ANTHELMINTIC PREPARATIONS

Abstract: A veterinary preparation comprising an effective amount of an anthelmintic agent, preferably fenbendazole, and a non-­ toxic amount of selenium salt, preferably sodium selenate, ef­ fective to prevent selenium deficiency in an animal. Preferably, a non-toxic amount of a cobalt salt effective to prevent cobalt deficiency in an animal is also included. By combining selenium with an anthelmintic agent, the safety margin for selenium is greatly increased.

Description: Anthelmintic preparations The present invention relates to anthelmintic preparations, particularly for the treatment of ruminants. As clinical and sub-clinical nematode infections can cause serious economic loss, due to death, reduced weight gain and lowered milk yield, anthelmintic treatment of ruminants is now a routine practice. Treatment is also undertaken to reduce pasture larval contamination and thus aids in ensuring optimal grassland usage. In some cases, however, even after anthelmintic treatment, animals still fail to thrive. It is now considered that this may be caused by mineral deficiencies resulting from soils and pastures deficient in minerals. Cobalt is one mineral that may be lacking in the diet of ruminants, particularly in the spring, when there is pasture dominance of rapidly growing grasses, which have a lower cobalt content than more slowly growing grasses. Cobalt participates in the production of vitamin B12 in the rumen, and deficiency results in loss of appetite and loss of bodyweight, and the animal may finally waste away. In the ewe, in addition to weight loss, milk yield falls and there is a reduction in fleece quality. Selenium is also an essential element, but its exact metabolic function is uncertain. Deficiency is associated with muscular dystrophy, unthriftiness and infertility in female animals. It has, therefore, been proposed to administer trace elements, particularly cobalt and selenium, to animals. Some of the trace elements, however, are toxic, and it is possible to overdose animals. This is particularly so with selenium, the oral LD50 of which is only 0.5 mg/kg. The present invention is based on the surprising observation that a preparation comprising both selenium and an anthelmintic agent has a very greatly increased safety margin with regard to the selenium. Accordingly, the present invention provides a veterinary preparation which comprises an anthelmintic agent and a selenium salt, preferably in admixture or conjunction with a carrier suitable for use in a veterinary preparation. Preferably, the preparation also comprises a cobalt salt. The anthelmintic agent may be suitable for treating worms and/or flukes, and is preferably a benzimidazole anthelmintic, for example, parbendazole, oxfendazole, oxibendazole, thiabendazole, albendazole, cambendazole, mebendazole, flubendazole or fenbendazole. As a general rule, the less soluble a benzimidazole anthelmintic, the more active it is. Accordingly, fenbendazole, oxfendazole and albendazole are more active than thiabendazole, parbendazole and cambendazole and are, therefore, preferred. Fenbendazole is a broad spectrum anthelmintic with excellent efficacy against adult, larval and egg stages of all important gastro-intestinal and respiratory nematodes of ruminants. In addition, it is effective against both scolices and segments of M. expansa, the cestode affecting sheep and cattle. It is non-teratogenic, and so can be used safely in animals at any stage of pregnancy. Having no adverse effect on fertility, it can be used around the time of service and in the breeding male. It has a very high safety margin (sheep: 1000 times the recommended dose, cattle: 50 times the recommended dose). Moreover, it is non-toxic to handlers. Oxfendazole and albendazole have substantially the same spectrum of activity as fenbendazole, but have a lower therapeutic index. Fenbendazole (methyl-5phenylthio-2-benzimidazole carbamate) is particularly preferred as the anthelmintic agent for the preparation of the present invention. A preparation of the invention may be in liquid or solid form. A liquid preparation may be a solution, but is generally a suspension, preferably a fairly mobile suspension. Other forms of preparation suitable for dosing animals may be used, for example, tablets, bolus tablets and pastes; and powders and granules for reconstitution as solutions or suspensions prior to use. Moreover, animals may be dosed by the addition of an anthelmintic agent, a selenium salt and, preferably also a cobalt salt, directly to the feed. A preparation of the invention, accordingly, may be in a form suitable for addition to an animal feedstuff. The invention further comprises an animal feedstuff that has been augmented by the incorporation of an anthelmintic agent, a selenium salt and, preferably, also a cobalt salt. In a preparation of the invention, the selenium salt may be insoluble or soluble, soluble salts being preferred, for example, alkali metal and ammonium salts. The selenium may be in the form of a selenate, selenite or selenide. Examples of selenium salts are sodium, potassium and ammonium selenate, sodium, potassium and ammonium selenite, and sodium and potassium selenide. Selenates are generally preferred as they are the most soluble, and sodium selenate is particularly preferred. The cobalt salt also may be insoluble or soluble, examples of cobalt salts being cobalt carbonate, cobalt chloride, cobalt nitrate and cobalt sulphate. Cobaltous or cobaltic salts may be used Soluble cobalt salts are preferred. In a liquid preparation, the selenium salt and, when present, the cobalt salt, is preferably in solution to ensure a uniform distribution of these elements throughout the preparation and therefore to improve dose to dose reproducibility. One or more further physiologically active substance(s) may be present in a preparation of the invention, for example, selected from vitamins and, especially, other trace elements, for example, copper, zinc and magnesium. One or more carriers suitable for veterinary preparations may be present in addition to the