ANIMAL FEED FUR KALBER

The invention concerns the outer/breed of calves and in particular the creation of conditions, in order to decrease their AnfflHgkeit in relation to diseases of the intestine tract and in particular opposite such getting sick eyes, which consequence after the first life days of a tightened infection are. Such an intestine illness, which outer-rode with calves, is the result of an infection by pathogene Sf e von Bacterium Escherichia coli, and in particular such trunks, which possess the Serotypenbezeichnungen 08, 09, 015, 078, 0114, 0137 and 0139 (internationally Serotypc Classification). In brit. Patent specification No. 760.691 was suggested the production the Wachsünn of promoting animal feed additive for chickens, which from a culture of a living E-CO] to i-organism (a serelogischen kind bestirnten more near), which had been isolated from blind intestine contents from Hü3nern, and one eru, the Wachsbnn promoting not bakterieilen substance, preferably an antibiotic, like a penicillin, Oxytetracyclin, Aureomyein do not hrenden or Bacitracin, exists. In accordance with revealing the Patentschriß mentioned the living E-coli-Organiemus and the Antibioßkum affect each other mutually, how a large growth is obtained as by the antibiotic A! flax, against what there is no reciprocal effect between the dead organism and the antibiotic. If the environment or the Fußer is changed by calves, for example in the case of their transport, which causes admission into the stable or when taking away the dam the associated stress situations, da8 the calves in relation to the increase of pathogenen organisms is less widersiandsfähig, and it exists to cause a large Wahrscheinlinhkeit, Trade Union of German Employees any bacteria of E. coli, which were taken up with the food, a heavy failure (diarrhea). Task of the invention is it to supply here a means. Now, the intestine from calves was surprisingly found to Trade Union of German Employees to the production of antibodies against pathogene Organtsmen of the trunks of E, specified above. coli lively will can, by the Endotoxine of the organisms, practically freely of the living organisms the calf ora! , i.e. differently of the Einfü] lrungsweise are given into the Blutkreislauf as for example by injection. Those would invention-in accordance with-eat task by an oral NIitüel to the raising of calf solved, which is characterised by it, Trade Union of German Employees it the Endotoxine one pathogenea, on the intestine tract of calves of infectious working bacterium contains. With designation “Endotexine” for in intestine of calves to be saved materials is not said that excluded in these materials the presence from Endotoxinen or the presence of the Zellmaserials, in which the Endotoxine are enclosed in the living Baeterium its sol!. By this it is to be only understood that Endotoxine have the Hauptbedeuümg with the achievement of the gewünschtenimmunologfschen effect, while and would backfind Exotoxine the cells this desired immunological effect does not exhibit. However k nn it if necessary more comfortably its to have either Exotoxine or cell arrears or both with the Endotoxinen together available to save first in order the work of their isolating and danu also, in order to use the antigens the effects, which can possess it. Feeding the calf with the Endotexinen of only one specified the before spurs from E. eoli has to that extent a certain favorable effect to arrange as the ability of this trunk damage is decreased. However it is preferred to applizieren the Endotexine of all voice of E. coli. Too aplr lozierenden Endotoxine can by breeding each of these trunks, whereby eime sample of a j of eden trunk of the Cen ral Veterinary Laboratory, Ministry OF Agriculü RH and Fisheries, new Haw, Weybridge, Surrey, Great Britain or of that is national Colleetton OF type Cultures erhälülieh, and after which progressing the growth of the 1Vlikroorganismus and thus the production from Endotoxlnen to a suitable extent by killing the increasing microorganisms and release of the Endetexine will receive. Latter kanu by Abkochen or by treatment in autoclaves to be reached. The entire, sterilized cultures, about which each Exotexine and the cell arrears contain like also the EN dotoxine, can be together-given and appliziert then. Alternatively can however also instead of the sterilisation of the entire Kulhu: EN the bacteria separated, usually by centrifugation, and in a small volume of an aqueous medium, e.g. in water or a salt solution, for killing the bacteria and for the release of the Endotoxinebehandeltwerden. The simplest and most economical way for killing the separated bacteria is heating up. If heating up persists sufficiently for a long time, e.g. 1 h with 100°C, 20 min bei125°C, becomes a rough part of the Endotoxins from its connection with the CPU! lwänden the killed bacteria solved and into the medium, in which heating up is accomplished, set free. The remaining, bound Endotoxine can if necessary by treatment with an enzyme such as Trypsin in solution bring will, as this is descriptive e.g. in the Netherlands patent specification No. 7107538. Of like the received Endotexinen besehrieben above animal feeds for calves in firmer or in