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Zusammenfassung Im aktiven Mauseschutzversuch wurde einmal vergleichend die Immuno-genitat von formalin- und warmeinaktivierten P. multocida-Keimen und zum anderen, am Beispiel des “warmeinaktivierten” Impfstoffes, die Immunogenitat verschiedener P. multocida-Stamme, sowie die Wirksamkeit einer oralen, intranasalen und subkutanen Impfung untersucht. Die Wirksamkeit der P. multocida-Impfstoffe war unabhangig vom In-aktivierungsverfahren. Deutliche Wirksamkeitsunterschiede ergaben sich nur bei Verwendung unterschiedlicher P. multocida-Stamme fur die Impfstoff-herstellung. Der Impfstoff aus “warmeinaktivierten” Keimen war sowohl bei sub-kutaner als auch bei oraler und intranasaler Impfung wirksam. Die erzielten Schutzwerte lagen bei allen 3 Applikationsarten deutlich uber dem von Ose und Muenster geforderten Mindestschutzwert. Die dafur notwendigen Impfdosen betrugen 1,25 · 1011fur die zehnmalige orale, 2,5 · 1010fur die zwei-malige subkutane und 3 · 109inaktivierte Keime fur die einmalige intranasale Impfung. Summary Efficacy and harmlessness of P. multocida-vaccines based on inactivated bacteria after subcutaneous, oral and intranasal application in mice In comparative investigations using the mouse protection test 1. immuno-genicity of formalin- and heat-inactivated P. multocida, 2. immunogenicity of different P. multocida-strains and 3. efficacy of oral, intranasal and subcutaneous vaccination with heat-inactivated bacteria were studied. The efficacy of P. multocida-vaccines was independent of the inactivation procedure. Distinct differences in efficacy were only observed after the use of different P. multocida-strains for vaccine production. The “heat-inactivated” vaccine was efficient both after subcutaneous and after oral and intranasal vaccination. With all application methods the protection rates obtained were higher than the required minimal protection rate (Ose and Muenster). The necessary vaccination dosages amounted to 1.25 · 1011for ten times oral, 2.5 · 1010for twice subcutaneous and 3 · 109inactivated bacteria for single intranasal vaccination. Resume Action et inocuite de vaccins Pasteurella multocida a partir de germes inactives apres une application sous-cutanee, orale et intranasale chez la souris On a compare l'immunogenicite chez des souris de germes P. multocida inactives a la formaline et a la chaleur en prenant un vaccin «inactive a la chaleur», l'action immunogene de differentes souches de P. multocida et l'effet d'une vaccination orale, intranasale et sous-cutanee. L'effet des vaccins a P. multocida fut independant du procede d'inactivation. De nettes differences ont ete constatees lors de l'emploi de differentes souches de P. multocida pour l'elaboration du vaccin. Le vaccin «inactive a la chaleur» fut aussi efficace par voie sous-cutanee qu'orale et intranasale. Les valeurs de protection obtenues avec les 3 modes d'applications se situerent nettement au-dessus des valeurs de protection recommandees par Ose et Muenster. Les doses necessaires ont ete de 1,25 · 1011pour la vaccination orale en dix fois, de 2,5 · 1010pour l'application sous-cutanee en deux fois et de 3 · 109germes inactives par voie intranasale en une fois. Resumen Eficacia e innocuidad de vacunas Pasteurella multocida a partir de germenes inactivados tras aplicacion subcutenea, oral e intranasal en el raton En el ensayo de proteccion activa de ratones se estudio por un lado comparativamente la inmunogenia de los germenes P. multocida inactivados con formol o calor, y por otro, en el ejemplo de la vacuna «termoinactivada», la inmunogenia de varias estirpes P. multocida, asi como la eficacia de la vacunacion oral, intranasal y subcutanea. La eficacia de las vacunas P. multocida no dependia del procedimiento de inactivacion. Solo se infirieron diferencias marcadas en la eficacia utilizando diversas estirpes P. multocida para la elaboracion de las vacunas. La vacuna a base de germenes «termoinactivados» era eficaz tanto tras la aplicacion oral como la intranasal. Los valores protectores conseguidos se hallaban en todas las 3 modalidades de aplicacion claramente por encima del valor protector minimo exigido por Ose y Muenster. Las dosis vacunales precisas para ello fueron del orden de 1,25 · 1011germenes inactivados para la vacunacion oral en diez ocasiones, 2,5 · 1010para la vacunacion subcutanea en dos ocasiones y 3 · 109para la vacunacion intranasal hecha una vez sola.

Summary Eighteen Caesarian-derived, colostrum-deprived calves were orally infected with either bovine rotavirus, enterotoxigenic Escherichia coli (ETEC) or with a combination of both enteropathogens within the first 24 h of life. Animals which received ETEC developed severe diarrhea when 1010organisms were given, but remained healthy after inoculation of 107organisms. Animals infected with four different field strains of bovine rotavirus developed mild, transient diarrhea after an average incubation time of 29 h. They usually recovered within a few hours after onset of the symptoms. A synergistic effect was observed after simultaneous application of rotavirus and 107ETEC, resulting in severe diarrhea after an average incubation time of 22 h. These calves had significantly more rotavirus in the intestinal samples than to samples from monoinfected calves. Those calves, which were infected with 107ETEC followed by rotavirus 15 to 36 hours later, had significantly less rotavirus in the intestinal samples and shed significantly less rotavirus in their feces than to monoinfected calves. The number of ETEC organisms demonstrated in the small intestine was more dependent on the time of sampling than on the mode of infection. Zusammenfassung Synergismus nach experimenteller Infektion neugeborener Kalber mit bovinem Rotavirus und enterotoxischen Escherichia coli (ETEC) Achtzehn Kalber wurden innerhalb der ersten 24 Lebensstunden oral entweder mit bovinem Rotavirus, enterotoxinbildenden E. coli (ETEC) oder mit einer Kombination aus beiden Erregern infiziert. Tiere, die 1010ETEC erhielten, entwickelten eine schwere Durchfallerkrankung, wahrend 107ETEC keine Durchfallsymptome mehr auslosten. Tiere, die mit vier verschiedenen Rotavirus-Feldstammen infiziert wurden, zeigten nach einer durchschnittlichen Inkubationszeit von 29 Stunden eine milde, kurzzeitige Diarrhoe. Sie erholten sich wenige Stunden nach Sistieren des Durchfalls. Nach gleichzeitiger Infektion von Rotavirus mit 107ETEC wurde eine synergistische Interaktion beider Erreger beobachtet. Diese Kalber entwickelten eine schwere Durchfallerkrankung nach einer durchschnittlichen Inkubationszeit von 22 Stunden. In den Darmabschnitten dieser Tiere wurden signifikant hohere Rotavirusinfektionsitatstiter gefunden als bei Tieren, die mit Rotavirus allein infiziert wurden. Dagegen war die Virusausscheidung im Kot und in den Darmabschnitten signifikant verringert bei Tieren, denen zuerst 107ETEC und 15 bis 36 Stunden spater Rotavirus verabreicht wurde. Die Zahl der im Dunndarm festgestellten ETEC-Keime war in allen Gruppen starker vom Zeitpunkt der Probengewinnung nach Infektion als von der Art der Infektion abhangig. Resume Synergisme apres une infection experimentale de veaux nouveaux-nes avec un Rotavirus bovin et Escherichia coli enterotoxique (ETEC) Des infections mixtes experimentales ont ete faites sur des veaux nouveaux-nes, prives de colostrum avec un Rotavirus bovin et E. coli enterotoxique (ETEC). Dix-huit veaux ont ete infectes oralement dans les premieres 24 heures de vie avec un Rotavirus bovin ou E. coli enterotoxique (ETEC) ou avec une combinaison des deux agents. Les animaux ayant recu 1010ETEC developperent une grave diarrhee alors que 107ETEC ne provoquerent aucun symptome de diarrhee. Les animaux qui furent infectes avec quatre souches differentes de Rotavirus du terrain presenterent une legere et breve diarrhee apres un temps d'incubation de 29 heures. Ils ont ete gueris quelques heures apres la cessation de la diarrhee. Une interaction synergique des deux agents a ete observee apres une infection simultanee de Rotavirus avec 107ETEC. Ces veaux developerent une grave diarrhee apres une periode d'incubation moyenne de 22 heures. On a trouve des titres infectieux de Rotavirus significativement plus eleves dans les portions d'intestin de ces animaux que chez les veaux infectes seulement avec du Rotavirus. L'excretion virale dans les matieres fecales et dans les portions intestinales fut par contre significativement diminuee chez les animaux qui avaient recu d'abord 107ETEC et Rotavirus 15 a 36 heures plus tard. Le nombre des germes ETEC etablis dans l'intestin grele avec tous les groupes a dependu plus du moment des prelevements apres l'infection que du genre de l'infection. Resumen Sinergismo tras infeccion experimental de terneros recien nacidos con rotavirus bovino y Escherichia coli enterotoxica (ETEC) Dieciocho terneros que no recibieron calostro fueron infectados dentro de las 24 horas primeras de vida por via oral bien con un rotavirus bovino, E. coli enterotoxinogena (ETEC) o bien con una combinacion de ambos agentes etiologicos. Los animales que recibieron 1010ETEC desarrollaron una enteritis grave, mientras que 107ETEC no desencadenaron sintomas diarreicos. Los animales que fueron infectados con cuatro estirpes campales diferentes de rotavirus mostraron tras un tiempo medio de incubacion de 29 horas una diarrea benigna de duracion breve. Los mismos se recuperaron pocas horas despues del cese de la descomposicion. Tras la infeccion simultanea de rotavirus con 107ETEC se observo una interaccion sinergica de ambos agentes etiologicos. Estos terneros desarrollaron una enteritis grave tras un periodo medio de incubacion de 22 horas. En los tramos entericos de estos animales se hallaron titulos de infecciosidad significantemente mas elevados que en los animales que habian sido infectados solo con rotavirus. Sin embargo, la eliminacion de virus con las heces y en los tramos intestinales se hallaba reducida de forma significativa en los animales que recibieron primero 107ETEC y 15 hasta 36 horas despues rotavirus. La cuantia de los germenes ETEC comprobada en el intestino delgado dependia en todos los grupos mas del momento en que se recogia la muestra tras la infeccion que del tipo de infeccion.

