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Evaluation of anthelmintic properties of extracts from some plants used as livestock dewormers by pastoralist and smallholder farmers in Kenya against Heligmosomoides polygyrus infections in mice.

Parasitic nematodes are among the most common and economically important infectious diseases of grazing livestock, especially in small ruminants in the tropics and subtropics in Kenya the control of gastrointestinal nematode infections in sheep and goats is usually made with synthetic anthelmintics but substantial levels of anthelmintic resistance have been recorded. A number of medicinal plants, that may provide possible alternatives, and are used by pastoralists and smallholder farmers in Kenya as deworming agents for their livestock and equines, namely Aframomum sanguineum, Dodonea angustifolia, Hildebrandtia sepalosa, Myrsine africana, Rapanea melanophloeos from Kenya, and Azadirachta indica from Kenya and Malaysia, together with the chemicals embelin and santonin that occur in some of these plants, were evaluated against Heligmosomoides polygyrus in mice. Commercial anthelmintics, namely ivermectin, pyrantel and piperazine, were also investigated, both to validate the mouse model system and to assess efficacy of these drugs against H. polygyrus. Pyrantel and ivermectin were highly effective in reducing the numbers of H. polygyrus worms as well as eggs in faeces of the mice, but piperazine had a lower activity. Application of santonin and M. africana significantly reduced the number of total worm counts (TWC) but not faecal egg counts (FEC). The use of embelin, R. melanophloeos and A. indica reduced FEC but not TWC. In all cases, however, reductions were well below the a priori level of 70% required for biological significance. A. sanguineum, D. angustifolia and H. sepalosa had no effect on either TWC or FEC. In conclusion, none of the plant preparations had any biologically significant anthelmintic effect in this monogastric host-parasite model system.

Animals↗

Larval development assay for detection of anthelmintic resistance in cyathostomins of Swedish horses.

The aim of this study was to investigate the suitability of a larval development assay (LDA) for the determination of anthelmintic resistance in cyathostomin nematode populations of the horse. In addition, comparison of results between geographic regions, types of horse establishment, and the use of anthelmintics in Sweden, was established. Seventy horse herds from different parts of Sweden were sampled, and strongyle eggs from the faeces of 54 of those were investigated by an LDA (DrenchRite). The following anthelmintics were tested: thiabendazole (TBZ), levamisole (LEV), ivermectin monosaccharide (IVM-MS), ivermectin aglycone (IVM-AG) and pyrantel (PYR). The LC50 values for TBZ and LEV were generally lower than those previously reported in other LDA studies on horse nematodes. This could be related to the infrequent use of these compounds for the past 20 years in Sweden. In this study, there was a great variation within and between assay plates that could not be explained. Still the LC50 values differed significantly between the regions for all anthelmintics, except for pyrantel. The highest LC50s were observed in parasite populations from the south of Sweden. There were no significant differences between riding schools and studs. Limitations of this technique exist, namely the lack of established cut-off values for susceptible and resistant populations and interpretation problems related to multi-species infections. Although there are advantages with LDA such as the possibility of testing several compounds simultaneously without interference with the deworming programmes on the farms, we conclude that LDA currently is not a reliable alternative to the faecal egg count reduction test (FECRT).

Animals↗

Prescription-only anthelmintics--a questionnaire survey of strategies for surveillance and control of equine strongyles in Denmark.

In 1999, legislation in Denmark made anthelmintic drugs available only by prescription, and prohibited their use for routine, prophylactic treatment. A questionnaire survey was conducted in 2004 to determine current strategies for surveillance and control of equine strongyles used in Danish equine veterinary practices. Eighty-seven of 170 (51.2%) registered equine veterinary practices responded. Ninety seven percent of the respondents used faecal egg counts for diagnosis and surveillance, and 41% used larval cultures. Logistic regression revealed that the use of larval cultures was positively correlated with numbers of employees (P = 0.013) and the proportion of equine caseload in the practice (P < 0.000). Performing faecal egg counts and treating horses was most frequent during spring, summer and early autumn. Veterinary practices made treatment decisions based on cut-off values ranging from 20 to 500 eggs per gram (EPG) faeces. Ages of horses, clinical suspicions of parasitic disease, or requests by the owners were the most important factors in the strategies for faecal sampling. Less commonly, sampling strategies included all horses on the premises and random sampling. Ninety five percent of the respondents reported treating certain groups of horses without prior faecal analysis, including horses with clinical signs of parasitic disease (77%), foals (84%), horses less than 3 years of age (52%), and pregnant mares (51%). The respondents regarded the cyathostomin group as the most prevalent cause of parasitic disease and ill-thrift, followed by large strongyles and Parascaris equorum. Sixty seven percent rotated regularly between drugs, while 11% performed routine screening for anthelmintic resistance. Results of this survey suggest that limiting equine anthelmintics to prescription-only availability has increased the level of strongyle surveillance. Veterinary practitioners play a central role in equine parasite management with indications of a lowered intensity of treatment. However, screening for anthelmintic resistance remains uncommon.