liquid form can be made, by mixing the Endotoxine with or several of the vital components of the fodder of the calves, i.e. protein, fat, coal hydrate, vitamins and mineral elements, e.g. calcium and phosphorus. In this way the Enäotoxine for the application can with 1V [agermilchpulver, grain or mineral elements to be mixed, or they can be added to a complete FuLtermiLtel, i.e. a animal feed, which contains all vital components of the food such as protein, February etc. The invention is with the increase of the ruggedness in relation to infection of calves of special Bedeuümg, if these completely particularly onen from the Mußertter taken away and as consequence hievon stress Siü t are suspended. By beginning input Endotoxinmaterials in freon age” be suitable-proves with 4 to 10 days, and preferably to last feeding time with dam or briefly thereafter and by sport guidance during appropriate time-save, vorteilhafterv¢eise up to age von4 to 6 weeks, l nn intestine calves to production suitable antibodies lively become, so that at the time, at which the calf is exposed to a strong Infektiunsgefahr, a sufficiently high circulation of antibodies is present in the intestine, in order to become finished either with the increase of any, with the food taken up, pathogener bacteria or to lower at least the weight of any Erkranlmngszustandes, which develops nevertheless. The Endotoxinmaterial can be entered for this purpose into a so-called “milk substitute”, i.e. into the food, which is particularly zusaramengesteltt, in order to carry the Ernährungsbedürfmissen of calves calculation, if they are taken away from the dam. It can be likewise entered into the first, dry food, which is given in connection with milk or a milk substitute when curing. It is favourable, the Endotoxine in form of a dry, firm Vormischung with the odermchreren food components a protein, February, to make coal hydrate or mineral Elémenten available whereby the consumer mixes this Vormischung with the remaining components of the fodder. If necessary Endotexine of the Sahnonellenst can do mmeS.dublinundS typhimurium zuspannen with those of the trunks of E. coli containing foot-determine to be manufactured. The minimum dose rates lie within the range of 1 to 10 units of the Endotexine of each bacteria type per calf per day. It was however found that calves, to which the 10O0fache quantity of this value was given did not show symptoms. A preferredwise maximum for an input in einFuttermißel for calves amounts to 1000 units of the Endotexine of each Serotyp per kg of the Futtereißels. The units are measured as follows: Production and isolation of Endotoxin for the standard regulations. Endotoxine can be mmen of the Salraonellens and be made the 7 Serotypen by E. coli according to the following method, which generally with the method of westchamfer, Ruderitz and Bister, Z. nature-vigorously. 7 [195 P. 148 agrees. A freezingdried culture iVfikroergaräsmus in peptone water one pours and one inkubtert 6 h bei37°C. Examined after a suitable Wachsünn the culture for their purity on washed shank bußgarplatte and then used for the inoculation of diagonal feeding agar (Oxoid) in Roux bottles. Oxoidnährbrühe No. 2 is a well-known culture medium. To its production 10 g beef excerpt, 10 g peptone and 5 g sodium chloride are solved in 1 1 distilled water and the received solution (pH 7, 5) in autoclaves with 1 RK a positive pressure (121°C) 15 min long heated up. The culture is let grow over night with 37°C, and the bacteria are collected in sterile, distilled water. The received bacteria suspension is given aseptisch in 30 ml universal bottles and centrifuged with 4000 Umdr/min during 10 min. The supernatant liquid will become removed and the Bakterienk'ügelchen staying in 4 ml sterile, distilled water again suspension and then freezingdried. For the isolation of the Endotoxine 0.5 g of the freezingdried material in 5 ml 0, are suspended 15 M NaC1 and 10 ml 90%iges aqueous phenol are in addition-given as lysierendes means. The mixture is warmed up to 30 min to 68°C under continuing grooves, centrifuged afterwards it with 4000 Umdr/min for consolidating the Zellrfickstände. The aqueous phase will become removed and auf4OC abgeknallt, hiezu slowly 10 VOL from ice-cooled Ätt nol in addition-given. In such a way formed precipitation will become in water again dissolved and the groove your acids from the solution by addition removed from 2 VOL Äihanol precipitated and. From the Resflösungwerden the Endotexine by addition precipitated by further 8 VOL ethanol. They are separated by centrifugation, washed with ice-cooled ethanol and again dissolved in 0, 15 1Vr NaC1. This solution of the Endotoxine is called in the following reagent 1. Investigation of Endotoxinen • a) production of anti-serum specific, hyperiramune serums (anti-serums) New Zealanders rabbits become against washed, by heat killed in white, (10 0°C, 2, 5 h) organisms of everyone the Serotypen of E. coliund Salmonella produced. Suspensions of the organisms killed by heat are manufactured in 0,15 M NaC1 (about 3 x10 9 organisms/valley), and injected intravenously. The Tmmunisierungsprogr mm begins with a InJel onsvolumen from 0,1 ml and on each fourth day under doubling of the volumes to 1, 6 ml is continued. 