Zusammenfassung Vergleichende Untersuchungen mit Hilfe des CA-Testes an Frenkel- und BHK-allergischen Rindern bestatigen, das fur die nach einer mehrmaligen MKS-Schutzimpfung auftretende Allergie vom Spattyp sowohl das MKS-Virus allein als auch in Kombination mit allergenen BHK-Zellbestandteilen verantwortlich sein kann. Das MKS-Virus allein wirkt nur schwach allergen. Enthalten MKS-Vakzinen allergene BHK-Zellanteile, so bringen sie das Virus-Allergen im Sinne eines Adjuvans-Effektes starker in Reaktion. Durch Aufspaltung des Virus gewonnene Virus-RNS war nicht allergen. Das isolierte Virus-Protein dagegen besas eine stark allergene Wirkung. Modellversuche an sensibilisierten Meerschweinchen bestatigten diese Beobachtung. Meerschweinchen konnten sowohl mit komplettem, gereinigtem MKS-Virus, mit abgespaltenem MKS-Virus-Protein als auch mit BHK-Zellsubstanzen sensibilisiert werden. Die Allergie lies sich durch Makrophagen ubertragen. Die MKS-Allergie war in der Regel schwacher ausgebildet als die BHK-Allergie. Eine Ausnahme bildeten die Meerschweinchen, welche mit hochgereinigtem und konzentriertem Ultrazentrifugen-Virus-Pellet sensibilisiert worden waren. Hier traten im CA-Test praktisch nur noch virusspezifische Reaktionen auf. Mit reiner Virus-RNS konnten die Meerschweinchen nicht sensibilisiert werden. Das aus hochgereinigtem MKS-Virus abgespaltene Protein sensibilisierte die Meerschweinchen in starkerem Mase als das komplette Virus. Im Kreuz-CA-Versuch an Meerschweinchen, die gegen die Virustypen O1, A5 oder C1 sensibilisiert worden waren, reagierten A5 und C1 starker miteinander als mit O1. Dieser Typ besas dagegen die starkste sensibilisierende Wirkung. Aluminiumhydroxyd und DEAE-Dextran zeigten an nichtsensibilisierten Meerschweinchen eine stark toxische, komplettes Freund'sches Adjuvans dagegen eine schwach toxische Wirkung. In sensibilisierten Meerschweinchen fuhrte komplettes Freund'sches Adjuvans zur Immunzellbildung, die sich mit Makrophagen passiv ubertragen liesen. Summary Further Investigations on FMD-Allergy of the Delayed Type Comparative investigations on Frenkel- and BHK-allergic cattle by means of the CA-Test verified that an allergy of the delayed type as appears after repeated vaccinations is due to the FMD-virus alone as well as to FMD-virus in combination with allergenic BHK-cell components. The FMD-virus alone gives only a weak reaction. FMD-vaccines, containing allergenic BHK-cell components, cause the virus allergen to react more strongly in a sort of adjuvant effect. The virus RNA, obtained by splitting the virus, appeared not to be allergenic. The isolated virus-protein however had a strong allergenic action. Tests with sensitized guinea pigs supported this observation. Guinea pigs could be sensitized with the complete, purified FMD-virus as well as with split FMD-virus-protein or with BHK-cell substances. This allergy can be transmitted by macrophages. Generally the FMD-allergy was weaker than the BHK-allergy. An exception was in guinea pigs which were sensitized by a highly purified and concentrated virus pellet. In these animals in the CA-Test the reaction was almost entirely virus specific. It was not possible to sensitize the guinea pigs with pure virus-RNA. On the other hand the protein obtained from intensively purified FMD-virus caused a much higher degree of sensitization in guinea pigs than the complete virus. In the cross-CA-Test with guinea pigs sensitized against the virus types O1, A5, or C1, types A5 and C1 had a stronger reaction with one another than with O1. This type, however, had the strongest sensitizing effect. Aluminium hydroxide and DEAE dextran appeared to be highly toxic for non-sensitized guinea pigs, whereas Freund adjuvant, in contrast, was only slightly toxic. It appeared that in sensitized guinea pigs complete Freund adjuvant caused the production of immune cells which were transmissible passively by macrophages. Resume Suite des recherches sur la fievre aphteuse-allergie cellulaire Des analyses comparees a l'aide du test CA sur les boeufs allergiques au Frenkel et BHK confirment que, pour une allergie cellulaire (delayed type) apres une vaccination repetee, le virus-FA seul ainsi que combine avec des elements cellulaires-allergeniques-BHK provoque l'allergie. Le virus-FA seul n'a qu'un faible effet allergenique. Si les vaccines-FA contiennent des elements cellulaires-allergeniques-BHK ils amenent le virus-allergen a une reaction plus forte dans le sens d'un effet-adjuvant. La ribonucleine acide, gagnee par la division du virus n'a pas agie d'une facon allergenique. Par contre la proteine isolee avait un fort effet allergenique. Experiences servant d'exemple sur des cobayes sensibilises constaterent cette observation. Des cobayes purent ětre sensibilises non seulment par le virus-FA complet et depure ainsi que par la proteine divise du virus-FA, mais encore avec des elements cellulaires-BHK. Cet allergie se transmit grace aux macrophages. En regle l'allergie-FA avait une constitution plus faible que l'allergie-BHK. A l'exception des cobayes qui avaient ete sensibilises par le virus-pellet intensivement depure et concentre. Dans le cas present n'apparurent pratiquement que des reactions de virus specifique. Les cobayes ne purent ětre senibilises par l'acide Ribonucleine pure du virus. La proteine, separee du virus-FA fortement depuree avait sensibilise les cobayes avec plus d'intensite que le virus complet. Au test CA de croisement avec de cobayes qui etaient sensibilises contre les virus de type O1, A5, ou C1, les types A5 et C1 reagirent ensemble plus forte au test-CA qu'avec le type O1. Ce type, par contre, avait le plus fort de sensibilisation. Aluminiumhydroxyd et DEAE-dextrane avaint montre un fort effet toxique sur des cobayes non sensibilises, par contre, le Freund adjuvant avaient montre un faible effet toxique. Il apparait donc des cobayes sensibilises que le Freund adjuvant mene a la formation de cellules immunisees (immun-cells) qui etaient passivement transmissibles par les macrophages. Resumen Nuevas investigaciones sobre la alergia aftosa del tipo diferido Los estudios comparativos con ayuda de la prueba de anafilaxia cutanea (AC) en reses bovinas alergicas frente a la vacuna segun Frenkel o la elaborada en celulas renales de hamsters lactantes (BHK) confirman que la alergia del tipo diferido, registrada a continuacion de vacunaciones repetidas contra la glosopeda, se puede deber tanto al virus aftoso solo como en combinacion con los componentes celulares BHK alergenos. El virus aftoso solo no tiene mas que una accion alergena muy debil. Cuando las vacunas antiaftosas contienen fracciones celulares BHK alergenas, llevan al alergeno del virus a una reaccion mas intensa en el sentido de un efecto coadyuvante. El acido ribonucleico obtenido por desdoblamiento del virus no era alergeno, mientras que la proteina aislada del virus poseia una accion alergena muy marcada. Las observaciones experimentales en cobayas sensibilizados confirman este lance. Los cobayas se pudieron sensibilizar con el virus aftoso completo y purificado, con proteina disociada del mismo virus y tambien con substancias celulares BHK. La alergia pudo transmitirse a traves de macrofagos. Por regla general, la reaccion alergica aftosa se manifestaba de forma mas debil que la alergia BHK. Una excepcion la constituyeron los cobayas que previamente se sensibilizaron con concentrados de virus altamente purificados en la ultracentrifuga. En la prueba de AC no se obtuvieron practicamente mas que reacciones especificas de virus. Con acido ribonucleico puro del virus no resulto posible sensibilizar cobayas. La proteina disociada a partir de un virus aftoso harto purificado sensibilizaba los cobayas de manera mas efectiva que el virus completo. En la prueba cruzada de AC en cobayas, los cuales habian sido sensibilizados frente a los tipos de virus O1, A5 o C1, el A5 y C1 reaccionaban entre si con intensidad mayor que con O1. Pero este ultimo tipo poseia el efecto sensibilizante mas intenso. El hidroxido de aluminio y el DEAE-dextrano manifestaban en cobayas no sensibilizados una accion toxica fuerte, a la vez que el coadyuvante completo de Freund una accion toxica debil. En cobayas sensibilizados, el coadyuvante completo de Freund ocasionaba la formacion de celulas inmunes, las cuales se podian transmitir pasivamente por medio de los macrofagos.