Age Factors↗

The detection of anthelmintic resistance in nematodes of veterinary importance.

Before revised World Association for the Advancement of Veterinary Parasitology (WAAVP) guidelines on the detection of anthelmintic resistance can be produced, validation of modified and new methods is required in laboratories in different parts of the world. There is a great need for improved methods of detection of anthelmintic resistance particularly for the detection of macrocyclic lactone resistance and for the detection of resistant nematodes in cattle. Therefore, revised and new methods are provided here for the detection of anthelmintic resistance in nematodes of ruminants, horses and pigs as a basis for discussion and with the purpose that they are evaluated internationally to establish whether they could in the future be recommended by the WAAVP. The interpretation of the faecal egg count reduction test has been modified and suggestions given on its use with persistent anthelmintics and continuous release devices. An egg hatch test for benzimidazole (BZ) resistance is described. A microagar larval development test for the detection of benzimidazole and levamisole resistance provides third stage larvae for the identification of resistant worms. The sensitivity of these two tests can be increased by using discriminating doses rather than LD(50) values. Details are given of a PCR based test for the analysis of benzimidazole resistance in strongyles of sheep and goats, horses and cattle. Although promising for ruminant trichostrongyles, quantitative determination of gene frequency using real time PCR requires further development before PCR tests will be used in the field. Apart from faecal egg count reduction tests there are currently no satisfactory tests for macrocylic lactone resistance despite the great importance of this subject. Except for treatment and slaughter trials there are no validated tests for fasciolicide resistance or for the detection of resistance in cestodes.

Animals↗

The effect of mixed or sequential grazing of cattle and sheep on the faecal egg counts and growth rates of weaned lambs when treated with anthelmintics.

An experiment was conducted over two successive years (2002 and 2003) to investigate the effects of grazing improved permanent pasture (mainly perennial ryegrass/white clover) by cattle or sheep, either sequentially or mixed, on the faecal egg counts and growth rates of weaned lambs when treated with anthelmintics. The grazing season was divided into two parts, May-July then July-October, relating to the pre- and post-weaning of the lambs. Four grazing regimes, replicated three times, were compared: (1) sheep only from May to October (SS); (2) cattle May to July followed by lambs until October (C/S); (3) cattle and sheep May to July followed by lambs until October (C+S/S); and (4) cattle and sheep May to July followed by cattle and lambs until October (C+S/C+S). Sward height was maintained at 6 cm using a "put and take" stocking system. At weaning, lambs were weighed and treated with an anthelmintic (0.08% ivermectin drench, Oramec) before being allocated to plots. They were then weighed and drenched every 28 days until the end of the experiment (Day 84). Faecal egg counts (FEC) were measured in all lambs immediately prior to each anthelmintic treatment. In 2002 and 2003, there were differences between the groups in FEC, with the SS lambs having the highest values and C/S lambs the lowest (P<0.01). There were also differences in the rate of liveweight gain of the lambs in each of the study years: for this parameter SS lambs had the lowest growth rate but the fastest growth was in C+S/C+S lambs not C/S lambs (P<0.01), indicating that these differences were due to factors other than parasite infection. Overall, sequential grazing of pastures with cattle then sheep reduced the faecal egg counts in lambs regularly treated with anthelmintics when compared with lambs grazing in mixed systems with cattle and sheep or with sheep only systems, however, the highest growth rates were observed in lambs in the mixed cattle/sheep grazing system throughout.

Animal Husbandry↗

The biochemical basis of anthelmintic action and resistance.

