Development of a novel in vitro equine anthelmintic assay

An in vitro assay involving the use of a horse strongyle (Strongylus edentatus) and the micromotility meter has been developed to test for equine anthelmintic activity. Three commercially available equine anthelmintics (dichlorvos, ivermectin, and pyrantel pamoate) and an investigational drug (p-toluoyl chloride phenylhydrazone) were evaluated in this assay at four concentrations. After a 24-h incubation, greater than or equal to 10 micrograms/ml of all four drug treatments significantly (P less than or equal to 0.05) reduced the motility of ensheathed L-3 S. edentatus larvae, thereby indicating anthelmintic activity. Pyrantel pamoate also reduced motility at 1 microgram/ml, while the hydrazone significantly increased movement at this level. At 0.1 microgram/ml, none of the treatments significantly reduced motility; one treatment (dichlorvos) significantly increased larval motility. Incubation for 48 h resulted in significant activity (reduction in motility) at greater than or equal to 1 microgram/ml with two drugs (ivermectin, pyrantel pamoate); dichlorvos and the hydrazone reduced motility at greater than or equal to 10 micrograms/ml. None of the treatments significantly reduced motility at the lowest concentration (0.1 microgram/ml); however, at 48 h, two treatments (dichlorvos, hydrazone) significantly increased motility at the lowest concentration (0.1 microgram/ml). The in vitro S. edentatus motility assay proved to be sensitive, accurate and rapid. This assay system should be a valuable addition to tests used to identify potential equine anthelmintics, monitor helminth resistance to drugs, and perhaps define the kinetics and mode of action for drugs.     

Efficacy of ivermectin as an anthelmintic in leopard frogs.

Ivermectin administered cutaneously at dosages of 2 mg/kg of body weight eliminated nematode infections in leopard frogs. Three clinical trials were conducted. In the first trial, 5 groups of 11 frogs were given ivermectin IM at dosages of 0, 0.2, 0.4, 2, or 20 mg/kg. All frogs given ivermectin IM at dosages of 2.0 mg/kg or greater died. In trial 2, 44 frogs, allotted to 5 groups, were given ivermectin cutaneously at 0, 0.2, 2, or 20 mg/kg. Cutaneously administered ivermectin was not toxic at dosages up to 20 mg/kg. In trial 3, nematode infections were eliminated in all 10 frogs treated cutaneously with ivermectin at 2.0 mg/kg.     

Getah virus as an equine pathogen.

Getah virus is a member of the genus Alphavirus in the family Togaviridae and has been frequently isolated from mosquitoes. Seroepizootiologic studies indicate that the virus is mosquito-borne and widespread, ranging from Eurasia to southeast and far eastern Asia, the Pacific islands, and Australasia. The natural host animal of the virus was not known until the first recognized occurrence of Getah virus infection among racehorses in two training centers in Japan in 1978. Outbreaks of clinical disease due to Getah virus infection occur infrequently, and only one outbreak has been reported outside Japan; this was in India in 1990. Clinical signs of the disease are mild and nonlife-threatening and are characterized by pyrexia, edema of the hind limbs, swelling of the submandibular lymph nodes, and urticarial rash, as reported in the 1978 epizootic. The morbidity was 37.9% (722 of 1903 horses) in one training center, with 96% of 722 affected horses making a full clinical recovery within a week without any significant sequelae. Antibodies against Getah virus were detected in 61.2% (172 of 281) and 55.8% (254 of 455) of horses at two training centers, respectively. Virus isolation can be attempted in VERO, RK-13, BHK-21, and many other cell lines as well as in suckling mouse brain. Blood plasma collected from suspect cases of infection at the onset of pyrexia is the specimen of choice. A diagnosis of Getah virus infection can also be confirmed serologically based on testing acute and convalescent phase sera by using SN, CF, HI, and ELISA tests. An inactivated vaccine is available for the prevention and control of Getah virus infection in horses in Japan.     

