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Guidelines have been designed to assist in the planning, operation and interpretation of studies for the assessment of the efficacy of drugs against helminth parasites of dogs and cats. The advantages, disadvantages and application of critical and controlled tests are presented. Information is also provided on the selection of animals, housing, feeding, dose-titration, confirmatory and clinical trials, record keeping and necropsy procedures. These guidelines should assist both investigators and registration authorities involved in the evaluation of anthelmintics to employ comparable and standard procedures and will have the added benefit of minimising the numbers of animals needed for such tests.

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Diclazuril, a new anticoccidial drug, was tested for its efficacy against Eimeria dispersa in turkeys. A dose-titration study indicated that diclazuril at dosages of 0.5, 1, and 2 ppm in the feed was highly effective in terms of weight gain and suppression of lesions, abnormal droppings, and oocyst shedding.

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The effect of diclazuril, fed to chickens at 1 ppm in the feed, was studied to determine whether the drug interfered with the development of immunity to Eimeria tenella. Group A was not treated, Groups B and C received diclazuril from Day 1 until Day 15, after which time the medicated feed was replaced by blank feed for the remainder of the experiment from Day 16 until Day 42. Immunization was performed in Groups A and B by artificial trickle infections with 2,000 sporulated oocysts per bird on Days 4, 6, 8, 11, and 13. On Day 29, a challenge infection was given using 200,000 oocysts per bird. The unmedicated birds (Group A) developed subclinical coccidiosis after the trickle infections with excretion of oocysts and a slightly decreased growth performance. At challenge, a good protective unimmunity was present, reflected by a good growth performance and a low oocyst excretion. The unimmunized birds (Group C) developed a severe clinical disease after challenge with high oocyst output, increased mortality, and poor growth performance. The diclazuril-medicated, trickle-infected birds (Group B) were well protected, both against the immunizing trickle infections and the challenge infection. After challenge, no clinical disease developed, although some lesions and oocyst excretion were present. It is concluded that, under the conditions of the trial, diclazuril did not significantly interfere with protective immunity formation against E. tenella.

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The efficacy of levamisole pour-on against Dictyocaulus viviparus was compared to that of subcutaneous levamisole injection. Eighteen calves were raised individually and artifically infected with D. viviparus larvae. Faecal samples were collected 27 and 28 days later and larvae per gram (l.p.g.) determined. The animals were then divided into three comparable groups. Group 1 animals remained untreated as controls. Group 2 animals received levamisole 10% w/v subcutaneous injection at a dose of 5 mg kg-1 and Group 3 received levamisole pour-on 20% w/v at a rate of 10 mg kg-1 applied transdermally. Results of l.p.g. measurements from faecal samples taken 7 and 8 days post-treatment indicated a dramatic reduction in the worm burden of animals in both treatment groups. Necropsies at 14 days post-treatment revealed few adult worms in these groups, indicating a 99 and 98% kill rate for pouron and subcutaneous injection, respectively.

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This study investigates the level of helminthic and protozoal infestation over the last 10 years in strays, well-cared-for dogs and cats. Determination of the prevalence of infections was based either on faecal examination or on worm counts at necropsy. Of 2324 faecal flotations (NaCl sp.gr. 1.20) of stray dogs, 34.2% had eggs or proglottids of one or more worm species consisting of Toxocara canis (17.4%), Toxascaris leonina (10.1%), Uncinaria stenocephala (11.4%), Trichuris vulpis (7.0%) and cestodes (2.1%). Isospora oocysts were observed in 5.2% of the dogs. The data on the distribution of the various worm species in the positive dogs indicate that T. canis eggs were by far the most common (50.9%). Necropsy data from 212 infected dogs indicate that 38.9% were infected with T. canis and 33.7% with T. leonina. The overall prevalence of worm infestation of 246 well-cared-for kennel dogs, based on worm egg counts by the McMaster technique, was 36.1%. Of 30 feline faecal samples examined by flotation, 83.3% were positive for parasites, including Toxocara cati (60%), Ancylostoma tubaeformae (36.6%), Taenia (Hydatigera) taeniaeformis (20%) and coccidia (30%). Toxocara cati was the most frequently found worm species at the necropsy of 25 cats (52%). Toxoplasma was not observed.

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The effect of closantel (10 mg/kg orally) and triclabendazole (10 mg/kg orally) on the reappearance of a patent infection of Fasciola hepatica was studied in experimentally infected sheep. The treatments resulted in the interruption of faecal egg output for 11 weeks with triclabendazole and 13 weeks with closantel. Necropsy of untreated control animals revealed a mean burden of 360 flukes with a mean (+/- se) surface area of 171 +/- 64.3 mm2, whereas the fluke burdens in the closantel and triclabendazole-treated animals 14 weeks after treatment were 61 (83 per cent reduction) and 21 (94 per cent reduction), respectively. The surface areas of the flukes in the triclabendazole-group were comparable with the untreated controls (141 +/- 51.8 mm2), but the flukes in the closantel group were markedly smaller (43.1 +/- 26.9 mm2). It is concluded that closantel has, in epidemiological terms, a potency comparable with that of triclabendazole, despite its slightly lower efficacy against the very immature stages.