The most commonly used modern anthelmintics include the benzimidazoles, the nicotinic agonists. praziquantel, triclabendazole and the macrocyclic lactones. These drugs interfere with target sites that are either unique to the parasite or differ in their structural features from those of the homologous counterpart present in the vertebrate host. The benzimidazoles exert their effect by binding selectively and with high affinity to the beta-subunit of helminth microtubule protein. The target site of the nicotinic agonists (e.g. levamisole, tetrahydropyrimidines) is a pharmacologically distinct nicotinic acetylcholine receptor channel in nematodes. The macrocyclic lactones (e.g. ivermectin, moxidectin) act as agonists of a family of invertebrate-specific inhibitory chloride channels that are activated by glutamic acid. The primary mode of action of other important anthelmintics (e.g. praziquantel, triclabendazole) is unknown. Anthelmintic resistance is wide-spread and a serious threat to effective control of helminth infections, especially in the veterinary area. The biochemical and genetic mechanisms underlying anthelmintic resistance are not well understood, but appear to be complex and vary among different helminth species and even isolates. The major mechanisms helminths use to acquire drug resistance appear to be through receptor loss or decrease of the target site affinity for the drug. Knowledge on the mechanisms of drug action and resistance may be exploitable for the development of new drugs and may provide information on ways to overcome parasite resistance, respectively.

Animals↗

Mechanism-based screening: discovery of the next generation of anthelmintics depends upon more basic research.

The therapeutic arsenal for the control of helminth infections contains only a few chemical classes. The development and spread of resistance has eroded the utility of most currently available anthelmintics, at least for some indications, and is a constant threat to further reduce the options for treatment. Discovery and development of novel anthelmintic templates is strategically necessary to preserve the economic and health advantages now gained through chemotherapy. As the costs of development escalate, the question of how best to discover new drugs becomes paramount. Although random screening in infected animals led to the discovery of all currently available anthelmintics, cost constraints and a perception of diminishing returns require new approaches. Taking a cue from drug discovery programmes for human illnesses, we suggest that mechanism-based screening will provide the next generation of anthelmintic molecules. Critical to success in this venture will be the exploitation of the Caenorhabditis elegans genome through bioinformatics and genetic technologies. The greatest obstacle to success in this endeavour is the paucity of information available about the molecular physiology of helminths, making the choice of a discovery target a risky proposition.

Animals↗

The effect of different anthelmintic treatment regimens combined with iron supplementation on the nutritional status of schoolchildren in KwaZulu-Natal, South Africa: a randomized controlled trial.

A randomized controlled trial in KwaZulu-Natal (South Africa) of 428 primary-school pupils (stratified into 6 groups by age, sex and intervention) measured the effect of different anthelmintic treatments and iron supplementation regimens provided twice at 6-monthly intervals for 1 year (1996/97). Half the pupils received iron supplementation (ferrous fumarate 200 mg weekly for 10 weeks). Pupils received 2 anthelmintic regimens, either (i) albendazole 400 mg plus praziquantel 40 mg/kg or (ii) albendazole 400 mg on 3 consecutive days plus praziquantel 40 mg/kg or (iii) placebo. Baseline prevalences of Ascaris 55.9%, Trichuris 83.6%, hookworm spp. 59.4%, were reduced after 12 months for single-dose albendazole treatment to Ascaris 17.4% (P < 0.005), Trichuris 61.5% (NS), hookworm spp. 0% (P < 0.005), and for triple-dose albendazole treatment to Ascaris 14.8% (P < 0.005), Trichuris 25.0% (P < 0.01), hookworm 0% (P < 0.005). Schistosoma haematobium 43.4% was reduced among treated groups to 8.3% (P < 0.005). There were no significant changes in the anthropometry of the different treatment groups at either 6 or 12 months post treatment. Twelve months after treatment there was a significant increase in haemoglobin levels (P = 0.02) among pupils receiving triple-dose albendazole, praziquantel and ferrous fumarate; pupils receiving no anthelmintic treatment showed a significant decrease as did pupils who received triple-dose albendazole and praziquantel but no iron. Regular 6-monthly anthelmintic treatment significantly reduced the prevalence of Ascaris, hookworm spp. and S. haematobium infections (P < 0.05). Triple-dose treatment for Trichuris was significantly more effective than a single dose of albendazole 400 mg (P = 0.002). In areas with schistosomiasis, hookworm infection and high prevalence of Trichuris infection, combination treatment with praziquantel, triple-dose albendazole, plus iron supplementation, is likely to improve pupils' health and haemoglobin levels.