Summary Five groups and one control group of 122 cows and heifers were parenterally vaccinated twice with different E. coli K 99 pilus vaccines at the end of gestation in order to increase and prolong K 99 pilus antibody secretion in milk. An ELISA was used to detect antibodies to K 99 pilus in milk. 83% control cows had antibodies to K 99 in colostrum. The average titer of colostrum K 99 pilus antibodies in non-vaccinated cows was 1:21. On the second day of lactation only small quantities of antibodies to K 99 pilus could be detected in the milk of 70 % of control cows. On the third day of lactation small quantities of antibodies were found in the milk of only 30% of control calves. Milk samples of the control group tested from the fourth day onwards did not show any K 99 pilus antibody activity. K 99 pilus antibody secretion in milk could be increased significantly by vaccinating the dams twice. The efficacy of vaccination was dependent on the adjuvant. Highest K 99 pilus antibody titers and the longest lasting secretion of antibodies to K 99 pilus were obtained with a vaccine containing saponin and aluminium hydroxide as adjuvant. The colostrum antibody titer to K 99 pilus antigen in this group was more than twelve times higher compared to the control group and was about eight times higher than the titer of the group which was given K 99 pilus antigen without adjuvant. On the 5th day of lactation, all vaccinated cows still had K 99 pilus antibodies in their milk. 80% and 64% of the vaccinated cows had antibodies to K 99 pilus in their milk on the 6th and 7th days of lactation respectively. Zusammenfassung Vakzinierung trachtiger Kuhe mit K 99- und Rotavirus-Antigenen: Wirksamkeit von K 99-Antigen und verschiedener Adjuvantien zur Stimulierung von Milchantikorpern 122 Kuhe und Farsen in funf Gruppen sowie einer Kontrollgruppe wurden gegen Ende der Trachtigkeit zweimal mit verschiedenen E. coli K 99 Pilus-Impfstoffen vakziniert, um die Ausscheidung von Antikorpern gegen K 99-Pili in der Milch zu erhohen und zu verlangern. 83% der Kontrollkuhe wiesen Antikorper gegen K 99 im Kolostrum auf, der Titer war im Durchschnitt 1:21. Am zweiten Laktationstag konnten nur mehr bei 70% der Kontrolltiere niedrige Antikorpertiter gegen K 99-Pili gefunden werden, am dritten Tag nur mehr bei 30%. Ab dem vierten Laktationstag wies keine der untersuchten Milchproben von Kontrollkuhen mesbare Antikorper gegen K 99-Pili auf. Die Ausscheidung von Antikorpern gegen K 99 in der Milch konnte durch zweimalige Vakzinierung der Tiere signifikant erhoht werden. Die Wirksamkeit der Impfung war abhangig vom verwendeten Adjuvans. Bei Verwendung einer Vakzine mit Saponin und Aluminiumhydroxid als Adjuvantien wurden die hochsten Antikorpertiter in der Milch und die langste Ausscheidungsdauer erzielt. In dieser Gruppe waren die Antikorpertiter gegen K 99-Pili zwolfmal hoher als in der Kontrollgruppe und achtmal hoher als bei Tieren, die K 99 ohne Adjuvans erhalten hatten. Am funften Laktationstag besasen noch alle geimpften Tiere Antikorper gegen K 99-Pili in der Milch, am sechsten und siebten Tag noch 80% bzw. 64%. Resume Vaccination de vaches avec un antigene K 99 et un antigene Rotavirus: pouvoir d'un antigene K 99 combine a differents adjuvants pour stimuler une secretion d'anticorps dans le lait 122 vaches et genisses reparties en 5 groupes et un groupe de controle ont ete vaccinees deux fois en fin de gestation avec differents vaccins de E. coli K 99-Pilus pour augmenter et prolonger l'excretion des anticorps contre K 99-Pili dans le lait. 83% de vaches de controle ont presente des anticorps contre K 99 dans le colostrum et le titre fut en moyenne de 1: 21. Des titres bas d'ancitorps contre K 99-Pili n'ont ete trouves au deuxieme jour de la lactation que chez le 70% des animaux de controle et seulement chez 30% au troisieme jour. Aucun anticorps mesurable contre K 99-Pili n'a ete trouve le quatrieme jour dans les echantillons de lait examines. L'excretion d'anticorps contre K 99 dans le lait a pu etre significativement augmentee par une vaccination des animaux en deux temps. L'efficacite de la vaccination a dependu de l'adjuvant utilise. On a obtenu les titres d'anticorps les plus eleves dans le lait et la duree d'excretion la plus longue en utilisant un vaccin avec de la saponine et de l'hydroxyde d'aluminium comme adjuvants. Les titres des anticorps et de l'hydroxyde d'aluminium comme adjuvants. Les titres des anticorps contre K 99-Pili dans ce groupe furent douze fois plus eleves que dans les groupes de controle et huit fois plus eleves que chez les animaux qui avaient recu K 99 sans adjuvant. Tous les animaux vaccines possedaient encore des anticorps contre K 99-Pili dans le lait au cinquieme jour de la lactation, le 80% au sixieme jour et le 64% au septieme jour. Resumen La vacunacion de vacas con antigenos K 99 y de Rotavirus: la eficacia del antigeno K 99 combinado con diferentes adyuvantes para estimular la secrecion de anticuerpos en la leche 122 vacas y vaquillonas, distribuidas en cinco grupos mas un grupo de testigos, fueron vacunadas dos veces hacia finales de la gestacion con diferentes vacunas flagelares E. coli K 99 para aumentar y prolongar la excrecion de anticuerpos anti K 99 flagelares en la leche. El 83% de las vacas testigo ostentaban anticuerpos anti K 99 en el calostro, siendo el titulo medio de 1: 21. El dia segundo de lactacion solo se pudieron hallar en 70% de los animates testigos titulos bajos de anticuerpos frente a los K 99 flagelares, mientras que el dia tercero solamente en 30%. A partir del dia cuarto de lactacion ninguna de las muestras de leche examinadas de las vacas de control presentaba anticuerpos mensurables antiflagelares K 99. La excrecion de anticuerpos anti K 99 con la leche se pudo aumentar significantemente por medio de la vacunacion doble de los animales. La eficacia de vacunacion dependia del adyuvante utilizado. Empleando una vacuna con saponina e hidroxido de aluminio como adyuvantes, se lograron los titulos maximos de anticuerpos en leche y la duracion maxima de excrecion. En este grupo eran doce veces mayores los titulos de anticuerpos antiflagelares K 99 que en el grupo testigo y ocho veces mayores que en los animales que habian recibido K 99 sin adyuvante. El dia quinto de lactacion poseian aun todos los animales vacunados anticuerpos antiflagelares K 99 en la leche, los dias sexto y septimo aun el 80% y 64%, resp. de los mismos.