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Diclazuril, a new anticoccidial drug, was tested in young pheasants artificially infected with the three most common pathogenic species of Eimeria, E colchici, E duodenalis and E phasiani. In two replicate experiments each with 40 birds the mortalities in the infected controls were 50 and 25 per cent. Diclazuril was administered in the feed at dose levels of 1, 2 and 4 ppm from the day before the inoculation of coccidia until the end of the test on day 6 after infection. The 1 ppm dose failed to inhibit the development of the parasite completely, as was shown by a reduction of the weight gain of the birds and the output of a small number of oocysts. Diclazuril at 2 or 4 ppm adequately controlled the infection, with weight gains similar to those of the uninfected controls. At all dose levels, mortality, intestinal lesions and diarrhoea were prevented.

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Diclazuril is a benzeneacetonitrile showing great promise as a broad-spectrum anticoccidial agent for chickens, turkeys, and rabbits. The high anticoccidial activity of diclazuril in chickens, as first reported in dose-titration studies and battery trials, was confirmed in three floor-pen trials. The efficacy was demonstrated against six major pathogenic species of Eimeria after artificial infection with one or more species. The experimental data indicated that diclazuril, at dose levels of .5, .75, 1, and 2 ppm, had a high anticoccidial activity in terms of preventing mortality, suppressing or reducing lesion scores, and allowing for normal weight gains as well as productivity. The performances obtained with diclazuril was generally comparable with that of salinomycin at 60 ppm and that of lasalocid at 90 ppm.

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A total of 50 jirds (Meriones unguiculatus) were infected by intraperitoneal implantation of 0.20 g metacestode tissue of Echinococcus multilocularis. The feed of 4 groups of 10 animals each was treated for 35 days with 500 ppm albendazole, flubendazole, mebendazole or praziquantel; 1 group of 10 jirds served as unmedicated controls. At autopsy 2 weeks after the end of medication, the transplants had increased in size and weight, reaching a total weight of 78.11 g in the control group, 21.60 g in the albendazole-treated group, 3.63 g in the flubendazole-treated group, 7.00 g in the mebendazole-treated group and 68.91 g in the praziquantel-treated group. The percentage in reduction of parasitic tissue weight as compared with control values was 72%, 95%, 91% and 12%, respectively. The calculated daily drug intake was approximately 48 mg/kg body weight. Drug-related side effects were not observed.

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The anticoccidial efficacy of diclazuril was studied in rabbits artificially infected with Eimeria flavescens, Eimeria intestinalis, Eimeria magna and Eimeria perforans. Continuous administration at 1 and 2 ppm in pelleted feed proved to be highly efficacious in controlling oocyst output and faecal scores. The weight gain was comparable and the feed efficiency slightly improved compared with the non-infected, non-medicated controls, and clinical signs were fully prevented. Medication of rabbits at 0.5 ppm also provided a significant improvement in all parameters compared with the infected, non-medicated controls. In order to obtain 100% effectiveness in the control of intestinal coccidiosis in rabbits, continuous medication at 1 ppm is recommended.

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The efficacy of diclazuril against intestinal and hepatic coccidiosis was studied in artificially infected rabbits. Prophylaxis against intestinal coccidiosis was evaluated using a mixed infection of Eimeria intestinalis, Eimeria magna and Eimeria perforans. Continuous medication in the feed at 1 p.p.m. was 100% effective in reducing oocyst output and faecal scores, and weight gain and feed efficiency were normal. Hepatic coccidiosis induced by Eimeria stiedai was prevented at 0.5 and 1 p.p.m. as shown by negative oocyst counts, normal liver weight, absence of liver lesions, and normal body-weight gain and feed efficiency. Medication at 1 p.p.m. for 7 consecutive days during the prepatent phase of hepatic coccidiosis resulted in large reductions in oocyst counts and lesion scores with a normal liver weight and growth performance. Diclazuril at 1 p.p.m. in the feed prevented both intestinal and hepatic coccidiosis in rabbits and can be advocated for safe mass medication.

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Diclazuril, a new anticoccidial drug, was tested for its efficacy in turkeys against single Eimeria infections. Dose-titration studies indicated that diclazuril at dosages of 0.1, 0.5, 1, and 2 ppm was highly effective against the major pathogenic species-E. adenoeides, E. gallopavonis, and E. meleagrimitis-in terms of weight gain and suppression of lesions, abnormal droppings, and oocyst shedding.