Adolescent↗

Pharmacology of anthelmintic resistance.

Anthelmintic resistance has grown from a curiosity to an important economic problem in several animal industries and is now set to threaten the control of human parasites. The pharmacology of anthelmintics and anthelmintic resistance has been studied most extensively in the nematode parasites of sheep. Here, Nick Sangster and Jenny Gill discuss this veterinary experience, summarizing the progress made in understanding anthelmintic resistance and highlighting the tools available for research.

Animals↗

Resistance to therapeutic treatment with macrocyclic lactone anthelmintics in Ostertagia circumcincta.

Eighty-eight lambs were allocated to one of four groups which were then dosed with 10,000 infective-stage larvae (L3) of one of four populations of Ostertagia circumcincta; the first (S) was an isolate known to be anthelmintic-susceptible; the second (OR) was a multiple anthelmintic-resistant isolate which had been recovered from the field following therapeutic failure of both ivermectin and moxidectin and subsequently maintained in the laboratory without further anthelmintic selection. The third (R) was derived from OR but had been passaged for five generations in the laboratory with each generation being screened with all three broad-spectrum drench families; the fourth (R x S) was an F1 cross between the S and R isolates. On patency, each of the four infection groups was sub-divided into five treatment groups, one of which received no anthelmintic while the others were administered either oral ivermectin (IVM-oral), controlled-release capsules containing ivermectin (IVM-CRCs), oral moxidectin (MOX-oral) or injectable MOX (MOX-inj). Neither formulation of IVM reduced FEC in the R, R x S and OR infected lambs compared to their untreated controls, but significant reductions were observed in all cases following MOX-oral or MOX-inj treatment. Similarly, neither IVM formulation significantly reduced the numbers of R or R x S worms compared to their untreated controls, although the numbers of OR worms were reduced in both cases (P<0.05). Direct comparisons of efficacy across the isolates, however, indicated that neither formulation was any more effective against R x S or OR worms than against the more highly selected R worms. In contrast, both MOX formulations significantly reduced worm numbers of all the resistant isolates compared to their respective untreated controls; furthermore, worm burdens of R x S were reduced significantly more than burdens of R (P<0.05). Reductions in OR burdens, which were intermediate between the two, did not differ significantly from either. The results are consistent with published work on Haemonchus contortus, which suggests that macrocyclic lactone (ML) resistance is expressed as a dominant trait under treatment with IVM. However, these data differ from the H. contortus studies in suggesting that ML resistance in O. circumcincta may effectively be rendered incompletely dominant or recessive by treatment with MOX.

Administration, Oral↗

Targeted application of anthelmintics to control trichostrongylosis in dairy goats: result from a 2-year survey in farms.

The targeted application of anthelmintic treatments represents one of the current available solutions to slow down the development of anthelmintic resistance within worm populations. Within an experimental flock of dairy goats, control of gastrointestinal parasitism by such selective treatments, targeting the most receptive animals within a flock, was previously found to be effective and to have no detrimental consequences on milk production. The objectives of the current study were to verify the validity of this method in farm conditions. Eleven dairy goat farms from three main areas of production in France were surveyed for 2 years. In six farms, the survey was prolonged for a third year. During year 1, systematic treatments were applied during the grazing season whereas in year 2 and or year 3, treatments were given exclusively to the goats in first lactation and to the multiparous ones with the highest potential of milk production. The level of nematode infection was measured four times per year through individual coproscopical examinations and the mean annual production of milk was recorded. No significant changes in egg excretion nor in milk production were noticed in any farm during years 2 or 3 after switching from the systematic to the selective mode of treatments. These results confirmed that targeted application of anthelmintics might represent a way to combine the control of trichostrongyles and the prevention of anthelmintic resistance.

Animals↗

In vitro screening of two Nigerian medicinal plants (Vernonia amygdalina and Annona senegalensis) for anthelmintic activity.