10 days after conclusion of the program the blood is taken by the rabbits and this received blood is centrifuged. The upper layer of the hyperimmune serum is collected. This anti-serum is called reagent 2. b) Measurement of the antibodies (anti- Endotexine) activity in the anti-serum erythrocyte of the sheep by treatment of a 5% suspension of washed, packed cells with a same volume of the Endof xinlösung reagent 1 with 37°C during 30 min sensibilisiert0 the sensitized toes are separated by centrifugation, washed freely from surplus Endotexinen and suspended again in 0, 15 M NaC1 under education 2, of a 5% suspension from Schaferythrocyten sensitized with Endotexin. This suspension is called reagent 3. The reagent 2, i.e. the anti-serum, is diluted in sequence with 0,15 M NaCI, around a set of L8sungen of same volume (1 mol) with one anti-body concentration of 1/5, 1/10, 1/20, 1/40… to receive 1 (5 x 2n-l) that from reagent 2, and too 1/5 Voi the reagent of the 3 are given to each of these solutions. In the stronger Antiserumtösungen a Hämagglutination arises, however not in the weaker solutions, and as terminator point of titration accomplished with 4oC the solution is regarded, in which the Hämagglutinatfon straight still outer-rode. With a typical function the terminator point preserves with the zwöli Len solution, i.e. with the solution, which possesses an anti-body concentration of 1 (5 x211) that from reagent 2. Therefore an anti-serum titer can be attributed to the reagent 2 by 5 ×2t t (=10 240). c) Measurement of the Endotexinkonzentratien in solution of unknown quantities concentration the solution which can be examined (Y) with 0, i5 M NaC1 is diluted in sequence, in order to receive a set of solutions of same volume (3 VOL) with the Enäotoxinkonzentratio EN 1/3, 1/ö, 1/12, to 1/24 etc. of those the solution Y. Too each of these solutions becomes 1 VOL the reagent the 2 (Antisertun), which was diluted in such a way that it a titer of 20 (S. unte before, “b) possesses, and then after some IVIinutenl VOL the reagent of the 3 (Schaferyfhrocyten sensibillsierte) in addition-given, whereby in the long run Endotexinkonzentraßonen will receive i (5 ×2n-l) of that ven solution Y from 1/5, I/I0, 1/20…. The Hämagglutination is inhibiert in the stronger Endotexinlösungen, arises however in the schwäeheren, and the terminator point of titration is accepted with the solution, in which a Hämagglutination straight is still prevented. With a typical function this took place with the sixth solution with a Endotoxinkonzentration = 1 (5 x 2) that the solution Y. from this it results that the solution Y possesses a titer of 5 x2 (= 160), the 160 units Endotexin/ml is äqnivalent). Example: A. Production of raw Endoto umaterial the following function was durchgefifllrt separately with everyone the Salmonellenst mme and everyone the Serotypen from E. coli, which were called before. (i) the Batderium by a laid down Vorratsladtur on washed blood agar plates in case of of E. madly and on feeding agar plates in case of of Salmonella was applied and 24 h with 37°C inkubierL the plates were then examined in an appropriate way on purity of the trunk. (IL) Colonies of the bacterium became of the plates in 50 ml Oxoid Nährbrühe No. 2 (catalog No. cm 67) (composition see above) übergüllrt. The Brühe and/or Nährglüssigkeit one held for 24 h bei37°C. (iii) the entire under (IL) received Kulttu: for inoculating 1, 5 ml Oxoid Nährbrü e No. 2 were used, and the nutrient fluid was inlmbiert under vibrating 24 h bei37°C. Each in this way erhaitene EN tur contained 10 9 _ of 1010 life NSF l ige bacteria/rel. Samples of the cultures of E. coä resulted in, if they had been treated 2 h beß 125oc with steam and as were before descriptive subjected to the research method, a titer of 2560, which corresponds to 2560 units Endotexin per ml the culture treated with steam. Samples of the cultures of S. Dublin and S. typhimurium were bchandelt equally as the cultural gene by E. coli with steam and examined and resulted in a titer of 1280. Everyone of the new cultures, i.e. 7 kinds of E. eoli and one the Salmonellen Serotypen, were zensuriert in each case for the separation of the bacteria, which were then suspended partly again in the separated, supernatant utterflüssigkeit, around a suspension with a 30Bchen concentration of those the culture before the Zenü: ifugieren to receive. Each suspension was separated 2 h in a Autok! aven with 1250C for killing the bacteria treated with steam, then the new suspensions treated with steam were entered zusam. B. Production of the oral means for calves, the together-given material treated received under A with steam (i thread - part) was mixed with 19 parts of whey powders, and the developed mixture was entered in a usual Mllchersatzsteff of the following composition: Thread - % skimmed milk powder with added fat whey powder glucose gefällter lime Dikalziumphosphat trace minerals and vitamins on carriers in the received product the concentration of the Endotoxine amounted to: 77 17.1 0.3,0.4,0.2 Endotoxine of each Serotyp of E. coli Endotexine of each Serotyp of Salmonella 100 Einheiten/kg of the animal feed Einheiten/kg of the Futtennittels Nr.328280