Glutathione-S-transferase,a possible drug-metabolizing enzyme,in Haemonchus contortus: Comparative activity of a cambendazole-resistant and a susceptible strain

Kawalek J. C., Rew R. S. and Heavner J. 1984. Glutathione-S-transferase, a possible drug-metabolizing enzyme in Haemonchus contortus: comparative activity of a cambendazole-resistant and a susceptible strain. International Journal for Parasitology14: 173–175. A drug metabolizing enzyme (DME), glutathione-S-transferase, was detected in homogenates of a cambendazole-susceptible and a cambendazole-resistant strain of Haemonchus contortus. The activity was 1.5–1.8 times higher in the resistant strain. DME activation is a possible mechanism for anthelmintic resistance in H. contortus.     

The association between frequency of thiabendazole treatment and the development of resistance in field isolates of Ostertagia spp. of sheep

Martin P. J., Anderson N., Lwin T., Nelson G. and Morgan T. E. 1984. The association between frequency of thiabendazole treatment and the development of resistance in field isolates of Ostertagia spp. of sheep. International Journal for Parasitology14: 177–181. Isolates of Ostertagia spp. were obtained from grazing sheep 3,4 and 5 years after nil, planned (five per year) and regular (3-weekly) treatments with thiabendazole (TBZ). Levels of resistance to TBZ were measured by an in vitro egg hatch assay and a controlled anthelmintic efficiency assay. Isolates from planned treatment groups showed an increase in the level of resistance; the lethal concentrations of TBZ to 50% of eggs (LC₅₀s) were 3, 3 and 6 times the LC₅₀s of isolates from nil treatment groups for years 3, 4 and 5 respectively. The LC₅₀s of isolates from regular treatment groups were 14 times higher than those from nil treatment groups in each year. To assess the potential for an increase in level of resistance, additional egg assays were done 14 days after treatment with 44 mg kg^t?1 of TBZ on sheep infected with the planned group isolates for each year. This treatment raised the LC₅₀S for years 3, 4 and 5 respectively by 3, 2 and 1–5 times the LC₅₀s of the same isolates which had not been exposed to additional TBZ treatment. The controlled anthelmintic efficiency assay using 44 mg kg^t?1 of TBZ produced a significant reduction in the number of adult and immature worms from the nil isolate but failed significantly to reduce the number of worms from the planned and regular isolates. A three component analysis resolved the nonlinear trends of the log dose-probit plots in egg hatch assays for isolates from planned treatment groups into subpopulations of susceptible, hybrid and resistant individuals each with different levels of resistance. The proportions of these subpopulations changed in accordance with the level of resistance observed in each year.     

Development of anthelmintic resistance in nematodes from sheep in Australia subjected to different treatment frequencies

Development of anthclmintic resistance in nematodes from sheep in Australia subjected to different treatment frequencies. International Journal for Parasitology13: 125–132. In a three year (1978, 1979, 1980) field experiment in south-eastern Australia Merino weaner sheep were exposed to one of four worm control programmes. Sheep were treated either 50–51 times, 9–12 times, 3–6 times or 1–3 times annually with anthelmintic. Thiabendazole (TBZ) was used throughout 1978 and for the initial dose in 1980, levamisole (LEV) throughout 1979 and 1980. Sheep were replaced annually each December.During 1978 in vitro egg hatch tests indicated that populations of Haemonchus contenus with resistance factors to TBZ of 10·7, 10·0, 2.5 and 1·8 respectively were selected by these treatment frequencies. No resistance to LEV was detected in 1979 but during 1980 in vivo egg count reduction tests showed that highly resistant populations of Ostertagia spp. were selected in sheep dosed either 49 or 11 times. This was confirmed by an in vitro larval paralysis test. Partial resistance occurred where sheep were dosed 5 times but not where one dose was given. The only population of H. conlortus still present at the end of 1980 in sufficient numbers to allow eggs to be tested in vitro was from the sheep dosed once. This had retained its resistance to TBZ.     