Zusammenfassung Im Infektionsbelastungsversuch beim Kalb wurde vergleichend die orale Immunisierung gegen S. typhimurium mit hitzeinaktivierten Keimen and vermehrungsfahigen Gal E-Keimen untersucht. Die Beurteilung der Wirksamkeit erfolgte aufgrund des klinischen Verlaufes und Letalitat bei den Impflingen im Vergleich zu den nicht geimpften Kontrolltieren. Eine zehnmalige orale Impfung mit 1,2 · 1011 hitzeinaktivierten S. typhimurium-Keimen verlieh Kalber einen unvollstandigen Schutz gegen eine orale S. typhimurium-Belastunesinfektion. Unterschiede zwischen geimpften und nicht geimpften Tieren liesen sich in der Uberlebenszeit und Letalitatsrate, nicht aber im klinischen Erscheinungsbild nachweisen. Die Impfung wurde reaktionslos vertragen. Eine einmalige orale Impfung mit 1,2 · 1010 vermehrungsfahigen Gal E-S. typhimurium-Keimen schutzte Kalber gegen eine todliche Belastungsinfektion. Bei den Impflingen kam es aber zu schweren Impferkrankungen (Enteritis, Fieber). Wurden die Impfdosen gesenkt, gingen zwar die Impferkrankungen zuruck, der Impfschutz war dafur aber nicht starker belastungsfahig als nach Schutzimpfung mit inaktivierten Keimen. Summary Comparative investigations on oral immunization with heat-inactivated and live, avirulent (gal E-) S. typhimurium bacteria against salmonellosis in calves In calves, the efficacy of oral immunization against S. typhimurium with heat-inactivated bacteria and live gal E-mutants was investigated by challenge experiments. Clinical signs and survival rates were compared between vaccinated and non-vaccinated animals in order to determine the efficacy of immunization. Ten oral doses of 1.2 · 1011 heat-inactivated S. typhimurium bacteria induced only incomplete protection against oral S. typhimurium-test infection in calves. Differences between vaccinated and non-vaccinated calves were observed only with respect to survival rates but not to clinical signs and course. Vaccination with heat-inactivated bacteria caused no side-effects. A single oral immunization with 1.2 · 1010 live gal E-S. typhimurium protected calves completely against a lethal dose of S. typhimurium. In vaccinates, however, application of gal E-mutants resulted in severe illness (enteritis, fever) in all animals. After lowering the vaccination dose of gal E-vaccine, side-effects subsided, but post-vaccinal immunity decreased. Resume Recherches comparees sur l'action d'une immunisation orale avec des germes de S. typhimurium (Gal E) thermoinactives mais se multipliant et avirulents contre la salmonellose du veau On a examine par comparaison l'immunisation orale contre S. typhimurium avec des germes thermoinactives et des germes multiplicables Gal E dans des essais d'infection chez le veau. L'estimation de l'efficacite a ete triee a partir de l'evolution clinique et de la letalite chez les animaux vaccines par rapport aux animaux de controle non vaccines. Une vaccination orale en dix fois avec 1,2 × 1011 germes thermoinactives de S. typhimurium a donne une protection incomplete aux veaux infectes oralement par S. typhimurium. Des differences entre animaux vaccines et non vaccines n'ont pas ete constatees dans le temps de survie et le taux de letalite mais ont ete observees dans les symptomes cliniques. La vaccination fut supportee sans reactions. Une vaccination orale unique avec 1,2 × 1010 germes multiplicables Gal E — S. typhimurium a protege des veaux contre une infection mortelle. Les animaux vaccines ont presente toutefois de graves lesions vaccinales (enterite, fievre). Lorsque la dose vaccinale fut diminuee, les lesions vaccinales ont diminue mais la protection ne fut pas plus grande que celle constatee avec des germes inactives. Resumen Estudios comparados sobre la eficacia de una inmunizacion oral con germenes (Gal E) S. typhimurium termoinactivados y multiplicables avirulentos contra la salmonelosis de las terneras En el ensayo de infeccion de desafio en la ternera se estudio comparativamente la inmunizacion oral contra S. typhimurium con germenes termoinactivados y germenes Gal E multiplicables. Para valorar la eficacia se recurrio al curso clinico y a la letalidad en los vacunados en comparacion con los de los animales testigos no vacunados. La vacunacion oral en diez ocasiones con 1.2 × 1011 germenes S. typhimurium termoinactivados proporciono a las terneras una proteccion incompleta frente a una infeccion oral de desafio con S. typhimurium. Las diferencias entre los animales vacunados y los que no lo estaban se pudieron poner en evidencia en el tiempo de supervivencia y en la tasa de letalidad, pero no mediante el cuadro sintomatico clinico. La vacunacion fue tolerada sin reaccion alguna. Una vacunacion oral unica con 1.2 × 1010 germenes Gal E-S. typhimurium multiplicables protegio a las terneras frente a una infeccion de desafio mortal. Pero en los animales vacunados se presentaron enfermedades vacunales graves, tales como enteritis y fiebre. Al reducir las dosis de vacunacion, es cierto que se redujeron las enfermedades vacunales, pero para eso la proteccion de la vacuna no se pudo gravar mas que tras la vacunacion protectora co*** germenes inactivados.

Zusammenfassung Neugeborene Kalber wurden in den ersten 10 Lebenstagen mit einer aus hitzeinaktivierten E. coli-Bakterien bestehenden, mono- oder polyvalenten Vakzine oral immunisiert. Die Gesamtimpfdosis betrug 1011 inaktivierte Keime. Die Wirksamkeit pruften wir experimentell mit dem Darmligaturtest und in einem uber 31/2 Jahre laufenden Feldversuch. Bereits 1 Tag nach der letzten Impfung war eine gut belastbare lokale Darmimmunitat nachweisbar. In 38 Problembestanden mit 1500 Tieren lies sich mit der stallspezifischen Schluckimpfung die durchschnittliche Mortalitat von 24 Prozent auf unter 1 Prozent senken. Mit der polyvalenten Vakzine, die die O-Antigene 78, 86, 35, 8 und 117 enthielt, hatten wir nach Impfung von 1000 Tieren in 72 Prozent der Falle Erfolg. Der Schutz erstreckte sich stets nur auf die in der Vakzine enthaltenen Serotypen. Summary Oral active immunisation of new-born calves against E. coli: Efficiency test using intestinal ligature test and field trial Newborn calves were vaccinated orally during the first 10 days of life with a mono- or polyvalent vaccine consisting of heat-inactivated E. coli-antigens. The total vaccine-dose was 1011 inactivated organisms. Efficiency was tested by the intestinal ligature test and field trial over 31/2 years. Already 1 day after the last vaccination there was a well challengeable local intestinal immunity. In 38 “problem-herds” with a total of 1,500 animals the mortality rate of 24% was reduced to 1% after administering a herd-specific oral vaccine. The polyvalent vaccine, containing the O antigens 78, 86, 35, 8 and 117 was successful in 72% out of 1,000 animals vaccinated. Protection was effective only against the serotypes contained in the vaccine. Resume Immunisation orale active de veaux nouveau-nes contre E. coli: preuve de l'efficacite dans le test de ligature intestinale et dans le terrain Des veaux noveau-nes ont ete immunises dans les 10 premiers jours de vie avec un vaccin oral mono- ou polyvalent d'E. coli inactive a la chaleur. La dose vaccinale comportait 1011 germes inactives. On a teste l'efficacite experimentalement avec le test de ligature intestinale et dans un essai sur le terrain durant 31/2 ans. On a mis en evidence une bonne immunite intestinale locale 1 jour apres la derniere vaccination. Dans 38 explotations a problemes avec 1500 animaux, la vaccination orale specifique d'etable a reduit la mortalite moyenne de 24 a 1%. Apres une vaccination de 1000 animaux avec des vaccins polyvalents contenant les antigenes 078, 086, 036, 08 et 0117, on a obtenu un 72% de succes. La protection ne s'etendait qu'aux serotypes contenus dans le vaccin. Resumen Inmunizacion oral activa de terneros recien nacidos contra E. coli: contraste de la eficacia en la prueba de la ligadura intestinal y en el ensayo campal Se inmunizaron terneros recien nacidos por via oral con una vacuna mono o polivalente que consistia en bacterias E. coli termoinactivadas. La dosis total de vacunacion era del orden de 1011 germenes inactivados. Contrastamos la eficacia experimentalmente con la prueba de ligadura intestinal y en un ensayo campal de 3 anos y 1/2 de duracion. Al cabo de un dia nada mas despues de la vacunacion ultima se podia identificar una inmunidad intestinal local gravable buena. En 38 explotaciones problematicas con 1500 animales se logro reducir con la vacunacion bebible, especifica de establo, la mortalidad promedio de 24 a menos de 1%. Con la vacuna polivalente, la cual contenia los antigenos 0 78, 86, 35, 8 y 117, tuvimos exito en el 72% de los casos despues de haber vacunado 1000 animales. La proteccion solo era efectiva frente a los serotipos contenidos en la vacuna.

