Effect of a high dosage of ketoconazole all or not combined with ovariectomy on N-nitroso-N-methylurea-induced mammary cancer in the rat.

Rats bearing mammary carcinomas induced by N-nitroso-N-methylurea (NMU) were subjected to either no treatment (C), to ovariectomy (O), to continuous ketoconazole dosing at 400 ppm into the diet (K) or to both ovariectomy and ketoconazole dosing (O + K). In both C and K groups survival over the experimental period of 15 weeks was low (12.5% and 15.8%, respectively). In the O group and the O + K group survival markedly improved (52.6%, P less than 0.05 and 68.4%, P less than 0.01, respectively) and partial or complete tumor regression was noted respectively in 2 and 8 rats out of 19. Whereas C and K dosed groups showed an equally high tumor multiplicity and fast tumor growth, both parameters were markedly reduced in the O group and more markedly reduced in the O + K group which showed an overall decrease of the number of tumors. These findings indicate a synergistic effect of ketoconazole dosing and ovariectomy.

In vivo mutagenicity evaluation of domperidone in Drosophila germ cells and rat bone marrow cells.

Possible induction of chromosome aberrations and gene mutations by domperidone was studied in vivo respectively by a micronucleus test on female rats and a sex-linked recessive lethal test on Drosophila. In accordance with previous results all these studies revealed negative findings for domperidone so that it can be concluded that domperidone has no potential to induce chromosome aberrations and/or gene mutations.

Sampling times in micronucleus testing.

A series of micronucleus inducers were evaluated in the mouse bone marrow micronucleus test to determine if a 72-h sampling time enhances the sensitivity for detecting genotoxic agents. Male and female Swiss albino mice were dosed once with 7,12- dimethylbenz[a]anthracene, 6-mercaptopurine, benzo[a]pyrene, benzene, cyclophosphamide, 2-acetylaminofluorene, tubulazole, or mitomycin C. According to the EEC and OECD guidelines, the mice were killed at 24, 48 and 72 h after dosing. All test compounds induced an increase in the number of micronucleated polychromatic erythrocytes at 24 and/or 48 h. From the results obtained, it was evident that the 72-h sampling time does not enhance the sensitivity of the micronucleus test. The present data show that for screening purposes two sampling times at 24 and 48 h are sufficient to detect clastogens as well as aneugens. Although quantitative differences were found in sensitivity to micronucleus inducers between male and female mice, no qualitative differences were observed between the two sexes.

Genetic toxicology in the pharmaceutical industry.

The main objective of the pharmaceutical industry is health protection. Drugs not intended for use in life-threatening diseases should be free from toxic effects, but every natural or man-made chemical has potential toxicity depending on the exposure dose. The pharmaceutical industry considers genetic toxicology as a part of overall safety evaluation. No single genetic toxicity test is satisfactory; a battery of test is necessary to cover the whole spectrum of genetic events. Numerous tier approaches have been proposed for mutagenicity testing but from the toxicological viewpoint a phylogenic testing model seems more appropriate than a sequential step model. Evaluation of the mutagenic potential of drugs should rely on in vivo animal models, although for screening purposes and to promote understanding of the mutagenic action, in vitro tests using different systems can be used. Rejection solely on the basis of in vitro tests can lead to the unnecessary loss of a valuable drug. More inexpensive and relatively short tests on non-mammalian and mammalian cells are needed for studying structure-mutagenic activity relationships. For extrapolating to man and in the framework of clinical studies, it might be worthwhile to focus more time on developing inexpensive and simple tests using models directly relevant to man (e.g. human body fluids).

Efficacy of diclazuril in the control of intestinal coccidiosis in rabbits.

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.

Efficacy of diclazuril in the prevention and cure of intestinal and hepatic coccidiosis in rabbits.

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.

Efficacy of diclazuril against Eimeria dispersa in turkeys.

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.

Efficacy of diclazuril against turkey coccidiosis in a floor-pen experiment.

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.

Efficacy of diclazuril against turkey coccidiosis in dose-titration studies.

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.

In vivo action of the anticoccidial diclazuril (Clinacox) on the developmental stages of Eimeria tenella: a histological study.

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.

In vivo action of the anticoccidial diclazuril (Clinacox) on the developmental stages of Eimeria tenella: an ultrastructural evaluation.

A single 5-mg/kg oral dose of diclazuril affected both the asexual and sexual development of Eimeria tenella in experimentally inoculated chickens. In second-generation schizonts, early growth and nuclear divisions progressed normally, but a marked inhibition of merozoite formation was observed. Exogenesis of merozoites was largely prevented, whereas production of micronemes, amylopectin granules, and dense bodies and the formation of rhoptries, conoid, and pellicle continued. All these subcellular organelles accumulated, together with differentiated nuclei, within the main cytoplasmic mass. In the end, complete necrosis of the schizonts occurred. In macrogamonts, dilation of the rough endoplasmic reticulum around type II wall-forming bodies, fusion of type II wall-forming body contents, disturbance of the normal parallel arrangement of rough endoplasmic reticulum, and disruption of row formation of amylopectin granules became evident. In the microgamonts, normal evagination of microgametes was prevented; the flagellar complex formed within the main cytoplasmic mass and the differentiated nuclei remained present within the parasite body. The macro- and microgamonts also ended up in a stage of complete necrosis. These data indicate that diclazuril treatment primarily affects the normal differentiation of the respective endogenous stages during parasite development. This leads to complete degeneration of schizonts and gamonts indicating the lethal effect of this new anticoccidial compound.