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Diclazuril, a new anticoccidial drug, was tested for its efficacy in turkeys against mixed Eimeria infections. A floor-pen trial indicated that diclazuril at dosages of 0.5 ppm and 1 ppm in the feed was highly effective against the major pathogenic species E. adenoeides, E. gallopavonis, and E. meleagrimitis in suppressing intestinal and cecal lesions and oocyst shedding. Weight gain and feed conversion improved, particularly at 1 ppm.

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Diclazuril, a new anticoccidial drug, was tested in poultry against six Eimeria species either in single species infections in dose titration studies or in mixed species infections in floor pen trials. The dose titration studies in cockerels indicated that diclazuril at dosages of 10, 5, 1, and .5 ppm was highly active against all major pathogenic species: E. tenella, E. acervulina, E. necatrix, E. brunetti, E. maxima, and E. mitis and effective in terms of weight gain and suppression of mortality, dropping scores, and oocyst counts. In two floor pen trials diclazuril was fed for 6 wk to broiler chickens experimentally infected either with E. tenella and E. acervulina or with E. necatrix and E. brunetti. Dose levels of 10, 5, and 1 ppm suppressed mortality and lesion scores. Even at 1 ppm the mean terminal body weight, feed conversion, and productivity index of treated birds were comparable to results for the uninfected, unmediated controls. In these pilot studies, involving 1,020 Hisex and 1,000 Hubbard broiler chickens, it has been demonstrated that diclazuril at the dose level of 1 ppm in the diet is an excellent anticoccidial without any adverse effects.

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Battery trials have confirmed the broad spectrum anticoccidial activity of diclazuril as previously reported in dose titration studies. The advocated dose level of 1 ppm in the diet demonstrated excellent activity against the economically most important Eimeria species. At this dose level, body weight gains were comparable to those of uninfected, unmedicated controls and the oocyst production was negative in most species. Lesion scores and dropping scores were nil or highly reduced. An E. maxima-147 strain, less sensitive to ionophores, also responded well to diclazuril. It was concluded that diclazuril is a promising anticoccidial for the control of all species of coccidia that cause losses to the poultry industry.

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The ultrastructural morphology of the different endogenous stages of Eimeria maxima and E. brunetti was evaluated after oral treatment of inoculated chickens with a single dose of 5 mg/kg diclazuril. The drug induced no ultrastructural change in the growth or differentiation of the various schizont stages of both Eimeria spp. In E. maxima, the micromorphological appearance of micro- and macrogamonts developing from the blast from to maturation also remained unaffected by drug treatment. However, in all fertilized macrogamonts the normal pattern of oocyst wall establishment was completely disturbed, resulting in the formation of an abnormally thickened, incomplete oocyst wall and the necrosis of the zygote. In E. brunetti, the growth and nuclear division during microgametogenesis were not affected but differentiation was clearly abnormal. In comparison with the controls, this abnormal differentiation was characterized by a less extensive enlargement of the parasite surface area, aberrant morphological configurations of condensed heterochromatin, intracytoplasmic flagella formation, and glycogen accumulation. Finally, the complete degeneration of all microgamonts ensued. The growth and differentiation leading to mature macrogamonts was not disturbed; however, subsequent oocyst wall formation was largely precluded and the macrogamonts proceeded to degenerate completely. We conclude that diclazuril treatment primarily affected particular stages in the sexual development of both Eimeria spp., resulting in the complete eradication of these coccidian species.

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Diclazuril, a new benzeneacetonitrile anticoccidial, has potent activity against various stages of Eimeria tenella. A single treatment of experimentally infected chickens during the prepatent phase (up to day 5) results in a complete interruption of the life cycle and oocyst shedding. The first- and second-generation schizonts show extensive degenerative changes that finally result in a complete loss of the parasitic stage. The degeneration is characterized by loss of internal structure, the appearance of many intracytoplasmic vacuoles, and incomplete merogony. The merozoites themselves show similar degenerative changes, including the presence of numerous small vacuoles in the cytoplasm. Diclazuril is also effective against both the micro- and macrogametocytes that have a ballooned appearance and loose their internal structure completely. In the macrogametocytes, wall-forming bodies either do not develop or disappear rapidly. Development of typical caecal lesions is prevented when treatment with diclazuril is initiated before large numbers of second-generation schizonts appear, i.e., day 3. It is concluded that diclazuril is lethal against both the asexual and the sexual stages of E. tenella. At the proposed use level of 1 ppm in the feed, the life cycle is interrupted at a very early stage and lesion development and oocyst shedding are completely prevented.

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