Zusammenfassung Neugeborene Ferkel wurden an den ersten zehn Lebenstagen einmal taglich oral gegen E. coli geimpft. Die Schluckvakzine bestand aus hitzeinaktiviertem E. coli 08-Vollantigen. Am 14. Tag nach der letzten Immunisierung wurde die Wirksamkeit mit dem Darmligaturtest gepruft. Geimpfte Ferkel konnten bei gleichem Ergebnis im Darmligaturtest mit einer 100- bis 1000fach hoheren Keimzahl belastet werden als nicht geimpfte Kontrolltiere. Summary Active, oral immunisation of new-born piglets against E. coli: Efficiency tests using the intestinal ligature test New-born piglets were immunised orally once daily during their first ten days of life against E. coli. The oral vaccine was prepared from heat inactivated E. coli 08 complete antigen. The efficiency of the vaccine was examined in the intestinal ligature test 14 days after the last immunisation. Results of challenge using the intestinal ligature test showed that vaccinated piglets resisted a 100 to 1,000 times higher dose of E. coli than non-vaccinated control animals. Resume Immunisation orale active de porcelets nouveau-nes contre E. coli: preuve de l'efficacite dans le test de ligature de l'intestin Des porcelets nouveau-nes ont ete vaccines oralement une fois par jour pendant les dix premiers jours de vie contre E. coli. Le vaccin oral se composait de l'antigene complet 08 d'E. coli inactive par la chaleur. L'efficacite fut eprouve epar le test de ligature de l'intestin au quatorzieme jour suivant la derniere immunisation. Les porcelets vaccines donnant les memes resultats au test de ligature ont supporte un nombre de germes de 100 a 1000 fois plus eleve que les animaux de controle non vaccines. Resumen Inmunizacion oral activa de lechones recien nacidos frente a Escherichia coli: demostracion de eficacia en la prueba de ligadura intestinal Lechones recien nacidos se vacunaron durante los diez primeros dias de vida, una vez por jornada, por via oral contra E. coli. La vacuna bebible consistia en antigeno total E. coli 08 termoinactivado. El dia 14° tras la inmunizacion ultima se contrasto la eficacia con la prueba de intestino ligado. Los lechones vacunados se podian gravar, siendo el resultado similar en la prueba de intestino ligado, con una cantidad de germenes de 100 a 1000 veces mayor que los animales testigos no vacunados.

Zusammenfassung Kotproben von 100 neugeborenen Kalbern mit Diarrhoen unterschiedlicher Dauer enthielten ausnahmslos E. coli-Bakterien. Bei 40 Kotproben wurden gleichzeitig Rotaviren nachgewiesen. Die isolierten E. coli-Stamme wurden im Darmligaturtest beim Ferkel auf Enterotoxinbildung sowie mit Hilfe der Objekttragerschnellagglutination bzw. dem Hamagglutinationstest auf O- und K (Fimbrien)-Antigene untersucht. 54 E. coli-Stamme bildeten regelmasig Enterotoxin, von diesen 54 Stammen wiesen 37 das K 99-Antigen auf, 42 Stamme zeigten eine mannose-resistente Hamagglutination von Pferdeerythrozyten. Bei den 54 Tieren, bei denen enterotoxinbildende E. coli nachweisbar waren, betrug der Anteil der Rotavirus-positiven Proben 43% (23 Tiere). Von 63 K 99-negativen Stammen bildeten 17 regelmasig und 36 unregelmasig Enterotoxin, 5 Stamme zeigten eine mannose-resistente Hamagglutination. Hier wiesen 25 Proben Rotaviren auf (40%). Die 0-Gruppe 106 dominierte bei den enteropathogenen E. coli-Stammen. Summary Demonstration of enteropathogenic Escherichia coli-strains and rotaviruses in faecal samples from calves with diarrhoea Faecal samples collected from 100 neonatal calves with diarrhoea of variable duration all contained E. coli bacteria. Rotaviruses were demonstrated in 40 samples at the same time. The isolated E. coli strains were tested for their ability to produce enterotoxin by using the intestinal ligature test, and for the presence of O and K (pili) antigens by employing slide agglutination and haemagglutination techniques. 54 E. coli strains regularly produced enterotoxin, 37 of these 54 strains contained K 99 antigens, 42 strains showed a mannose-resistent haemagglutination of horse red blood cells. The number of rotavirus-positive samples in the enterotoxin-producing group was 23 (43%); of the 63 K 99-negative strains, 17 produced enterotoxin regularly and 36 produced it irregularly, 5 strains showed a mannose-resistent haemagglutination. 25 samples were positive for rotavirus (40%). 0 group 106 dominated within the enteropathogenic E. coli strains. Resume Mise en evidence de souches enteropathogenes d'E. coli et de Rotavirus dans des matieres fecales de veaux atteints de diarrhee Les echantillons de matieres fecales de 100 veaux nouveau-nes atteints de diarrhee de duree variable contenaient sans exception des bacteries E. coli. Des Rotavirus ont ete mis en evidence simultanement dans 40 echantillons de feces. Les souches isolees de E. coli ont ete examinees du point de vue de la formation d'enterotoxine dans le test de ligature de l'intestin chez le porcelet et pour la recherche des antigenes O et K (pili) au moyen de l'agglutination rapide sur lame et du test d'hemagglutination. 54 souches d'E. coli ont regulierement produit une enterotoxine; 37 de ces 54 souches possedaient l'antigene K 99 et 42 souches presenterent une hemagglutination mannose-resistante des globules rouges de cheval. Sur les 54 animaux chez lesquels un E. coli formant une toxine fut mis en evidence 43% des echantillons de matieres fecales furent Rotavirus-positifs (23 animaux). Sur les 63 souches K 99 negatives, 17 formeret regulierement une enterotoxine et 36 irregulierement, 5 souches montrant une hemagglutination mannose-resistante. 25 echantillons presenterent dans ce cas du Rotavirus (40%). Le groupe O/106 a domine dans les souches enteropathogenes d'E. coli. Resumen Identificacion de estirpes enteropatogenas de Escherichia coli y de Rotavirus en las materias fecales de terneros con diarrea Las muestras de materias fecales de 100 terneros recien nacidos con diarrea contenian sin excepcion bacterias E. coli. El Rotavirus se puso en evidencia simultaneamente en 40 muestras de heces. Las cepas aisladas de E. coli se examinaron bajo el punto de vista de la formacion de enterotoxina en la prueba de ligadura del intestino en el lechon y mediante la pesquisa de antigenos O y K (fimbriales) con ayuda de la aglutinacion rapida sobre portaobjetos resp. la prueba de hemoaglutinacion. 54 estirpes de E. coli formaban regularmente enterotoxina; 37 de estas 54 estirpes poseian el antigeno K 99 y 42 cepas presentaban una hemoaglutinacion manosarresistente de los globulos rojos de caballo. En los 54 animales, en los que se identifico E. coli enterotoxinogena, el 43% de las muestras eran Rotavirus-positivas (23 animales). De las 63 cepas K 99-negativas formaban 17 regularmente y 36 irregularmente enterotoxina, 5 estirpes mostraban una hemoaglutinacion manosarresistente. En este caso mostraban 25 muestras Rotavirus (40%). El grupo 0/106 dominaba en las estirpes E. coli enteropatogenas.