Action of the anticoccidial clazuril on the endogenous stages of Eimeria labbeana and E columbarum in experimentally infected pigeons.

Racing pigeons were artificially infected with a mixed inoculum of Eimeria labbeana (85 per cent) and E columbarum (15 per cent) and treated orally with 2.5 mg clazuril either on day 1, 2, 3, 4, 5, 6 or 7 after infection. The impact of the treatment on the different developmental stages was evaluated by oocyst output and by histological examination of the duodenum and jejunum. The life cycle always became completely interrupted, but maximal effects were noted when treatment was given on day 4, 5 or 6 after infection. Treatment during patency completely interrupted oocyst excretion within three days after dosing. Degenerative changes in schizonts and gametocytes were always observed. The histology revealed a reduced number and abnormal structure of developing merozoites; a ballooned aspect and presence of numerous small vacuoles in the microgametocytes; the absence of typical wall-forming bodies in macrogametocytes and a complete absence of oocysts. It is concluded that clazuril has a coccidiocidal effect on the asexual and sexual developmental stages of both Eimeria species, resulting in a complete interruption of the life cycle.

Safety studies evaluating the effect of mebendazole on liver function in dogs.

The effect of repeated and exaggerated mebendazole administration was evaluated in dogs with and without experimentally induced altered liver function. Irish Setters and Toy Poodles were dosed at 1, 3, and 5 times the therapeutic dose (22 mg/kg) of mebendazole for 17 days, without any effect on the liver. Mixed breed dogs that received increasing doses of mebendazole at 11 to 110 times the therapeutic dose for 2 months did not show adverse effects and remained in good health throughout the experiment. There was not substantial evidence that carbon tetrachloride-induced liver changes were exacerbated by subsequent repeated treatments with mebendazole at 15 times the therapeutic dose. Additionally, in dogs whose liver function was compromised experimentally by glutathione depletion and microsomal enzyme induction, administration of mebendazole at this same dosage for 30 days did not result in any hepatotoxic effect. When mebendazole was given at 15 times the therapeutic dose prior to an hypoxic episode in dogs pretreated with a barbiturate and high protein diet, there was no evidence of any adverse effect on liver function. These metabolic manipulations, in conjunction with mebendazole administration, failed to reveal any mechanism of hepatic dysfunction associated with mebendazole treatment. Unrecognized factors appear to be involved with the rare cases of hepatic dysfunction that have been reported after mebendazole administration. Only careful documentation of field cases and further laboratory research can identify these factors.

Effect of diclazuril (Clinacox) on the development of protective immunity against Eimeria tenella: laboratory trial in broiler chickens.

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.

Diclazuril, a new broad spectrum anticoccidial drug in chickens. 1. Dose titration studies and pilot floor pen trials.

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.

Diclazuril, a new broad spectrum anticoccidial drug in chickens. 2. Battery trials.

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.

Diclazuril, a new broad-spectrum anticoccidial for chickens. 3. Floor-pen trials.

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.

Efficacy of flubendazole against gastrointestinal and lung nematodes in pigs.

Three trials were carried out on landrace pigs of various ages to assess the anthelmintic efficacy of flubendazole. The pigs were either artificially infected with Metastrongylus apri or naturally or artificially infected with the gastrointestinal nematodes Ascaris suum, Oesophagostomum dentatum or Hyostrongylus rubidus. For mass medication of young pigs and fatteners a dose regimen of 30 ppm flubendazole in the feed for 10 consecutive days was 100 per cent effective against the four nematode species. For individual medication a single dose of 5 mg/kg bodyweight administered in a small amount of feed was also 100 per cent effective. No side effects were observed.

Anticoccidial efficacy of diclazuril in pheasants.

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.

[Safety of a new anticoccidial agent, clazuril, during reproduction in carrier pigeons]. / Veiligheid van een nieuw anti-coccidiosemiddel clazuril tijdens de kweekperiode bij postduiven.

In a six-month study, the safety of repeated treatments with clazuril, a new anticoccidial agent for the treatment of pigeons, was evaluated during 4 to 5 consecutive reproduction cycles. Clazuril was administered weekly at the recommended therapeutic dose level of 2.5 mg/pigeon and at twice this dose. The results were compared to placebo treatment. Evaluation was based on the following parameters: reproductive behaviour, egg laying, fertilisation, embryonic mortality, hatching, percentage of weaning and time interval between consecutive reproductive cycles. Maternal-paternal toxicity (body weight evolution, feather coat) and tolerance in newborn pigeons (general behaviour, daily gain in body weight, feathering, skeletal anomalies) were also evaluated. In all the treatment groups, reproductive performance was comparable and no drug- or dose-related side-effects were observed. Fertilisation, embryonic mortality and the time-interval between the consecutive reproductive cycles remained within the normal physiological range. Hatched pigeons showed a satisfactory daily weight gain and deaths among these birds were exceptional. Tolerance of clazuril in adult as well as in day-old pigeons was exceptionally good and body weight evolution, feathering and mortality were not adversely affected. No drug-related pathological findings or skeletal anomalies were detected in the animals at autopsy. It can be concluded that repeated treatments with clazuril are entirely harmless for pigeons during reproduction, even at double the therapeutic dose level. Clazuril is well tolerated by newborn and growing pigeons, which means that pigeon house group treatments during reproduction may be performed without any risk.