active substances to aid manufacture and/or to confer desirable properties on the final preparation. Such materials include pH adjusters, pH buffers, wetting agents, suspending agents, thickening agents, stabilising agents, solubiZisers, antimicrobial preservatives, lubricants and disintegration agents. Water is generally the basis for a solution or suspension. A suspension of the invention may be prepared by mixing the ingredients with water using a high speed mixer/ homogeniser until a uniform dispersion is achieved. The pH of the suspension is usually adjusted to the range 5.5 to 6.5 and especially to 5.9 to 6.0 to optimise product stability. The suspension preferably includes ingredients to ensure an elegant product by retarding sedimentation and/or aiding resuspension on shaking, for example, a thixotropic agent, eg. colloidal silicon dioxide, and/or a thickening agent, eg. carboxymethyl cellulose. The selenium salt is preferably added as a solution to avoid dust problems on handling. Other preparation forms, for example, tablets,powders and granules, may be prepared in a conventional manner. In a suspension of the invention the fenbendazole content is, for example, in the range of from 10 to 40 mg/ml for sheep and goats,especially 25 mg/ml. For cattle, the fenbendazole content is, for example, from 20 to 200 mg/ml, especially 100 mg/ml. If a different anthelmintic agent is used, the dose should be substantially equivalent to that given above for fenbendazole. (The recommended doses of most anthelmintic agents for sheep,goats and cattle are generally well known.) The content of selenium in a suspension of the invention, calculated as elemental selenium is, for example, within the range of from 0.1 to 2.0 mg/ml, especially 0.4 mg/ml for sheep and goats, and within the range of for from 0.1 to 2.5 mg/ml, especially 2.0 mg/ml for cattle. The cobalt content, also calculated as the elemental substance is, for example, from 0.1 to 5.0 mg/mlF especially from 0.9 to 1.0 mg/ml for sheep and goats, and from 0.1 to 5.0 mg/ml,especially 2.5 mg/ml for cattle. If desired, however, the selenium content can be increased considerably, for example, up to 10 mg/ml, for sheep, goats and cattle. A suspension of the invention for sheep and goats preferably comprises 25 mg of fenbendazole, 0.4 mg of elemental selenium and 0.95 mg of cobalt per ml. With this preparation, the recommended dose is 1 ml/5 kg body weight. A suspension of the invention for cattle preferably comprises 100 mg of fenbendazole, 2.0 mg of elemental selenium and 2.5 mg of elemental cobalt per ml. The recommended dose of this preparation is 7.5 ml per 100 kg body weight. In a tablet or bolus tablet of the invention, the doeses of the various ingredients may be calculated pro rata from the doses given above, knowing the body weight of the intended recipient of the dose, for example, in a tablet suitable for administration to a 50 kg sheep, the fenbendazole content is generally from 100 to 400 mg, especially from 250 to 300 mg, the content of elemental selenium is generally from 1 to 20 mg, especially from 3 to 5 mg; and the cobalt content, calculated as elemental cobalt, is generally from 1 to 50 mg, especially from 9 to 10 mg. Tablets may be scored to assist the adminis tration of divided doses to smaller animals, and multiple doses may be given to larger animals. The optimum dose of the various ingredients for solid preparations e.g. tablets and bolus tablets for cattle can be calculated similarly from the data given above for suspensions. As indicated above, appropriate amounts of other anthelmintic agents can be readily calculated. It will be appreciated that as an alternative to presenting the anthelmintic agent and the selenium to an animal in a single preparation, these two components may be formulated separately and administered simultaneously or substantially simultaneously. The present invention accordingly also provides a pack which comprises an anthelmintic agent and, in conjunction or close juxtaposition therewith, a selenium salt. The anthelmintic agent is in the form of a veterinary preparation and is preferably in admixture or conjunction with a carrier suitable for use in a veterinary preparation. The preparation may be liquid or solid, and example of preparation forms are given above. As indicated above, suspensions are generally preferred. The content of the anthelmintic agent is preferably as indicated above. The selenium salt may be formulated either alone or in admixture or conjunction with a carrier suitable for veterinary use in solid or liquid form, for example, as tablet, bolus tablet or paste, as a solution or suspension, or as a powder or granules for reconstitution as a solution or suspension prior to use. It is advantageous to provide th selenium in a unit dose form suitable for addition either to the suspension containing the anthelmintic agent or to water, preferably purified water. Tablets and sachets containing the appropriate amount of a selenium salt are examples of suitable dosage forms. The amounts of selenium to be incorporated can be calculated from the data given above. The cobalt salt may be present either as a component of the anthelmintic preparation or as a component of the selenium preparation. Examples of both selenium salts and cobalt salts are given above. The anthelmintic agent-containing preparation and the selenium-containing preparation should be provided in conjunction with one another or in close juxtaposition to one another for example, the two should be provided close to hand. Preferably, the two components are joined together, one is attached to the other, or both are provided in the same container. The selenium containing preparation, for example, tablets or a sachet containing a powder or granules for reconstitution, may be physically attached to the anthelmintic preparation, for example, a pack or bottle of tablets, or a bottle of liquid preparation. The two components may be provided together in a container, for example, a box or carton. Both components may be solid, both may be liquid, or one may be solid and