Zusammenfassung In der Zeit von 1967-70 wurden in Bayern 15 485 832 Rinder gegen die Maul- und Klauenseuche schutzgeimpft. Dabei wurden 3052 postvaccinale Storungen der Trachtigkeit beobachtet und statistisch erfast. Die Schadensquote verteilt sich in etwa gleichmasig auf alle 4 in Bayern in diesem Zeitraum eingesetzten Impfstoffarten (zwei Frenkel-, eine BHK- und eine Kalbernierenkultur-Vaccine). Die Ursache von postvaccinalen Verwerfensfallen ist unbekannt. Fur die Diagnose stellt deshalb der Zeitfaktor das wichtigste Kriterium dar. Bei Trachtigkeitsstorungen, die 10 Stunden bis 14 Tage nach einer MKS-Schutzimpfung auftreten, sollte die ursachliche Beteiligung mit der Impfung anerkannt werden, sofern spezifische Ursachen, wie z. B. Deckseuchenerreger, ausgeschlossen werden konnen. Eine Beziehung zwischen den postvaccinalen Allergien vom Spattyp und den Trachtigkeitsstorungen lies sich nicht nachweisen. Ahnlichkeiten im Haufigkeitstrend bestanden dagegen zwischen den postvaccinalen Allergien vom Fruhtyp und den Trachtigkeitsstorungen. Bei zunehmender Zahl von Revaccinationen steigt die Quote von Trachtigkeitsstorungen an. Besonders belastet ist der Zeitraum zwischen 6. und 9. Trachtigkeitsmonat. Summary Statistical studies on disturbances in pregnancy which followed the vaccination campaigns against foot-and-mouth disease in Bavaria in 1967-70 During the period 1967-70 a total of 15,485,832 cattle were vaccinated against foot-and-mouth disease. Among them there were 3,052 cases of postvaccinal disturbance of pregnancy observed and statistically examined. The number of affected animals was fairly equally distributed between the four types of vaccine used - two Frenkel, one BHK and one calf kidney culture vaccine. The cause of the post-vaccinal abortions and premature births is not known. For diagnosis the most important criterion is the time factor. Disturbances of pregnancy that take place between 6 hours and 14 days after vaccination against F and M can be considered associated with the vaccination if specific causes, such as genital tract infection, can be excluded. A relationship between post-vaccinal allergy of the delayed type and disturbances of pregnancy could not be demonstrated. On the other hand similarities in the frequency trend between post-vaccinal allergy of the early type and disturbance of pregnancy existed. With an increasing number of revaccinations the incidence of pregnancy disturbances rose. Particular susceptibility existed between the 6th and 9th months of pregnancy. Resume Recherdies statistiques sur les troubles de la gestation parvenus a la suite de la vaccination contre la fievre aphteuse de 1967–1970 en Baviere De 1967 a 1970, on a procede en Baviere a des vaccinations contre la fievre aphteuse sur 15 485 832 bovins. Par la suite,3 052 cas not manifeste des troubles post-vaccinaux de la gestation, qu'on a evalues statistiquement. La repartition des dommages est a peu pres egale avec les quatre sortes de vaccins appliquees en Baviere a cette epoque (deux vaccins Frenkel, un BHK et un vaccin cultive sur reins de veaux.) La cause de ces avortements post-vaccinaux est inconnue. Pour le diagnostic, c'est le facteur temps qui represente le critere le plus important. Lors de troubles de la gestation qui apparaissent 10 heures a 14 jours apres une vaccination contre la fievre aphteuse, il faut admettre une participation causale de la vaccination, si des causes specifiques, telles qu'un agent d'une maladie venerienne, sont exclues. On ne peut etablir aucune relation entre les allergies post-vaccinales du type retarde et les troubles de la gestation. Par contre, on trouve des ressemblances dans la frequence des allergies post-vaccinales du typ immediat et des troubles de la gestation. La frequence des troubles de la gestation augmente avec le nombre des revaccinations. La periode la plus exposee s'etend du 6eme au 9eme mois de la gestation. Resumen Investigaciones estadisticas sobre los trastornos de gestacion que surgieron por efecto de la vacunacion preventia antiaftosa en Baviera en los anos 1967–1970 En el periodo de 1967–1970 en Baviera fueron vacunados preventivamente contra fiebre aftosa 15.485.832 cabezas de ganado vacuno. Se observaron 3.052 casos de trastornos de gestacion post-vacunales, los cuales fueron registrados estadisticamente. El numero de danos se distribuye en proporcion casi simetrica entre los cuatro tipos de vacuna empleados en Baviera en este lapso (dos del tipo Frenkel-, una del tipo BHK-, y una vacuna de cultura de rinon de ternero). La causa de los abortos post-vacunales es desconocida. Por esto representa para el diagnostico el factor tiempo el criterio mas importante. En trastornos de gestacion que se presentan de 10 horas hasta 14 dias despues de una vacunacion preventiva antiaftosa se debria reconocer la participacion causal de la vacuna, ya que se pueden eliminar causas especificas, como por ejemplo infecciones transmitidas durante la monta. No se pudo comprobar una relacion entre las alergias post-vacunales de tipo diferido (delayed type) y los trastornos de gestacion. Existe en cambio una semejanza en la frecuencia entre las alergias postvacunales de tipo inmediato y los trastornos de gestacion. Con el aumento del numero de revacunaciones crece el numero de trastornos de gestacion. Esto afecta particularmente el periodo del 6 ° al 9 ° mes de prenez.