Comparison of efficacy of moxidectin and ivermectin in the treatment of Strongyloides fulleborni infection in rhesus macaques

BACKGROUND: Strongyloides infection may result in clinical disease or confound experimental protocols that utilize non-human primates. There is presently a Strongyloides fulleborni infection rate of approximately 27% in the Tulane National Primate Research Center's breeding colonies despite the routine therapeutic and prophylactic use of ivermectin. METHODS: A study was conducted to determine if moxidectin treatment offers advantages to the intestinal parasite control program. A total of 150 rhesus macaques (Macaca mulatta) that were removed from the breeding colonies due to illness were selected for the study. The animals were randomly assigned to treatment groups with 75 receiving ivermectin and 75 receiving moxidectin. Egg counts were performed on fecal samples collected pre- and post-treatment. RESULTS: Both treatments resulted in decreases in the number of eggs/g in the post-treatment sample as compared with the pre-treatment sample; however, no significant difference was found between treatment groups. CONCLUSIONS: With the data demonstrating a similar efficacy in both ivermectin and moxidectin treated macaques, the benefit of moxidectin treatment relates to biosafety and topical application.     

Effect of particle size on the anthelmintic efficacy of mebendazole against Nippostrongylus brasiliensis in the rat.

The effect of reduction in particle size on the anthelmintic efficacy of mebendazole (methyl 5 (6)- benzoyle 1–2 benzimidazole carbamate) was studied in rats undergoing a primary infection with N. brasiliensis. A single oral treatment with fine ground mebendazole (particle size spectrum—54·95 per cent of particles less than 10·62 μ dia.; 86·06 per cent less than 21·27 μ) removed more than 98 per cent of adult worms from the intestine at a dose rate of 12·5 mg/kg body wt. On the other hand the best result achieved with coarse ground mebendazole (18·47 per cent of particles less than 10·62 μ dia; 42·26 per cent less than 21·27 μ dia) was 58 per cent of adult worms removed at a dose rate of 100 mg/kg body wt. It was also shown that fine ground mebendazole adversely affected migrating third stage larvae of N. brasiliensis.     

Contrasting genetic structures of two parasitic nematodes,determined on the basis of neutral microsatellite markers and selected anthelmintic resistance markers

For the first time, the neutral genetic relatedness of natural populations of Trichostrongylid nematodes was investigated in relation to polymorphism of the β‐tubulin gene, which is selected for anthelminthic treatments. The aim of the study was to assess the contribution of several evolutionary processes: migration and genetic drift by neutral genetic markers and selection by anthelminthic treatments on the presence of resistance alleles at β‐tubulin. We studied two nematode species (Teladorsagia circumcincta and Haemonchus contortus) common in temperate climatic zones; these species are characterized by contrasting life history traits. We studied 10 isolated populations of goat nematode parasites: no infected adult goat had been exchanged after the herds were established. Beta‐tubulin polymorphism was similar in these two species. One and two β‐tubulin alleles from T. circumcincta and H. contortus respectively were shared by several populations. Most of the β‐tubulin alleles were ‘private’ alleles. No recombination between alleles was detected in BZ‐resistant alleles from T. circumcincta and H. contortus. The T. circumcincta populations have not diverged much since their isolation (F_ST <0.08), whereas H. contortus displayed marked local genetic differentiation (F_ST ranging from 0.08 to 0.18). These findings suggest that there are severe bottlenecks in the H. contortus populations, possibly because of their reduced abundance during unfavourable periods and their high reproductive rate, which allows the species to persist even after severe population reduction. Overall, the data reported contradict the hypothesis of the origin of β‐tubulin resistance alleles in these populations from a single mutational event, but two other hypotheses (recurrent mutation generating new alleles in isolated populations and the introduction of existing alleles) emerge as equally likely.     

Structure-Activity Relationship of Anthelmintic Cyclooctadepsipeptides

The relationship between cyclooctadepsipeptides and their anthelmintic efficacy was examined by converting the natural products, PF1022A, PF1022E and PF1022H. Some analogues substituted at the para position of the phenyllactate moiety showed higher or equivalent activity against the parasitic nematode, Ascaridia galli in chicken when compared with the parent compounds. It is suggested that lipophilicity and the polar surface area, in addition to structural requirements of the derivatives, influenced the anthelmintic efficacy in vivo.     