one may be liquid. As indicated above, liquid forms may be preferred for the anthelmintic preparation, and a pack may comprise a bottle of anthelmintic suspension together with a bottle of selenium-containing preparation or a solid selenium preparation, for example, tablets or one or more sachets of powder or granules for reconstitution. Other preparation forms may be provided analogously. The two preparations should be accompanied by in structions regarding their use. The instructions should indicate that the two preparations should be administered simultaneously or substantially simultaneously, with the selenium component preferably being administered immediately after the anthelmintic component. The instructions should indicate that the selenium component can be added to the anthelmintic component, if this is appropriate. The selenium component can be administered separately. The appropriate dose of both components should be indicated. It may be convenient to provide the selenium component in units suitable for reconstitution in a predetermined amount of either water or anthelmintic suspension. The user may then decide whether to add the selenium component to the anthelmintic preparation, or to make it up with water and use it separately. The recommended treatment times for sheep with a preparation of the invention are as follows: Ewes: in the period within 4-6 weeks of lambing. In addition, further treatments may be given approximately 4-6 weeks after lambing and prior to service - the former to assist in reduction of pasture larval contamination and the latter to improve bodily condition and, therefore, conception rate. Such additional treatments by providing cobalt and selenium will aid in body condition maintenance and improve fertility. Lambs: at 4-6 weeks of age and monthly thereafter to maintain optimal growth rate. Treatments may have to be more frequent in areas where Nematodirus infection occurs. Bought-in treat on arrival and thereafter as for ewes/ Sheep: lambs, but particularly (as with all sheep) when moving to clean pasture. Treatment of cattle could be carried out as follows: Calves in their first grazing season: in a set stocked system, dose at 3 and 6 weeks post turnout and again at housing. Alternatively, dose and move to clean pasture in mid-summer, and dose again at housing. Cattle in subsequent grazing seasons: dose prior to movement to clean pasture and at housing. Cows: dose prior to calving. As indicated above, we have found, surprisingly, that the safety margin for selenium administration is increased greatly when the selenium is combined with an anthelmintic agent, particularly fenbendazole. The LD50 of elemental selenium per os in sheep is considered to be 0.5 mg/kg (W.M. Allen, Institute for Research on Animal Diseases, Compton), whereas we have found that administration of oral dose levels of elemental selenium of up to 2.0 mg/kg were tolerated, no signs of drug toxicity being observed in any of the treated animals. This decrease in toxicity was quite unexpected, and we believe that there is a safety margin of at least 25 times the recommended dose of the preparation of the invention. This is of considerable practical importance, as there is risk of killing livestock by administering an overdose of selenium using trace element preparations currently available, whereas it is virtually impossible to administer a fatal dose of selenium using a preparation of the present invention. The following Examples illustrate the invention. The anthelmintic agents denoted by their generic names are as follows. Fenbendazol: 5-Phenyl-thio-2-benzimidazoecarbamic acid metlnylester Oxfendazol: 5-Phenylsulfinyl-2-benzimidazolecarbamic acid methylester Albendazol: 5-Propyl-thio-2-benzimidazolecarbamic acid methylester Oxibendazol: 5-Propoxy-2-benzimidazolecal-bamic acid methylester Parbendazol: 5-Butyl-2-benzimidazolecarbamic acid methylester Mebendazol: 5-Benzoyl-2-benzimidazolecarhamic acid methylester Flubendazol: 5- (4-Fluorobenzoyl) -2-benzimidazolecarbamic acid methylester Cambendazol: 2-(4-Thiazolyl)-5-benzimidazolecarbamic acid isopropylester Thiabendazol: 2- (4-Thiazolyl) -benzimidazole Example 1 Composition per ml Fenbenda zole 25.000 mg Colloidal silicon dioxide 30.000 mg Sodium methylhydroxybenzoate 1.994 mg Sodium propylhydroxybenzoate 0.216 mg Sodium carboxymethyl cellulose 14.506 mg Polyvinylpyrrolidone 19.342 mg Sodium citrate 31.590 mg Citric acid 31.350 mg Cobalt sulphate hepthydrate 4.500 mg Sodium selenate decahydrate 60 % w/w solution 3.167 mg Purified water to 1.0 ml Procedure A mixing vessel was charged with water that had been filtered through a 0.45 ssm filter. The following ingredients were added, with mixing to disperse them: citric acid, sodium citrate, cobalt sulphate, sodium selenate solution, sodium methylhydroxybenzoate, sodium propylhydroxybenzoate and colloidal silicon dioxide. The following ingredients were added via an in-line homogenisation unit: sodium carboxymethyl cellulose, polyvinylpyrrolidone and fenbendazole. Circulation through the in-line homogeniser was continued until a uniform dispersion was achieved. The pH was checked and adjusted to 5.9 with citric acid if necessary. The resulting pink suspension was filled into 2 litre containers. This suspension contains 0.4 mg/ml elemental selenium and 0.94 mg/ml elemental cobalt. Example 2 Determination of efficacy and toxicity levels MATERIALS AND METHODS: Animals: Twenty, six to eight month old lambs were made available for the purpose of this investigation. The lambs were allocated to five groups (n = 4) on the basis of mean faecal egg counts determined on each of three successive days. Prior to treatment, all lambs were weighed. Lambs were maintained at grass as a single group for the duration of the study. TREATMENTS: Group 1: Untreated controls Group 2: Treated with 5 mg/kg fenbendazole as Panacur 2.5 % Suspension (Commercial batch, "Panacur", is a Trade Mark) Composition of Panacur 2.5 % : as described in Example 1 with the omission of the selenium and cobalt components. Group 3: Treated with 5 mg/kg fenbendazole as a