (treating the animals in need of such a treatment and only them) could be a sustainable solution for controlling internal parasites of ruminants. The pros are the slowing of resistance prevalence, lower residues of anthelmintics in meat and milk, and lower cost, the cons are the difficulty and time spent on selecting animals in need of treatment and the possibility of lower production, Using actual experiments and modelling we show that targeted selective treatment can be used to sustainably control gastrointestinal nematode infections in flock.

Pyrantel, a tetrahydropyrimidine nicotinic agonist anthelmintic, has been used in companion animal medicine since the 1970s to control two important nematode groups, the hookworms and the roundworms. Given the zoonotic potential of these parasites, pyrantel has served a dual role in helping to protect the health of both companion animals and the public for more than 30 years. This review describes the history and mechanism of action of this drug, and collates evidence that resistance to pyrantel has developed in at least one canine nematode, the hookworm Ancylostoma caninum. The role of in vitro diagnosis tests in managing anthelmintic resistance in companion animal parasites is discussed, as are management practices that may reduce the rate at which resistance develops.

Aim of the study: The present study was carried out to determine the anthelmintic activity of Caesalpinia crista (L.) (Fabaceae) seed kernel and Chenopodium album (L.) (Chenopodiaceae) whole plant in order to justify their traditional use in veterinary medicine. Materials and methods: In vitro anthelmintic activity of crude aqueous methanolic extract (AME) of both the plants was determined using mature Haemonchus contortus and their eggs in adult motility assay and egg hatch test, respectively. In vivo anthelmintic activity was evaluated in sheep naturally infected with mixed species of gastrointestinal nematodes by administering crude powder (CP) and AME in increasing doses (1.0–3.0 g/kg). Results: Both plants exhibited dose- and time-dependent anthelmintic effects by causing mortality of worms and inhibition of egg hatching. Caesalpinia crista (LC50 = 0.134 mg/mL) was found to be more potent than Chenopodium album (LC50 = 0.449 mg/mL) in egg hatch test. In vivo, maximum reduction in eggs per gram (EPG) of faeces was recorded as 93.9 and 82.2% with Caesalpinia crista and Chenopodium album AME at 3.0 g/kg on day 13 and 5 post-treatment, respectively. Levamisole (7.5 mg/kg), a standard anthelmintic agent, showed 95.1–95.6% reduction in EPG. Conclusions: These data show that both Caesalpinia crista and Chenopodium album possess anthelmintic activity in vitro and in vivo, thus, justifying their use in the traditional medicine system of Pakistan. © 2007 Elsevier Ireland Ltd. All rights reserved.

Due to consumer reluctance to take synthetic drugs for nematode infections and the appearance of resistance to anthelminthic drugs, new drugs from natural products must be developed. Caenorhabditis elegans is one of the freely living nematodes and serves as a good model system for screening anthelminthic drugs. In this study, thirteen flavonoid derivatives were tested for anthelminthic activity and the relationships between their activities and structures were investigated. The structural information combined with the data for the larval growth inhibition of C. elegans provided meaningful structural insights in the search for new anthelminthic drugs.

This paper describes in vitro and in vivo anthelmintic activities of Swertia chirata. In vitro studies revealed that at 25 mg/ml (P ≥ 0.05) the crude aqueous (CAE) and methanolic extracts (CME) of S. chirata whole plant showed an anthelmintic effect on live Haemonchus contortus. Moreover, in the in vivo study, the whole plant of S. chirata administered as crude powder (CP), CAE and CME at the dose of 3 g/kg to sheep naturally infected with mixed species of gastrointestinal nematodes, showed a significant reduction in egg per gram of faeces.

A survey of the prevalence of anthelmintic resistant nematode populations was conducted on 32 sheep farms in the Slovak Republic. In vitro egg hatch test and larval development tests were used for the detection of resistance to benzimidazole anthelmintics and they were compared with in vivo faecal egg count reduction tests. There was agreement in the declaration of resistance between the faecal egg count reduction test and both in vitro tests. The presence of resistant populations was determined on two farms using egg hatch test. In both farms, the LD 50 values were higher than 0.1 μg TBZ/ml, indicating resistance. By using LD 99 values it might be possible to reveal relatively small proportion of resistant larvae in the population. The prevalence of benzimidazole resistance has not change on Slovak sheep farms during last decade.