Uptake of thiabendazole and its effects on glucose uptake and carbohydrate levels in the thiabendazole-resistant and susceptible Trichostrongylus colubriformis

Sangster N. C. and Prichard R. K. 1984. Uptake of thiabenzadole and its effects on glucose uptake and carbohydrate levels in thiabendazole-resistant and susceptible Trichostrongylus colubriformis. International Journal for Parasitology14:121–126. Some possible mechanisms of thiabenzadole (TBZ) resistance have been investigated in Trichostrongylus colubriformis. Uptake of TBZ and 3-0-methyl glucose and changes in carbohydrate levels in a TBZ-resistant and a TBZ-susceptible strain of T. colubriformis were measured after in vitro incubation, in the absence or presence of TBZ. The rate of TBZ uptake in adult worms was greater in the resistant strain but the difference was not statistically significant. Final concentrations of TBZ in resistant worms were significantly greater than in the susceptible strain, but there was no difference in TBZ uptake by eggs of either strain. A mechanism of resistance based on decreased drug uptake by resistant parasites is thus unlikely. Resistant worms absorbed 3-0-methyl glucose faster than susceptible parasites, however, this rate was unchanged in either strain when the worms were exposed to TBZ and indicates that the drug does not affect glucose uptake. Levels of glucose, trehalose and glycogen fell during the incubation with the decrease being more pronounced in the resistant strain. When the worms were exposed to TBZ the same changes in carbohydrate levels occurred except for resistant worms maintained anaerobically where levels were higher, showing that TBZ does not affect the rate of carbohydrate utilization.     

Trichostrongylus colubriformis: effect of anthelmintics on ingestion and oviposition

The effects of the anthelmintics ivermectin, levaminsole, oxfendazole, piperazine citrate, pyrantel pamoate, tetramisole, and thiabendazole on ingestion and oviposition by Trichostrongylus colubriformis were determined. Six of the compounds reduced in vitro feeding at the tested doses while all drugs reduced in vivo feeding after treatment of the host. Additionally, in vitro or in vivo exposure to most anthelmintics decreased oviposition during subsequent in vitro assay. Invermectin had the most pronounced effect on in vivo and in vitro feeding and egg release. The neuromuscular activities of pharyngeal pumping and egg ejection may be suitable systems for rapid determinations of anthelmintic effects.     

Insecticidal and nematicidal properties of microbial metabolites

Summary Metabolites from 942 microbial isolates were screened for insecticidal and nematicidal properties. The isolates included 302 streptomycetes, 502 novel actinomycetes including representatives of 18 genera, 28 unidentified aerobic actinomycetes, 70 fungi and 40 bacteria other than actinomycetes. When diluted 10-fold, the metabolites from 55 isolates caused nearly 100% mortality in mosquito larvae (Aedes aegypti) within 24 h. These isolates included 27 isolates ofStreptomyces, four ofActinoplanes, three isolates each ofActinomadura andStreptoverticillium, two isolates each ofMicromonospora, Bacillus andPaecilomyces, and one isolate each ofMicropolyspora, Nocardiopsis, Streptosporangium, Oerskovia, Thermomonospora, Chainia, Pseudomonas, Fusarium, Monilia andSyncephalestrum. Two fungal isolates could not be identified to the generie level. Extracts from the culture broth of 18 isolates caused 100% mortality in mosquito larvae within 15 min to 24 h at a concentration of 1000 ppm. The LC₅₀ of partially purified products from two isolates was 1–2.5 ppm and that of the semipurified preparations from four other isolates was 50 ppm. Valinomycin was identified as an active component in the culture broth from one isolate. The culture broth from 15 isolates of aerobic actinomycetes and 4 of fungi were toxic toPanagrellus redivivus (Nematoda); these included 12 isolates with selective nematicidal properties.Michigan Agriculture Experiment Station, Journal Article No. 12321.     

The effect of selection with levamisole on benzimidazole resistance in Ostertagia spp. of sheep

A field population of Ostertagia spp. (predominantly O. circumcincta) in sheep, found to be resistant to oxfendazole, was exposed to selection with1 levamisole in the laboratory and the field. Progeny of the survivors of a single dose of levamisole in penned sheep showed a significantly lower level of resistance to oxfendazole in an anthelmintic assay, and a similarly lower level of resistance to the ovicidal activity of thiabendazole in vitro compared with the progeny of the survivors of a single dose of oxfendazole, although they remained more resistant than a known susceptible strain of 0. circumcincta. The progeny of worms present in grazing sheep after three doses of levamisole at 4-weekly intervals, and subjected to the same tests, behaved in a similar fashion. They also showed a substantial fall in benzimidazole resistance compared with the progeny of worms not exposed to any anthelmintic for 6 months. The results suggested that levamisole selected positively against benzimidazole resistance. In these circumstances some form of alternation in the use of different anthelmintics could delay the development of a high level of resistance and maintain the practical usefulness of existing anthelmintics.     