develop mental formulation (see below) Group 4: Treated with 25 mg/kg fenbendazole as a developmental formulatIon (see below) Group 5: Treated with 50 mg/kg fenbendazole as a develop mental formulation (see below) All treatments were administered per os. DEVELOPMENTAL FORMULATION Procedure: As described in Example 1 Composition per ml: Fenbendazole 25.000 mg Colloidal silicon dioxide 30.000 mg Sodium methylhydroxybenzoate 1.994 mg Sodium propylhydroxybenzoate 0.216 mg Sodium carboxymethyl cellulose 14.506 mg Polyvinylpyrrol idone 19.542 mg Sodium citrate 31.590 mg Citric acid 31.350 mg Cobalt sulphate heptahydrate 4.500 mg Sodium Selenate decahydrate 4.700 mg Purified Water to 1.000 ml Groups 4 and 5 thus received five and ten times respectively the recommended dose of fenbendazole and cobalt, and 12.5 and 25 times respectively the recommended dose of selenium. Parasitological examinations: Faecal egg counts were performed using the modified McMaster technique on samples taken directly from the rectum of lambs pre-treatment and at seven, eleven, eighteen and twenty five days post treatment. Groups 1, 2 and 3 were slaughtered at eleven days post treatment and post-mortem examinations carried out. The abomasum, small intestine and large intestine of each animal were removed and the contents collected. The abomasal mucosa was digested in a pepsin/ hydrochloric acid mixture for six hours. Numbers of helminth species present were calculated from aliquot samples of the organ contents (abomasal digests being examined in toto). Clinical examinations: Clinical examinations were carried out on all lambs daily post treatment for the duration of the study. RESULTS: No signs of drug-associated toxicity were noted in any of the treated animals. Faecal egg counts of all lambs are given in Table 1. Treatment with the commercially available or experimental formulations virtually eliminated faecal egg excretions. Worm counts obtained from lambs in Groups 1-3 at slaughter are recorded in Table II. A mixed nematode burden was present in the untreated lambs - Haemonchus, Ostertagia and Trichostrongylus spp. being present in the abomasum; Cooperia, Nematodirus and Trichostrongylus spp. in the small intestine and Chabertia and Trichuris spp.in the large intestine. Treatment with both formulations virtually eliminated nematode burdens, there being no significant difference in efficacy between the two formulations. CONCLUSIONS: 1. Inclusion of the trace elements, cobalt and selenium, in Panacur 2.5 % Suspension had no deleterious effect on anthelmintic efficacy. 2. Oral dose levels of up to 1.9 mg/kg of elemental cobalt (as CoSo4. 71120) and 2.0 mg/kg of elemental selenium (as Na2SeO4#1OH2O) were tolerated, no signs of drug toxicity being noted in any of the treated lambs. EMI14.1 <tb> <SEP> Faecal <SEP> egg <SEP> count <SEP> (epg) <SEP> on <SEP> Treatment <SEP> day <tb> Group <SEP> Lamb <SEP> No. <SEP> Weight <SEP> (KG) <SEP> -3 <SEP> -2 <SEP> -1 <SEP> +7 <SEP> +11 <SEP> -18 <SEP> +25 <tb> <SEP> 844 <SEP> 32.0 <SEP> 1500 <SEP> 2400 <SEP> 1950 <SEP> 2150 <SEP> 2350 <tb> 1+ <SEP> 822 <SEP> 36.0 <SEP> 50 <SEP> 350 <SEP> 900 <SEP> 800 <SEP> 400 <tb> <SEP> 866 <SEP> 26.5 <SEP> 650 <SEP> 1900 <SEP> 350 <SEP> 650 <SEP> 400 <tb> <SEP> 812 <SEP> 32.0 <SEP> 50 <SEP> NIL <SEP> NIL <SEP> NIL <SEP> 50 <tb> <SEP> 870 <SEP> 35.0 <SEP> 250 <SEP> 350 <SEP> 650 <SEP> NIL <SEP> NIL <tb> 2+ <SEP> 823 <SEP> 33.0 <SEP> 700 <SEP> 350 <SEP> 1000 <SEP> NIL <SEP> NIL <tb> <SEP> 856 <SEP> 32.0 <SEP> 1500 <SEP> 1650 <SEP> 1300 <SEP> NIL <SEP> NIL <tb> <SEP> 880 <SEP> 38.0 <SEP> 1250 <SEP> 700 <SEP> 1100 <SEP> NIL <SEP> NIL <tb> <SEP> 851 <SEP> 27.0 <SEP> 1050 <SEP> 1100 <SEP> 1650 <SEP> NIL <SEP> 50 <tb> 3+ <SEP> 878 <SEP> 28.0 <SEP> 350 <SEP> 500 <SEP> 2000 <SEP> 50 <SEP> NIL <tb> <SEP> 826 <SEP> 32.5 <SEP> 750 <SEP> 600 <SEP> 750 <SEP> NIL <SEP> NIL <tb> <SEP> 2 <SEP> 28.5 <SEP> 650 <SEP> 650 <SEP> 1050 <SEP> NIL <SEP> NIL <tb> <SEP> 835 <SEP> 29.5 <SEP> 550 <SEP> 650 <SEP> 450 <SEP> NIL <SEP> NIL <SEP> NIL <SEP> NIL <tb> <SEP> 843 <SEP> 31.0 <SEP> 700 <SEP> 800 <SEP> 1650 <SEP> NIL <SEP> NIL <SEP> NIL <SEP> NIL <tb> 4 <SEP> 824 <SEP> 34.5 <SEP> 700 <SEP> 1000 <SEP> 1050 <SEP> NIL <SEP> NIL <SEP> NIL <SEP> NIL <tb> <SEP> 892 <SEP> 33.0 <SEP> NIL <SEP> NIL <SEP> NIL <SEP> NIL <SEP> NIL <SEP> NIL <SEP> NIL <tb> <SEP> 889 <SEP> 23.5 <SEP> NIL <SEP> NIL <SEP> NIL <SEP> NIL <SEP> NIL <SEP> NIL <SEP> NIL <tb> <SEP> 896 <SEP> 33.5 <SEP> NIL <SEP> NIL <SEP> NIL <SEP> NIL <SEP> NIL <SEP> NIL <SEP> NIL <tb> 5* <SEP> 814 <SEP> 32.0 <SEP> NIL <SEP> NIL <SEP> NIL <SEP> NIL <SEP> NIL <SEP> NIL <SEP> NIL <tb> <SEP> 811 <SEP> 25.0 <SEP> NIL <SEP> NIL <SEP> NIL <SEP> NIL <SEP> NIL <SEP> NIL <SEP> NIL <tb> Table 1. Mean Trichostrongylid egg counts in lambs pro and post treatment + Groups 1-3 slaughtered at day 11 post treatment. * Group 5 lambs served for safety evaluation only. TABLE II: Total Worm Counts at Post Mortems of Groups 1-3 EMI15.1 <SEP> N <SEP> o. <SEP> o <SEP> f <SEP> W <SEP> o <SEP> r <SEP> m <SEP> s <SEP> R <SEP> e <SEP> c <SEP> o <SEP> v <SEP> e <SEP> r <SEP> e <SEP> d <tb> <SEP> ABOMASUM <SEP> SMALL <SEP> INTESTINE <SEP> LARGE <SEP> INTESTINE <tb> Groups <SEP> Lamb <SEP> No. <SEP> Early <tb> <SEP> Larval <SEP> Stages <SEP> Adult/Dev.Stages <tb> <SEP> 844 <SEP> 82 <SEP> 19.400 <SEP> 11,900 <SEP> 320 <tb> 1 <SEP> 822 <SEP> 38 <SEP> 1,860 <SEP> 690 <SEP> 10 <tb> <SEP> 866 <SEP> 25 <SEP> 16,700 <SEP> 5,300 <SEP> 80 <tb> <SEP> 812 <SEP> 0 <SEP> 150 <SEP> 380 <SEP> 0 <tb> <SEP> MEAN <SEP> 36 <SEP> 9,528 <SEP> 4,568 <SEP> 103 <tb> <SEP> 870 <SEP> 1 <SEP> 0 <SEP> 0 <SEP> 0 <tb> <SEP> 823 <SEP> 0 <SEP> 0 <SEP> 0 <SEP> 0 <tb> 2 <SEP> 856 <SEP> 0 <SEP> 0 <SEP> 0 <SEP> 0 <tb> <SEP> 880 <SEP> 0 <SEP> 0 <SEP> 0 <SEP> 0 <tb> <SEP> MEAN <SEP> 0.3 <SEP> 0 <SEP> 0 <SEP> 0 <tb> <SEP> % <SEP> reduction <tb> <SEP> compared <SEP> to <SEP> 99.2 <SEP> 100 <SEP> 100 <SEP> 100 <tb> <SEP> controls <tb> <SEP> 851 <SEP> 0 <SEP> 0 <SEP> 130 <SEP> 0 <tb> 3 <SEP> 878 <SEP> 0 <SEP> 0 <SEP> 0 <SEP> 0 <tb> <SEP> 826 <SEP> 0 <SEP> 0 <SEP> 110 <SEP> 0 <tb> <SEP> 2 <SEP> 0 <SEP> 0 <SEP> 0 <SEP> 0 <tb> <SEP> MEAN <SEP> 0 <SEP> 0 <SEP> 60 <SEP> 0 <tb> <SEP> % <SEP> reductin <SEP> 100 <SEP> 100 <SEP> 98.7 <SEP> 100 <tb> <SEP> compared <SEP> to <tb> <SEP> controls <tb> Example 3 Composition per millilitre Fenbenda zole 100.000 mg Fumed silicon dioxide 30.000 mg Sodium methylhydroxybenzoate 1.883 mg Sodium propylhydroxybenzoate 0.205 ing Sodium carboxymethylcellulose 13.720 mg Polyvinylpyrrollidone (PVP) 18.294 mg Sodium Citrate BP 29.837 mg Citric Acid BP 1.649 mg Cobalt Sulphate.7H20 11.925 mg Sodium Selenate.10 H20 9.349 mg Purified Water 880.000 mg Procedure A 2,500 1 mixing vessel was charged with water filtered through a 0.45 ij filter. The following ingredients were added in order, with mixing to disperse them: citric acid, sodium citrate, cobalt sulphate, sodium selenate solution, sodium methylhydroxybenzoate, sodium propylhydroxybenzoate and colloidal silicon dioxide. The following ingredients were added via the in-line homogenisation unit: sodium carboxymethylcellulose, polyvinylpyrrolidone and fenbendazole. Circulation through the in-line homogeniser was continued until a uniform dispersion was achieved. The pH was checked and adjusted to 5.9 with citric acid if necessary. The resulting pink suspensionwas filled into 2 litre containers. Each millilitre of this suspension contains 2.0 mg elemental selenium and 2.5 mg elemental cobalt. This suspension is suitable for cattle. The suggested dose is 7.5 ml per 100 kg body weight. Example 4 Comosition per ml Albendazole 25.000 mg Colloidal silicon dioxide 30.000 mg Sodium methylhydroxybenzoate 1.994 mg Sodium propylhydroxyhenzoate 0.216 mg Sodium carboxymethyl cellulose 14.506 mg Polyvinylpyrrolidone 19.342 mg Sodium citrate 31.590 mg Citric acid 31.350 mg Cobalt sulphate 7H2O 4.500 mg Sodium selenate .1OH2O 3.167 mg Purified water to 1.0 ml Procedure The preparation was made up by a procedure analogous to that described in Example 3. Each ml of suspension contains 0.4 mg of elemental selenium and 0.95 my of elemental cobalt. This suspension is suitable for administration to sheep or goats. The suggested dose is 1 ml per 5 kg body weight. Example 5 Composition per ml Albendazole 100.000 mg Fumed silicon dioxide 30.000 mg Sodium methylhydroxybenzoate 1.883 mg Sodium propylhydroxybenzoate 0.205 mg Sodium carboxymethyl cellulose 13.720 mg Polyvinylpyrrollidone (PVP) 18.294 mg Sodium citrate BP 29.837 mg Citric acid BP 1.649 mg Cobalt sulphate .7H2O 11.925 mg Sodium selenate . 1OE2O 9.349 mg Purified water 880.000 mg Procedure The composition was made up using a procedure analogous to that described in Example 3. Each millilitre of suspension contains 2.0 mg of elemental selenium and 2.5 mg of elemental cobalt. This suspension is suitable for administration to cattle. The suggested dose is 7.5 mg per 100 kg body weight. Example 6 Composition per ml Oxfendazole 22.650 mg Colloidal silicon dioxide 30.000 mg Sodium methylhydroxybenzoate 1.994 mg Sodium propylhydroxybenzoate 0.216 mg Sodium carboxymethyl cellulose 14.506 mg Polyvinylpyrrolidone 19.342 mg Sodium citrate 31.590 mg Citric acid 31.350 mg Cobalt sulphate . 7H2O 4.500 mg Sodium selenate .10 H2O 3.167 ing Purified water to 1.0 ml Procedure The above composition was made up using a procedure analogous to that described in Example 3. Each millilitre of this suspension contains 0.4 mg of elemental selenium and 0.94 mg of elemental cobalt, and is suitable for administration to sheep or goats. The suggested dose is 1 ml per 5 kg body weight Example 7 Composition per millilitre Oxfendazole 90.600 mg Fumed silicon dioxide 30.000 mg Sodium methylhydroxybenzoate 1.883 mg Sodium propylhydroxybenzoate 0.205 mg Sodium carboxymethyl cellulose 13.720 mg Polyvinylpyrrolidone (PVP) 18.294 mg Sodium citrate BP 29.837 mg Citric acid BP 1.649 mg Cobalt sulphate .7H2O 17.887 mg Sodium selenate .10 H2O 14.023 mg Purified water 880.000 ing Procedure The above composition was made up using a procedure analogous to that described in Example 3. Each millilitre of this suspension contains 3.0 mg of elemental selenium and 3.75 mg of elemental cobalt. The suspension is suitable for administration to cattle, and the recommended dose is 5 ml per 100 kg body weight. Example 8 Composition per millilitre Fenbendazole 100.000 ing Fumed silicon dioxide 30.000 mg Sodium methylhydroxybenzoate 1.883 mg Sodium propylhydroxybenzoate 0.205 mg Sodium carboxymethylcellulose 13.720 mg Polyvinylpyrillidone 18.294 mg Sodium citrate 29.837 mg Citric acid 1.649 mg Cobalt sulphate.7H2O 11.925 mg Sodium selenite.5 H2O 6.662 mg Purified water to 100.000 mg Procedure The preparation was made up by a procedure analogous to that described in Example 3. Each millilitre of suspension contains 2.0 mg of elemental selenium and 2.5 mg of elemental cobalt, The suspension is suitable for administration to cattle at a recomrended dose of 7.5 mg per 100 kg body weight. Example 9 (A) The preparation of Example 1 was made up, omitting the selenium and cobalt components, which were replaced by the equivalent weight of purified water. The resulting suspension was filled into 2 litre bottles. Sachets were prepared, each containing 4.5 g of cobalt sulphate.7H2O and 3.17 g of sodium selenate S0H2O. The bottles and sachets were packaged into boxes at the rate of one bottle and two sachet per box,together with instructions (a) to mix the contents of one sachet with one litre of suspension before administration to sheep at a recommended dose of 1 ml per 5 kg body weight, or (b) to mix the contents of each sachet with 1 litre of water, preferably purified water, and to administer the resulting preparation to sheep substantially simultaneously with the anthelmintic suspension. Each being administered at a recommended dose of 1 ml per 5 kg body weight. The selenium/cobalt preparation is preferably administered after the anthelmintic preparation. The contents of the sachet may be made up in a different volume of water, if this is more convenient. The recommended dose should then be calculated pro rata. (B) The preparation of Example 6 was made up analogously to the procedure described in (A) above, i.e. omitting the selenium and cobalt components, which were packaged into sachets as described in (A). The anthelmintic suspension was filled into 5 litre bottles, and bottles and sachets were packaged into boxes at the rate of one bottle and 5 sachets per box. Instructions as set out in (A) were included in each box. (C) The preparation of Example 4 was made up as described in (A) above, but only the selenium component of Example 4 was replaced by purified water. Sachets were made up as described in (A) above, containing 3.17 g of sodium selenate-. 