Vernonia amygdalina and Annona senegalensis, two plants used by local/smallholder livestock farmers in Nigeria as anthelmintic were screened for in vitro anthelmintic activity using Haemonchus contortus eggs. The extract of V. amygdalina did not show any significant activity at concentrations up to 11.2mg/ml. The extract of A. senegalensis showed significant (P<0.001) reduction in egg hatch at a concentration of 7.1mg/ml. The in vitro fecal culture of eggs with A. senegalensis showed significant (P<0.001) reductions in larval recovery with increasing concentrations from 1 to 10% ((w/w) in fecal culture) when whole ground plant material was used. Although traditional veterinary healers and local livestock farmers claim both plants are effective as anthelmintics, our result indicate that only A. senegalensis showed promising anthelmintic activity especially with use of ground whole plant materials as used by some farmers.

Animals↗

Frontiers in anthelmintic pharmacology.

Research in anthelmintic pharmacology faces a grim future. The parent field of veterinary parasitology has seemingly been devalued by governments, universities and the animal industry in general. Primarily due to the success of the macrocyclic lactone anthelmintics in cattle, problems caused by helminth infections are widely perceived to be unimportant. The market for anthelmintics in other host species that are plagued by resistance, such as sheep and horses, is thought to be too small to sustain a discovery program in the animal health pharmaceutical industry. These attitudes are both alarming and foolish. The recent history of resistance to antibiotics provides more than adequate warning that complacency about the continued efficacy of any class of drugs for the chemotherapy of an infectious disease is folly. Parasitology remains a dominant feature of veterinary medicine and of the animal health industry. Investment into research on the basic and clinical pharmacology of anthelmintics is essential to ensure chemotherapeutic control of these organisms into the 21st century. In this article, we propose a set of questions that should receive priority for research funding in order to bring this field into the modern era. While the specific questions are open for revision, we believe that organized support of a prioritized list of research objectives could stimulate a renaissance in research in veterinary helminthology. To accept the status quo is to surrender.

ATP Binding Cassette Transporter, Subfamily B↗

Parasite diversity and anthelmintic resistance in two herds of horses.

Diversity of parasite populations was compared between two herds of horses, one a regularly treated herd the other a feral herd which has bad no anthelmintic treatment for at least 25 years. Eggs obtained from fecal samples of both herds were tested for anthelmintic resistance by use of an in-vitro larval hatch/development assay (LDA), DrenchRite. A fecal egg reduction test was also performed with the domesticated herd using fenbendazole, pyrantel pamoate and ivermectin. Cyathostomes were the predominant group of worms present in both herds. Trichostrongylus axei was seen in both herds, but Strongylus equinus, Strongylus vulgaris, Gyalocephalus capitatus, Poteriostomum spp. and Strongyloides westeri were only found in the feral horses. Larvae of Strongylus edentatus were found in a single domesticated horse. Fecal egg reduction tests with the domesticated herd showed a 32% egg count reduction for fenbendazole, a 93% reduction with pyrantel, and a 99% reduction with ivermectin. From the LDA, anthelmintic resistance was evaluated by determining the resistance ratio of the domesticated herd compared with the feral herd. For benzimidazoles in the domesticated herd, 45% of the cyathostome population was 9.4 times more tolerant than the feral herd's parasite population. The parasite population in the domesticated herd was 1.5 times more tolerant to Levamisole, and 1.7 times more tolerant to the benzimidazole/levamisole combination than the parasite population within the feral herd. 9% of the parasite population in the domesticated herd was 90 times more tolerant to avermectins than the feral herd's parasite population, even though a subpopulation of worms in the feral herd were tolerant to low concentrations of avermectins despite never being previously exposed to this class of anthelmintic.

Animals↗

Acetylcholinesterase in infective-stage larvae of Haemonchus contortus, Ostertagia circumcincta and Trichostrongylus colubriformis resistant and susceptible to benzimidazole anthelmintics.

The amounts of acetylcholinesterase (AChE) in homogenates of infective-stage larvae of strains, from different countries, of Haemonchus contortus, Ostertagia circumcincta and Trichostrongylus colubriformis resistant or susceptible to benzimidazole (BZ) anthelmintics, were compared using a rapid colorimetric assay. The levels of AChE were much greater in those strains which were BZ-resistant than in BZ-susceptible populations. The larvae of susceptible and resistant H. contortus contained less AChE than did the larvae of the other two species. One-way analysis of variance showed that the observed differences between susceptible and resistant strains were significant in each of the species studied. It is suggested that the extra enzyme may act as an anthelmintic-detoxifying enzyme, may inactivate the active site of the anthelmintic or may prevent binding of the anthelmintic to its receptor.