Synthetic Aurones: New Features for Schistosoma mansoni Therapy

In this work, two synthetic aurones revealed moderate schistosomicidal potential in in vitro and in vivo assays. Aurones ( 1 ) and ( 2 ) promoted changes in tegument integrity and motor activity, leading to death of adult Schistosoma mansoni worms in in vitro assays. When administered orally (two doses of 50 mg/kg) in experimentally infected animals, synthetic aurones ( 1 ) and ( 2 ) promoted reductions of 56.20 % and 57.61 % of the parasite load and stimulated the displacement towards the liver of the remaining adult worms. The oogram analysis revealed that the treatment with both aurones interferes with the egg development kinetics in the intestinal tissue. Seeking an action target for compounds ( 1 ) and ( 2 ), the connection with NTPDases enzymes, recognized as important therapeutic targets for S. mansoni, was evaluated. Molecular docking studies have shown promising results. The dataset reveals the anthelmintic character of these compounds, which can be used in the development of new therapies for schistosomiasis.     

Development and validation of an in vitro Trichostrongylus colubriformis motility assay

Development and validation of an in vitro Trichostrongylus colubriformis motility assay. International Journal for Parasitology 17: 1441–1444. An in vitro Trichostrongylus colubriformis motility assay involving the use of a micromotility meter has been developed and validated. Four commercially available ruminant anthelmintics (albendazole, ivermectin, levamisole hydrochloride, and coumaphos) and an investigational hydrazone compound (p-toluoyl chloride phenylhydrazone) were evaluated in this assay at four concentrations each. At 100 μg ml^-1, all five treatments significantly (P 0.05) reduced the motility of ensheathed L-3 T. colubriformis larvae, thereby indicating anthelmintic activity. At this concentration, coumaphos was significantly less active than any of the other four treatments. At 10 μg ml^-1 albendazole, ivermectin, levamisole hydrochloride and the hydrazone compound were active, but coumaphos was not. At 1 μg ml^-1 albendazole, ivermectin and levamisole hydrochloride remained significantly active, but neither coumaphos nor the hydrazone compound showed significant activity. At all three of the higher concentrations (1,10 and 100 μg ml^-1), levamisole hydrochloride indicated greater activity than any of the other treatments. This difference was statistically significant at the 1 and 10 μg ml^-1 concentrations. None of the five treatments showed significant activity at the lowest concentration (0.1 μg ml^-1). The in vitro T. colubriformis motility assay proved to be sensitive, accurate, rapid, and repeatable. This assay system should be another valuable addition to the tests used to identify potential anthelmintics, monitor helminth resistance to drugs, and define the kinetics and mode of action of drugs.     

A laboratory assessment of the potential molluscicidal activity of some Nigerian plant species used as anthelmintics

Laboratory studies were conducted to determine the potential molluscicidal activity of five Nigerian plant species used in alternative medicine practice as anthelmintics. Laboratory-bred adult Biomphalaria pfeifferi, the snail host of Schistosoma mansoni in Nigeria, and their viable 0- to 24-hour-old egg masses were separately exposed for 24 hours, in replicated bioassays, to 7.81–4 000 mg l^?1 concentrations of crude ethanolic extracts of the leaves and roots of Annona senegalensis, Anogeissus leiocarpus, Crotalaria retusa, Dalbergia sissoo and Vernonia amygdalina. Fruits and stem bark of D. sissoo were also investigated. The 24-hour LC50 and LC90 values of extracts for target organisms were calculated using probit analysis. Susceptibility of target organisms to extracts varied with the plant species and morphological part. Egg masses and adult snails were most susceptible to D. sissoo fruit extract (LC90 values of 89.29 and 74.33 mg l^?1, respectively) and least susceptible to V. amygdalina leaf extract (LC90 values of 11 864.09 and 6 241.61 mg l^?1, respectively). Concentration-dependent behavioural responses and embryo deaths at the gastrula/exogastrula and/or prehatch snail stages of development were observed in exposed snails and egg masses, respectively. Tested extracts of A. senegalensis, A. leiocarpus, C. retusa and V. amygdalina did not show appreciable toxicities to B. pfeifferi egg masses or adults. For demonstrating significant molluscicidal activity (LC90 < 100 mg l^?1) and toxicities to egg masses, we recommend further studies on the ethanolic extracts of D. sissoo fruits and roots.