10 H2O. Bottles and sachets were packaged in boxes at the rate of one 2 litre bottle of suspension and two sachets per box. Instructions were included in each box, as specified in (A) above. (D) The preparation of Example 1 was made up as described in (A) above, but only the selenium component of Example 7 was replaced by purified water. Sachets were made up as described in (A), containing 3.17 g of sodium selenate. 1OH2O. Bot-tles and sachets were packaged into boxes at the rate of one 2 litre bottle of suspension and two sachets per box. Instructions were included in each box, as specified in (A) above. Example 10 (A) The preparation of Example 3 was made up with the omission of the selenium and cobalt components, which were replaced by the equivalent weight of purified water. The resulting suspension was filled into 5 litre bottles. Sachets were prepared, each containing 11.9 g of cobalt sulphate 7 H2O and 9.3 g of sodium seienate.10 H2O. The bottles and sachets were packaged into boxes at the rate of one bottle and five sachets per box1 together with instructions to mix the contents of each sachet with either (A) one litre of the anthelmintic suspension or (b) one litre of water, preferably purified water. In the case of (a), the resulting combined preparation is to be administered to cattle at a recommended dose of 7.5 ml per 100 kg body weight. (It is suggested that for convenience, the contents of all five sachets can be admixed with the contents of the 5 litre bottle of suspension). In the case of (b), the instructions point out the requirement for the substantially simultaneous administration of the anthelmintic suspens ion and the selenium/cobalt preparation, and suggest that the seleniun/cobalt preparation is administered immediately after the anthelmintic suspension. The recommended dose for each preparation is 7.5 ml per 100 !g body weight. The instructions also point out that in case (b) the sachets may be made up in a different volume of water, if desired. The recommended dose should then be calculated pro rata. (B) The preparation described in Example 5 was made up as described in (A) above i.e. with the replacement of the selenium and cobalt components by the equivalent weight of purified water. Sachets containing the selenium and cobalt salts were made up as described in (A) above. The anthelmintic suspension, filled into 2 litre bottles, and the sachets, were packaged into boxes at the rate of one bottle and two sachets per box, together with instructions as set out in (A) above. (C) The preparation of Example 7 was made up as described in (A) above but with the replacement of the selenium component only by the equivalent weight of purified water, and filled into 5 litre bottles. Sachets were prepared, each containing 14.02 g of sodium selenate 10H2O- The bottles and sachets were packaged in-to boxes at the rate of one boLtle and five sachets per box, together with instructions. The instructions were as in (A) above except that the recommended dose of the combined preparation or of one selenium sachet made up in one litre of water is 5 ml per 100 kg body weight. (D) The preparation of Example 8 was made up as described in (A) above i.e. with the replacement of the selenium and cobalt components with the equivalent weight of purified water, and filled into 5 litre bottles. Sachets were prepared, each containing 6.662 g of sodium selenite. 5 H2O and 11.925 g of cobalt sulphate.7 H2O. The bottles and sachets were packaged into boxes at the rate of one bottle and five sachets per box, together with instructions as specified in (A) above. (E) The preparation of Example 3 was made up as described in (A) above but with the replacement of only the selenium component and not the con cobalt component by the equivalent weight of purified water, and filled into 5 litre bottles. Cachet were prepared, each containing 9.35 g of sodium selenate. 0H2O. Sachets and bottles were packaged into boxes at the rate of one bottle and five sachets per box. Instructions as set out in (A) above were also included in each box. Example 11 The preparation of Example 3 was tested for anthelmintic efficacy in cattle. There was also used a comparative preparation that did not contain selenium or cobalt. Comparison preparation: Composition per ml. Fenbendazole 100.000 mg Fumed silicon dioxide 30.000 mg Sodium methylhydroxybenzoate 1.883 mg Sodium propylhydroxybenzoate 0.205 mg Sodium carboxymethylcellulose 13.720 mg Polyvinylpyrrollidone (PVP) 18.294 mg Sodium Citrate BP 29.837 mg Citric Acid EP 1.649 mg Purified Water 901.274 m The comparison preparation was made up as described in Example 3. Method A group of 23 heifers was made available for tis study. The cattle had been at grass over the summer and had recently been housed when sampled for allocation to treatment group. Cattle in all three groups were housed in one pen. Faeces samples were collected from all the heifers on two successive days1 and were examined for the presence of worm eggs. On the basis of these worm egg counts the heifers were allocated to one of three treatment groups as shown in Table 1. On the day after second sampling, the cattle were weighed and treated appropriately as indicated in Table 1. Thereafter, faeces samples were collected from the cattle on days 4, 7 and 17 following treatment. Faecal worm egg counts were carried out using the modified McMaster technique. Results The nematode eggs observed in the faeces samples were of the strongyle species and the results of faecal egg counts for all the heifers are