Human filarial nematodes cause river blindness and lymphatic filariasis, both of which are diseases that produce considerable morbidity. Control of these diseases relies on drug treatments that are ineffective against macrofilariae and are threatened by the development of resistance. New validated drug targets are required to allow development of new classes of antifilarial drugs. To identify and validate potential new drug targets, we propose a collaborative research strategy utilizing bioinformatic filters and assessment of gene function by RNA interference in Caenorhabditis elegans and Brugia malayi.

Among plant pathogens, sedentary endoparasitic nematodes are one of the most damaging pests in global agriculture. These obligate parasites interact with their hosts in a quite unique and intriguing way. They induce the redifferentiation of root cells into specialized feeding cells essential for nematode growth and reproduction; thus, nematodes have evolved the ability to exploit plant genes and hijack host functions for their own requirements. Various approaches to engineer plants with resistance to parasitic nematodes have been pursued, most focusing on the introduction of resistance genes. An alternative strategy to achieve resistance is to exploit the susceptibility of plant disease. Better knowledge of the plant response during the compatible interaction should allow the identification of targets to engineer resistance to parasitic nematodes in crop species.

Levamisole is an anthelmintic agent that exerts its therapeutic effect by acting as a fall agonist of the nicotinic receptor (AChR) of nematode muscle. Its action at the mammalian muscle AChR has not been elucidated to date despite its wide use as an anthelmintic in humans and cattle. By single channel and macroscopic current recordings, we investigated the interaction of levamisole with the mammalian muscle AChR. Levamisole activates mammalian AChRs. However, single channel openings are briefer than those activated by acetylcholine (ACh) and do not appear in clusters at high concentrations. The peak current induced by levamisole is about 3% that activated by ACh. Thus, the anthelmintic acts as a weak agonist of the mammalian AChR. Levamisole also produces open channel blockade of the AChR. The apparent affinity for block (190 μM at -70 mV) is similar to that of the nematode AChR, suggesting that differences in channel activation kinetics govern the different sensitivity of nematode and mammalian muscle to anthelmintics. To identify the structural basis of this different sensitivity, we performed mutagenesis targeting residues in the α subunit that differ between vertebrates and nematodes. The replacement of the conserved αGly-153 with the homologous glutamic acid of nematode AChR significantly increases the efficacy of levamisole to activate channels. Channel activity takes place in clusters having two different kinetic modes. The kinetics of the high open probability mode are almost identical when the agonist is ACh or levamisole. It is concluded that αGly-153 is involved in the low efficacy of levamisole to activate mammalian muscle AChRs. Fil: Rayes, Diego Hernán. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina Fil: de Rosa, Maria Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina Fil: Bartos, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina Fil: Bouzat, Cecilia Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina

The ability of three antinematodal agents to induce paralysis of Caenorhabditis elegans was examined in an aqueous medium with and without the addition of salts. The basic medium was deionized water supplemented, for control purposes, with the phosphate mixture used as a buffering agent in M9 solution. This medium was further supplemented with magnesium sulfate or sodium chloride, or both salts, at the concentrations used in M9. We report that the paralyzing property of ivermectin was enhanced by the presence of salt, while the efficacy of levamisole and chlorpromazine was reduced.

The ability of organisms to evolve resistance threatens the effectiveness of every antibiotic drug. We show that in the nematode Caenorhabditis elegans, simultaneous mutation of three genes, avr-14, avr-15 , and glc-1 , encoding glutamate-gated chloride channel (GluCl) α-type subunits confers high-level resistance to the antiparasitic drug ivermectin. In contrast, mutating any two channel genes confers modest or no resistance. We propose a model in which ivermectin sensitivity in C. elegans is mediated by genes affecting parallel genetic pathways defined by the family of GluCl genes. The sensitivity of these pathways is further modulated by unc-7 , unc-9 , and the Dyf (dye filling defective) genes, which alter the structure of the nervous system. Our results suggest that the evolution of drug resistance can be slowed by targeting antibiotic drugs to several members of a multigene family.

Spectroscopic data define the structures of the flagranones A (2), B (3) and C (4) from the nematode-trapping fungus Duddingtonia flagrans. These antibiotics are structurally related to the farnesylated cyclohexenoxides of the oligosporon group recently isolated from the nematode-trapping fungus Arthrobotrys oligospora, and show similar antimicrobial activity.