Evaluation of fenbendazole as an extended anthelmintic treatment regimen for swine

Fenbendazole was given in the feed to swine at a cumulative dosage of 9 mg/kg of body weight over a period of 3, 6, and 12 days to compare efficacy. Four treatment groups of ten 2- to 3-month-old pigs each, with a mean of 15 kg of body weight per group, received 3 mg of fenbendazole/kg/day for 3 days, 1.5 mg/kg/day for 6 days, 0.75 mg/kg/day for 12 days, and no medication. Medicated feed was scheduled so that all treated pigs reached the last day of treatment on the same day, thus making the time between the last treatment and necropsy equal for all groups. Ascaris suum and Trichuris suis were the target species, their presence before treatment being determined by fecal egg counts and at necropsy by worm counts. At necropsy, 9 control pigs were infected with A suum (mean of 18.0 worms/pig), and all control pigs had T suis infection (mean of 36.5 worms/pig). All 3 treatment schedules were 100% effective in removal of A suum; and for T suis, the 3-day regimen was 100% effective, the 6-day regimen, 99.2%, and the 12-day regimen, 91.0%.     

Determination of anthelmintic efficacy against equine cyathostomins and Parascaris equorum in France

This article reports the results of a faecal egg count reduction test on 4 farms in France, as an integrated part of the routine deworming strategy against horse cyathostomins and Parascaris equorum. Treatment with fenbendazole (FBZ) or ivermectin (IVM) was evaluated in yearlings on Farms 1 and 2 and treatment with pyrantel embonate (PYR) was tested on Farms 3 and 4. Calculation of the arithmetic mean faecal egg count reduction and the 95% confidence intervals (95% CI) around the mean was performed using bootstrap analysis. For equine cyathostomins, resistance to FBZ was found with an arithmetic mean reduction of 48.8% (95% CI: 1.9–69.3%). On Farms 1 and 2, horses with reduced efficacy were identified. PYR was found to be effective against cyathostomins, with an arithmetic mean reduction of 95.3% (95% CI: 84.6–99.8%), as well as IVM (100%). For P. equorum, both FBZ and PYR were effective (100% reduction). The efficacy of IVM, however, was low (45.5%; 95% CI: 0–96.3%). These results confirm that FBZ resistance in equine cyathostomins is present in France and that anthelmintic resistance to IVM is present in P. equorum. This study underlines the necessity to evaluate the efficacy of horse deworming strategies on a regular basis under field conditions.     

Acute toxicity of paraherquamide and its potential as an anthelmintic.

Paraherquamide, an oxindole alkaloid metabolite of Penicillium paraherquei and P charlesii, is a new anthelmintic with potential broad-spectrum use. In initial trials, it had an excellent safety profile in cattle and sheep at doses efficacious against a dozen or more helminths, but recently it produced unexpected and severe toxicosis in dogs at doses far below those that were safe in the ruminants. To provide data on which to build rational safety tests in the future, we tested the acute toxicity of paraherquamide administered PO to male CD-1 mice and compared its profile with the most potent anthelmintic known, ivermectin. The estimated doses lethal to 50% of a group of mice were 14.9 and 29.5 mg/kg of body weight for paraherquamide and ivermectin, respectively. The no-effect doses were 5.6 and 18.0 mg/kg for paraherquamide and ivermectin, respectively. Signs of intoxication in paraherquamide-treated mice, if they developed, emanated within 30 minutes of administration, irrespective of dose, and consisted of either mild depression with complete recovery or a 5- to 10-minute period of breathing difficulty followed by respiratory failure and death by 1 hour after treatment. Gross necropsy findings in paraherquamide-treated mice that died in the high-dose group were normal. Ivermectin-related toxicity was slower and more predictable, taking place over a 3-day period, with dose-dependent signs of intoxication consisting of tremors, ataxia, recumbency, coma, and death. Necropsy of ivermectin-treated mice that died in the high-dose group revealed dehydration, a condition most likely resulting from the coma-induced state.(ABSTRACT TRUNCATED AT 250 WORDS)