Acetylcholinesterase↗

Potential remedy against Echinococcus multilocularis in wild red foxes using baits with anthelmintic distributed around fox breeding dens in Hokkaido, Japan.

The effect of bait-delivered anthelmintic to reduce the prevalence of Echinococcus multilocularis in wild red foxes was evaluated in Koshimizu, in the eastern part of Hokkaido, Japan. The study area (200 km2) was divided into baited and non-baited sections. The anthelmintic baits were distributed around fox den sites in the baited section every month for 13 months. After 1 year of the anthelmintic bait distribution, the prevalence of E. multilocularis in foxes, evaluated either by the parasite egg examination (from 27.1 to 5.6%) or coproantigen ELISA (from 59.6 to 29.7%), decreased in the baited section contrasting to that in the non-baited section (parasite egg: from 18.8 to 24.2%; ELISA: from 41.9 to 45.8%). The prevalence of E. multilocularis in grey red-backed vole Clethrionomys rufocanus, caught around fox dens, born after bait distribution also decreased and was significantly lower than that in non-baited section. However, within the study periods, the coproantigen-positive rate in fox faeces sporadically increased, while egg-positive rate constantly decreased. Since coproantigen ELISA can detect pre-patent infection, this observation indicates that reinfection pressure in the baited section was still high even after the 13 months of anthelmintic bait distribution. Therefore, the bait distribution longer than our study period is required for the efficient control of E. multilocularis in wild red fox population.

Animals↗

Efficacy of two cyclooctadepsipeptides, PF1022A and emodepside, against anthelmintic-resistant nematodes in sheep and cattle.

Resistance against the major currently available anthelmintics has reached a critical level in many small ruminant herds world-wide, and is increasingly becoming a problem in horses and cattle. Therefore, new products with different modes of action are urgently needed. Recently, such a new class of compounds, the anthelmintically active cyclooctadepsipeptides, was described. Here, the effects of cyclooctadepsipeptides on benzimidazole-, levamisole- and ivermectin-resistant populations of Haemonchus contortus in sheep as well as an ivermectin-resistant Cooperia oncophora population in cattle were studied. Experimentally infected sheep and cattle were used. Animals were treated orally, subcutaneously, or intravenously with cyclooctadepsipeptides. The anthelmintic effects were assessed by means of fecal egg count reductions and/or worm count reductions. Both, PF1022A and emodepside were found to be fully effective against these parasite populations. These findings confirm that this new class of compounds acts by a different mode of action compared to the above-mentioned anthelmintics.

Animals↗

Syntheses and anthelmintic activity of alkyl 5(6)-(substituted-carbamoyl)- and 5(6)-(disubstituted-carbamoyl)benzimidazole-2-carbamates and related compounds.

A number of alkyl 5(6)-(substituted-carbamoyl)- and 5(6)-(disubstituted-carbamoyl)benzimidazole-2-carbamates and related compounds have been synthesized, and their anthelmintic activity against various intestinal helminths of experimental animals have been evaluated. A large percentage of the compounds synthesized showed noteworthy activity against Ancylostoma ceylanicum and at higher doses against Hymenolepsis nana infections. Compared to the alkyl 5(6)-(substituted-carbamoyl)benzimidazole-2-carbamates, the disubstituted carbamoyl analogues were found to exhibit better anthelmintic activity. The most active compound of the series, namely, methyl 5(6)-[(N-2-pyridylpiperazino)carbamoyl]benzimidazole-2-carbamate (90), has been screened against intestinal helminths in higher animals and as a micro- and macrofilaricidal agent. Compound 90 has been identified as a broad-spectrum anthelmintic agent. Compound 90 has been identified as a broad-spectrum anthelmintic in view of its efficacy against A. ceylanicum (hamsters and dogs), H. nana (rats), Nippostrongylus brasiliensis (rats), Syphacia obvelata (mice), A. tubaeformis (cat), Toxocara spp. (cat), and Litomosoides carinii (cotton rat).

Ancylostomiasis↗