Zusammenfassung Die im Rahmen der Landesimpfungen gegen die MKS in Bayern in den Jahren 1967/68 aufgetretenen Impfkomplikationen lassen sich in 3 Gruppen aufgliedern: 1. in Impferkrankungen, 2. in Impfdurchbruche und 3. in Impfschaden. Die Impfkomplikationen der 1. und 2. Gruppe haben die Impfungen nicht belastet. Anders verhielt es sich mit den Impfschaden. Hier dominierten die Allergien mit den sich darin verbergenden anaphylaktoiden Reaktionen und die Storungen der Trachtigkeit. Die statistische Auswertung der Impfkomplikationen bezog sich 1967 auf 3 021 758 Tiere mit BHK-Vaccinen und 1 192 502 Tiere mit Frenkel-Vaccinen, 1968 auf 3 093 650 Tiere mit BHK-Vaccinen und 1 169 084 Tiere mit Frenke-Vaccinen. Beide Impfstoffarten waren gemeinsam belastet durch Allergien vom Soforttyp (immediate type), unter denen sich anaphylaktoide Reaktionen verbergen konnten, durch Storungen der Trachtigkeit, durch Lokalreaktionen und durch sonstige, unspezifische Schaden. Einseitig belastet waren demgegenuber die BHK-Vaccinen. Hier dominierten als Impfschaden die Allergien vom Spattyp (delayed type). Sie stiegen bei Revaccination signifikant an. Die Ursachen der einzelnen Impfkomplikationen werden aufgezeigt und hieraus Schlusfolgerungen fur die Herstellung der MKS-Vaccinen gezogen. Zur Vermeidung von Protein- und MKS-Antigen-Allergien, die sich als wichtigste Ursache der Impfkomplikationen herausstellen, mussen MKS-Vaccinen, die fur eine sich standig wiederholende Landesimpfung eingesetzt werden, bezuglich Eiweis, das nicht an den immunisierenden MKS-Antigen-Komplex gebunden ist, gereinigt werden. Dies betrifft vor allem Vaccinen, die aus nicht fur das Rind homologen Zellsystemen hergestellt werden. Zudem sollte Formalin durch Inaktivierungsmittel ersetzt werden, die homologes Rindereiweis nicht denaturieren. Summary Studies on the nature, extent and causation of vaccination complications after the foot-and-mouth disease vaccination campaign of 1967/8 in Bavaria The vaccination complications in this campaign are classified into 3 groups: — 1. Vaccination causing foot-and-mouth disease. 2. Vaccination breakdowns. 3. Vaccination illness. The first two were not serious for the animal, whereas the third kind was dominated by masked allergic reactions and interference with pregnancy. Statistical evaluation of vaccination complications was made in 1967 on 3,021,758 animals given BHK vaccines and 1,192,502 given Frenkel vaccine: for 1968 the numbers were 3,093,650 and 1,169,084 cattle, respectively. Both types of vaccine were associated with allergic reactions of the immediate kind, characterised by anaphylactoid reactions, interference with pregnancy, local reactions and various non-specific injuries. In contrast, the BHK vaccines alone gave allergic reactions of the delayed type which became more marked at revaccination. The causes of the individual vaccination complications were examined and their relevance to F and M vaccine production is discussed. To avoid protein and F and M allergens, which are the most important cause of vaccination complications, F and M vaccines for repeated use in the field must be freed from proteins that are not associated with the F and M antigens. This applies to all vaccines which are not prepared from cell systems homologous with cattle. Formalin should be replaced by an agent that does not denature ox protein. Resume Recherches sur le genre, l'importance et les causes des dommages dus aux vaccinations protectrices contre la fievre aphteuse en Baviere dans les annees 1967/68 Les complications qui se sont produites en Baviere a la suite des vaccinations nationales contre la fievre aphteuse dans les annees 1967/68 peuvent etre reparties en trois groupes: 1. reactions generales, 2. reactions locales et 3. complications reelles. Les complications de la vaccination du ler et du 2eme groupe sont sans consequences graves. Mais il en est tout autrement avec les complications de 3eme groupe, ou l'on trouve principalement des allergies, avec des reactions anaphylactoides masquees, et des troubles de la gestation. L'evaluation statistique des complications de la vaccination porte pour 1967 sur 3 021 758 animaux ayant recu le vaccin BHK et sur 1 192 502 animaux ayant recu le vaccin Frenkel et pour 1968 sur 3 093 650 animaux ayant recu le vaccin BHK et sur 1 169 084 animaux ayant recu le vaccin Frenkel. Les deux sortes de vaccins sont genees par des allergies du type immediat, pouvant dissimuler des reactions locales et par d'autres dommages non specifiques. Par contre, seuls les vaccins BHK sont genes par des complications de vaccination, telles que les allergies du typu retarde (delayed type), qui augmentent de facon significative lors d'une revaccination. Les causes de chaque sorte de complication sont indiquees et on en tire des conclusions pour la preparation des vaccins contre la fievre aphteuse. Pour eviter les allergies aux proteines et a l'antigene de la fievre aphteuse, qui s'averent etre la cause la plus importante des complications de la vaccination, il faut purifier les vaccins employes de facon repetee pour une vaccination nationale contre la fievre aphteuse, afin d'eliminer les proteines non liees au complexe immunisant de l'antigene de la fievre aphteuse. Ceci concerne surtout les vaccins qui sont prepares a partir de systemes cellulaires non homologues pour le boeuf. Il faudrait encore remplacer la formaline par un agent d'inactivation que ne denature pas les proteines homolngues de boeuf. Resumen Estudios sobre la naturaleza, extension y causas de los danos ocasionados por la vacunacion preventiva antiaftosa en Baviera durante los anos 1967/68 Las complicaciones de vacunacion que surgieron al margen de la campana antiaftosa estatal en Baviera en los anos 1967/68 se pueden dividir en 3 grupos: 1° en enfermedades derivadas de la vacunacion, 2° en perforaciones de la zona vacunal inmune y 3° en danos vacunales. Las complicaciones vacunales correspondientes a los grupos 1° y 2° no han gravado las vacunaciones efectuadas. Otro fue el resultado de los danos vacunales. Aqui dominaban las alergias con las reacciones anafilactoides ocultas en las mismas, y los trastornos de la gestacion. La evaluacion estadistica de las complicaciones de vacunacion se refirio en 1967 a 3.021.758 animales con vacunas BHK (cultivo celular) y 1.192.502 con vacunas del tipo Frenkel, mientras que en 1968 a 3.093.650 animales con vacunas BHK y 1.169.084 con las de tipo Frenkel. Ambos tipos de la vacuna estaban gravados conjuntamente por alergias de tipo inmediato, entre las que se pueden ocultar reacciones anafilactoides, por trastornos de la gestacion, reacciones locales y otros danos inespecificos. Sin embargo, gravadas de forma unilateral estaban las vacunas BHK. Aqui dominaban como danos de vacunacion las alergias de tipo diferido. Estas aumentaban de manera significativa en las revacunaciones. Se ilustran las causas de las complicaciones vacunales diversas, sacandose de ellas las conclusiones para la elaboracion de las vacunas contra la fiebre aftosa. Para impedir las alergias proteicas y aftosa-antigenicas, que resultaron ser la causa predominante de las complicaciones vacunales, tienen que purificarse las vacunas contra la glosopeda, que se vayan a emplear en las campanas estatales periodicas, en cuanto a los albuminoides que no esten ligados al complejo inmunizante del antigeno aftoso. Esto atane, ante todo, a las vacunas preparadas a base de sistemas celulares no homologos para el ganado vacuno. Ademas, deberea sustituirse el formol por substancias inactivantes, que no desnaturalizasen las proteinas bovinas homologas.

Zusammenfassung Gegenstand: Ziel der Untersuchung war, die Wirkung zweier Probiotika (Stammesvarianten von Bac. cereus) auf die Zusammensetzung der Darmflora und zootechnische Parameter beim früh entwöhnten Ferkel zu prüfen. Material und Methoden: Neben einer Ausgangsgruppe (Gruppe 0) wurde bei drei Versuchsgruppen am Ende eines 35-tägigen Stoffwechselversuchs die Darmflora untersucht. Die Ferkel wurden auf folgende Gruppen verteilt: Kontrollgruppe (Gruppe I), Paciflor-Gruppe (Bac. cereus var. caron, Gruppe II) und Toyocerin-Gruppe (Bac. cereus var. toyoi, Gruppe III). Die Zulage von Sporen des Bacillus cereus erfolgte jeweils in einer Dosierung von 109 cfu/kg zu einer Diät auf Mais-Soja-Basis. Ergebnisse: Die Gabe von Probiotika hatte weder auf die mittlere tägliche Lebendmassezunahme, bei insgesamt sehr guter Futterverwertung (1,7:1), noch auf die Gesamtfutteraufnahme einen statistisch signifikanten Einfluss. Bei den keimsupplementierten Tieren ließen sich am Ende des Versuchs in allen Darmsegmenten stabile Populationen an Bac. cereus nachweisen. Die additive Wirkung der veränderten Hygieneverhältnisse und die leicht restriktive Gabe der Diät bewirkten eine signifikante Reduktion coliformer Keime im Duodenum und Rektum bei Gabe beider Varianten von Bac. cereus. Ohne Probiotika- Einfluss, jedoch statistisch abgesichert, war die geringere Enterokokken-Konzentration im Zäkum und Rektum der Tiere aus den Gruppen I, II und III im Vergleich zu Gruppe 0. Im Duodenum ließen sich zu Versuchsende in allen drei Gruppen im Vergleich zur Ausgangsgruppe signifikant geringere Konzentrationen an coliformen Keimen nachweisen. In Gruppe III kam es zu einer signifikanten Steigerung der Laktobazillen-Konzentration. Die Anwendung von Toyocerin führte zu signifikant höheren Konzentrationen coliformer Keime im Zäkum. Unter Berücksichtigung der Keimkonzentration in der Ausgangsgruppe kann dies als florastabilisierende Wirkung von Toyocerin gedeutet werden. Schlussfolgerung und klinische Relevanz: Trotz der stabilisierenden Wirkung auf die Darmflora sowie zu erwartender positiver zootechnischer Effekte sollten beim Einsatz von Probiotika in der Ferkelaufzucht optimale Nährstoffversorgung und Hygienebedingungen nicht vernachlässigt werden.