shown in Table 2. Both anthelmintic preparations were shown to result in elimination of faecal egg excretion. Conclusions The inclusion of selenium and cobalt does not affect the anthelmintic efficacy of the preparation of Example 3. Table III Faecal Worm Egg Counts Animal klmber prior to treatment Weight Dose Treatment Day Day -2 Day -1 ¯ (kg) (ml) ¯¯¯¯¯ L1282 - 50 350 27 B1175 200 250 380 28 Y1252 400 750 400 30 R 1396 - - 420 32 ) Preparation Q1342 450 550 415 31 ) of Example 3 W1331 500 350 330 26 L1279 300 250 330 26 W1307 550 400 380 28 300 325 Y1267 - - 370 27 L1246 550 450 400 30 W1372 - - 340 26 ) Comparative L1245 250 150 400 30 ) Preparation L1271 450 500 370 37 W1342 450 300 360 27 X 283 233 L1184 50 100 440 L1237 650 400 440 - ) Controls W1340 250 200 300 - ) Untreated W1336 450 600 360 W1281 100 - 380 L1232 - 50 370 W1339 300 200 330 X 257 221 Table IV Animal NhuSber Treatment Faecal Worm Egg Counts following treatments Day 4 Day 7 Day 19 L 1282 B1175 Y1252 R 1395 Preparation of - - Q1342 Example 3 - - W1331 L1279 W1307 Y1267 L1246 W1372 L1245 Comparative - - L1271 Preparation - - W1342 L1184 100 50 * L1237 700 550 550 W1340 Controls 200 200 300 W1336 Untreated 450 450 400 W1281 50 - L1232 - - * W1339 300 350 200 X 257 228 290 *Not sampled Example 12 The safety of the composition of Example 3 on oral administration to cattle was investigated: A total of 16 cattle were treated orally with the preparation of Example 3. Eight Friesian heifers weighing between 330 and 420 kg were treated with the preparation at the recommended dose of 7.5 ml per 100 kg body weight. Four Friesian cattle weighing between 930 and 990 kg were treated orally with the preparation at five times the recommended dose i.e. 37.5 ml per 100 kg. A further four Friesan cattle weighing between 870 and 1030 kg were treated orally with the preparation at ten times the recommended dose i.e. 75 mg per 100 kg. Table 1 shows the computed and actual dose administered relative to live weight. The cattle were observed daily for a period of 21 days following treatment for any signs at adverse effect. Results and Conclusions No signs of adverse toxicity were observed in cattle treated with up to ten times the recommended dose of the formulation. Oral administration of up to 20 mg/kg of selenium and 25 mg/kg of cobalt in combination with 75 mg/kg of fenbendazole was carried out in cattle with no adverse effect observed. Table V Volumes of preparation of Example 3 administered in safety tests. EMI28.1 <tb> Animal <SEP> Number <SEP> Weight <SEP> (kg) <SEP> Calculated <SEP> Dose <SEP> Dose <SEP> Adminis <tb> <SEP> (ml) <SEP> tered <SEP> (ml) <tb> L1282 <SEP> 350 <SEP> 26.2 <SEP> 27 <tb> B11 <SEP> 380 <SEP> 28.5 <SEP> 28 <tb> Y1252 <SEP> 400 <SEP> 30.0 <SEP> 30 <tb> R1396 <SEP> 420 <SEP> 31.5 <SEP> 32 <tb> Q1342 <SEP> 415 <SEP> 31.1 <SEP> 31 <tb> W1331 <SEP> 330 <SEP> 24.8 <SEP> 26 <tb> L1279 <SEP> 330 <SEP> 24.8 <SEP> 26 <tb> W1307 <SEP> 380 <SEP> 28.5 <SEP> 28 <tb> 2 <SEP> 990 <SEP> 372 <SEP> 380 <tb> 7 <SEP> 950 <SEP> 357 <SEP> 360 <tb> 71 <SEP> 930 <SEP> 349 <SEP> 360 <tb> 10 <SEP> 930 <SEP> 349 <SEP> 360 <tb> 65 <SEP> 870 <SEP> 652 <SEP> 660 <tb> 17 <SEP> 1030 <SEP> 774 <SEP> 780 <tb> 87 <SEP> 960 <SEP> 721 <SEP> 720 <tb> 90 <SEP> 925 <SEP> 695 <SEP> 700 <tb>

Claims: Patent Claims: 1. A veterinary preparation comprising an effective amount of an anthelmintic agent and a non-toxic amount of selenium salt effective to prevent selenium deficiency in an animal. 2. The veterinary preparation of claim 1, wherein said anthelmintic agent and selenium salt are in admixture. 3. The veterinary preparation of claim 1, additionally comprising a solid or liquid carrier suitable for use in a veterinary preparation. 4. The veterinary preparation of claim 1, additionally comprising a non-toxic amount of cobalt salt effective to prevent cobalt deficiency in an animal. 5. The veterinary preparation of claim 4, wherein said anthelmintic agent is fenbendazole. 6. The veterinary preparation of claim 5, wherein said selenium salt is a soluble selenium salt selected from the group consisting of sodium selenate, potassium selenate, ammonium selenate, sodium selenite, potassium selenite, ammonium selenite, sodium selenite, potassium selenide and ammonium selenide. 7. The veterinary preparation of claim 5, wherein said cobalt salt is a soluble cobalt salt selected from the group consisting of cobalt chloride, cobalt carbonate, cobalt nitrate and cobalt sulphate. 8. The veterinary preparation of claim 4, wherein said anthelmintic agent is oxfendazole. 9. The veterinary preparation of claim 4, wherein said anthelmintic agent is albendazole. 1 0. The veterinary preparation of claim 4, wherein said anthelmintic agent is selected from the group consisting of thiabendazole, parbendazole, cambendazole, oxi bendazole, mebendazole and fluobendazole. 11. The veterinary preparation of claim 4, wherein said selenium salt is a soluble selenium salt selected from the group consisting of sodium selenate, potassium selenate, ammonium selenate, sodium selenite, potassium selenite, ammonium selenite, sodium selenide, potassium selenide and ammonium selenide. 12. The veterinary preparation of claim 4, wherein said cobalt salt is a sol tib] e. cobalt salt selected from tie group consisting of cobalt chloride, cobalt carbonate, cobalt nitrate and cobalt sulphate. 13. The veterinary preparation of claim 1, wherein said anthelmintic agent is fenbendazole. 14. The veterinary preparation of claim 1, wherein said anthelmintic agent is oxfendazole. 15. The veterinary preparation of claim 1, wherein said anthelmintic agent is albendazole. 16. The veterinary preparation of claim 1, wherein said antlielmintic agent is selected from the group consisting of thiabendazole, parhendazole, cambend azole, oxibend zole mebendazole and fluobendazole. 1 7. The veterinary preparation of claim 1, wherein said selenium salt is a soluble selenium salt selected from the group consisting of sodium selenate, potassium selenate, ammonium selenate, sodium selenite, potassium selenite, ammonium selenite, sodium selenide, potassium selenide and ammonium selenide. 18. A veterinary preparation for sheep and goats comprising an effective amount of an anthelmintic agent and a non-toxic amount of selenium salt effective to prevent selenium deficiency in sheep and goats, wherein said anthelmintic agent is present in an amount of from 100 to 400 parts by weight; and wherein said selenium salt is present in an amount, calculated as elemental selenium, of 1 to 20 parts by weight. 19. The preparation of claim 18 additionally comprising a non-toxic amount of cobalt salt effective to prevent cobalt deficiency in sheep and goats, wherein said cobalt salt is present in an amount, calculated as elemental cobalt, of 1 to 50 parts by weight. 20. A veterinary preparation for cattle comprising an effective amount of an anthelmintic agent and a non toxic amount of selenium salt effective to prevent selenium deficiency in cattle, wherein said anthel mintic agent is present in an amount of from 150 to 1500 parts by weight; and wherein sad selenium salt is present in an amount, calculated as elemental selenium, of 0.75 to 18.75 parts by weight. 21. The preparation of claim 20, additionally comprising a non-toxic amount of cobalt salt effective to prevent cobalt deficiency in cattle, wherein said cobalt salt is present in an amount, calculated as elemental cobalt, of 0.75 to 37.5 parts by weight. 22. A method of treating a ruminant comprising the step of administering to said ruminant a non-toxic amount of the preparation of claim 1 effective both to produce an anthelmintic result in said ruminant and to prevent selenium deficiency in said ruminant. 23. The method of claim 22, wherein said ruminant is selected from the group consisting of sheep and goats and wherein said anthelmintic agent is administered in an amount of from 100 to 400 mg/50 kg body weight of said ruminant and wherein said selenium salt is administered in an amount, calculated as elemental selenium, of 1 to 20 mg/50 kg body weight of said ruminant. 24. The method of claim 22, wherein said ruminant is cattle and wherein said anthelmintic agent is administered to said cattle in an amount of from 150 to 1500 mg/100 kg body weight of said cattle; and said selenium salt is administered to said cattle in an amount, calculated as elemental selenium, of 0.75 to 18.75 mg/100 kg body weight of said cattle. 25. A veterinary pack comprising an anthelmintic agent and a selenium salt, said anthelmintic agent and said selenium salt being separately packaged and said anthelmintic agent and said selenium salt being combinable to form a veterinary preparation comprising an effective amount of an anthelmintic agent and a non-toxic amount of selenium salt effective to prevent selenium deficiency in an animal. 26. The veterinary pack of claim 25 additionally comprising a cobalt salt, said cobalt salt being separately packaged from said selenium salt and said anthelmintic agent and said anthelmintic agent, cobalt salt and selenium salt being combinable to form a veterinary preparation comprising an effective amount of an anthelmintic agent, a non-toxic amount of selenium salt effective to prevent selenium deficiency in an animal and a non-toxic amount of cobalt salt effective to prevent cobalt deficiency in an animal. 27. A veterinary pack for sheep and goats comprising an anthelmintic agent and a selenium salt, said anthel mintic agent and said selenium salt being separately packaged and said anthelmintic agent and said selenium salt being combinable to form a veterinary preparation for sheep and goats comprising an effective amount of an anthelmintic agent and a non-toxic amount of selenium salt effective to prevent selenium deficiency in an animal; wherein said anthelmintic agent is present in said pacl; in an amount of from 100 to 400 parts by weight; and wherein said selenium salt is present in said pack in an amount, calculated as elemental selenium, of 1 to 20 parts by weight. 28. The veterinary pack of claim 27 additionally comprising a cobalt salt, said cobalt salt being separately packaged from said selenium salt and said anthelmintic agent and said anthelmintic agent, cobalt salt and selenium salt being combinable to form a veterinary preparation for sheep and goats comprising an effective amount of an anthelmintic agent, a non-toxic amount of selenium salt effective to prevent selenium deficiency in an animal and a non-toxic amount of cobalt salt effective to prevent cobalt deficiency in an animal; wherein said cobalt salt is present in said pack in an amount, calculated as elemental cobalt, of r to 50 parts by weight. 29. A veterinary pack for cattle comprising an anthelmintic agent and a selenium salt, said anthelmintic agent and said selenium salt being separately packaged and said anthelmintic agent and said selenium salt being combinable to form a veterinary preparation for cattle comprising an effective amount of an anthelmintic agent and a non-toxic amount of selenium salt effective to prevent selenium deficiency in an animal; wherein said anthelmintic agent is present in said pack in an amount of from 150 to 1500 parts by weight; and wherein said selenium salt is present in said pack in an amount, calculated as elemental selenium, of 0.75 to 18.75 parts by weight. 30. The veterinary pack of claim 29 ajditionally comprising a cobalt salt, said cobalt salt being separately packaged from said seven: tun salt. and said anthelmintic agent and said anthelmintic agent5 cobalt salt and selenium salt being combinable to form a veterinary preparation for cattle comprising an effective amount of an anthelmintic agent, a non-toxic amount of selenium salt effective to prevent selenium deficiency in an animal and a non-toxic amount of cobalt salt effective to prevent cobalt deficiency in an animal; wherein said cobalt salt is present in said pack in an amount, calculated as elemental cobalt, of 0.75 to 37.5 by weight.

Inventors: GANLEY, JOHN ANTHONY, MCBEATH, DAVID GEORGE, MCTAGGART, CELIA MARGARET, WELLS, PETER WILLIAM
Applicants: HOECHST UK LIMITED
Application year: 1983
Main classification: A61K33
IPC codes: A61K33