ZusammenfassungIm Rahmen der vorliegenden Studie wurde eine repräsentative Stichprobe an Blutproben, die zwischen Februar und Juli 1998 von 1505 Warmblutstuten aus 415 verschiedenen Pferdehaltungen im Zuchtgebiet Westfalen-Lippe entnommen wurden, auf neutralisierende Antikörper gegen equines Arteritisvirus (EAV) untersucht. Insgesamt ergab sich eine Seroprävalenzrate des EAV von 27,6%. Die statistische Auswertung der einzelnen Ergebnisse nach Kriterien, die sich aus der Anamnese ergaben, erbrachte zum Teil signifikante Unterschiede in der Höhe der Seroprävalenzraten. Von einem schwach bis hoch signifikanten Einfluss auf die Rate seropositiver Tiere waren die Größe der Betriebe (Anzahl der Pferde) und das Ausmaß des Pferdeverkehrs, das Alter der Stuten, die Anzahl der bisherigen Trächtigkeiten sowie die zum Zeitpunkt der Untersuchung erfolgte Bedeckung/ Besamung und Trächtigkeit. Hierzu gehörten auch Ergebnisse aus anamnestischen Erhebungen über verzögerte Konzeptionen, mehrfaches »Umrossen«, Fruchtverluste sowie Frühgeburten und die Geburt lebensschwacher Fohlen. Eine Assoziation weiterer in die Untersuchung einbezogener Kriterien, insbesondere anamnestisch erhobener in der Vergangenheit aufgetretener Infektionskrankheiten, mit der Seroprävalenzrate konnte nicht hergestellt werden.

In this study, organ samples from 426 common seals Phoca vitulina, 298 harbour porpoises Phocoena phocoena, 34 grey seals Halichoerus grypus and 10 other marine mammals were assessed for the presence of Brucella species. Forty-seven common seals, 2 harbour porpoises and 1 grey seal were found to be positive for these bacteria. A total of 91 Brucella strains were successfully isolated, due to the fact that Brucella spp. were found in more than one organ sample in 15 animals. The primary organ in which the bacteria were present was the lung. In addition, 2 strains were isolated from lungworms (Parafilaroides spp.). Forty-nine of the isolated strains were selected for further analysis using conventional phenotyping methods. Molecular characterisation was carried out by analysing the IS711 and omp2 loci. With respect to the distribution of the IS711 loci in the genome, the 49 field isolates differed strongly from the terrestrial Brucella species and marginally from the marine Brucella reference strain NCTC12890. Based on the results of the PCR restriction fragment length polymorphism (PCR-RFLP) investigation of the omp2 locus, the majority of the Brucella field isolates were classified as B. pinnipediae, recently proposed B. pinnipedialis, possessing 1 omp2a gene and 1 omp2b gene. Two field isolates revealed the presence of 2 omp2a genes, as has been described for Brucella ovis. To our knowledge, these results confirm for the first time the presence of Brucella species in the marine mammal population of the German North Sea. These findings highlight the need for additional research on the relevance of these Brucella species for marine hosts and their environment.

Summary Experimental Studies Comparing the Immunostimulating (Paramunizing) Effect of Bacterial Lysates in Bacterial Infection Models With the Chromium 51 Test Demonstrating Spontaneous (Natural) Cytotoxicity Bacterial lysates of different bacterial strains (E. coli, B. bronchiseptica, P. haemolytica) were prepared by heating, acid- and alcaline-hydrolysis. Lysates were tested for their immunostimulating effect in bacterial infection models and with chromium 51 test demonstrating spontaneous (natural) cytotoxicity. Lysate production was standardized by protein- and Lps-determination. The alcaline-hydrolysis reduced toxicity of Lps and increased the content of soluble bacterial protein. Heating and acid-hydrolysis did not alter bacterial suspensions with respect to Lps-toxicity and protein-content. Mice infected with P. aeruginosa, P. multocida, E. coli and L. monozytogenes (5–10 LD50) had a significantly longer survival time after prophylactic immunostimulation with bacterial lysates than control animals. No protection was observed in immunostimulated mice infected with Erysipelothrix rhusiopathiae. In the Pseudomonas infection model, bacterial lysates prepared by alcaline-hydrolysis had a 10 times higher immunostimulating effect than lysates prepared by acid-hydrolysis or heating. Bacterial lysates stimulated spontaneous cytotoxicity of natural mouse peritoneal killer cells after intraperitoneal application. Whole bacterial lysates had a higher NK-activity as their corresponding purified lipopolysaccharide portion. Zusammenfassung Mit Hitze, Saure oder Lauge inaktivierte Keimsuspensionen verschiedener Bakterien (E. coli, B. bronchiseptica, P. haemolytica) wurden in bakteriellen Infektionsmodellen und im Cytotoxizitatstest auf ihre immunmodulierenden Wirkungen bei der Maus getestet. Die Standardisierung der verschiedenen Praparate erfolgte mittels Protein- und LPS-Bestimmung. Die alkalische Hydrolyse reduzierte im Vergleich zur unbehandelten Ausgangssuspension die Toxizitat des Lipopolysaccharids und fuhrte zur Freisetzung von bakteriellem Protein. Erhitzung oder saure Hydrolyse hatten keine mesbaren Veranderungen im Protein- oder LPS-Gehalt der Keimsuspensionen zur Folge. Die bakteriellen Praparate erhohten nach prophylaktischer intraperitonealer Applikation (24 h ante infect.) die Uberlebensrate der Mause gegen Belastungsinfektionen mit P. aeruginosa-, P. multocida-, E. coli- und L. monozytogenes-Keimen. Unwirksam blieben die Praparate im Rotlaufmodell. Das durch alkalische Hydrolyse hergestellte Praparat erzielte 10fach mehr Wirkungseinheiten (5000 WE) im Pseudomonas-Modell als das saure Hydrolysat oder das Hitzelysat. Im Cytotoxizitatstest waren keine Wirksamkeitsunterschiede festzustellen. Grundsatzlich hatte das „Gesamtpraparat” eine starkere Wirkung als der phenolextrahierte Lipopolysaccharidanteil.

Lawsonia (L.) intracellularis, an obligately intracellular bacterium, causes proliferative enteropathy (PE) in swine and, occasionally, in other animals. To determine the spread of the agent among German pig herds pooled fecal samples of five animals each of clinically normal Hessian pig herds collected between november 1998 and february 1999 as well as feces (n = 1684) from individual animals representing 648 herds, sent to our laboratory by veterinarians from all parts of Germany, were tested for L. intracellularis using the polymerase chain reaction (PCR). In addition, fecal samples from diarrhoic foals (n = 46), dogs (n = 57), cats (n = 50), calves (n = 37), hedge hogs (n = 9), seals (n = 8) and one giraffe were also studied. DNA was extracted from feces using high concentrations of chaotropic salt and diatomaceous earth. For PCR, primers flanking a 279 bp fragment of L. intracellularis DNA were used (JONES, G. F., WARD, G. E., MURTAUGH, M. P., LINN, G. (1993), J. Clin. Microbiol. 31, 2611-2615). Amplificates were separated by agarose gel electrophoresis and visualized under UV-light. L. intracellularis was found in 26 (12.8%) samples from 21 (30.0%) of the Hessian pig herds without symptoms of diarrhoea. In feces of pigs with diarrhoea (n = 1684) the agent was present in 431 (25.6%) samples originating from 224 (34.6%) herds. Of the other animal species studied, L. intracellularis was detected in feces of 4 (7.0%) dogs, 2 (5.4%) calves, 3 (33.3%) hedge hogs and in the sample of the giraffe. The remaining species were all tested negative.

Dog sera (n = 118) were tested for antibodies recognizing Borrelia (B.) burgdorferi sensu stricto strain B31 (ATCC 35210) antigens. In total, 18 of the dog sera gave positive results in a whole cell sonicate ELISA (WCS ELISA). These positive sera were further evaluated by immunoblot assay, utilizing a whole bacterial lysate as antigens. 94.4% (17 of 18) of the dog sera reacted with immunodominant antigens at 20-22 kDa (protein C, pC), 31 kDa (outer surface protein A, OspA), 34 kDa (outer surface protein B, OspB), 41 kDa (flagellin), 60 kDa ("common antigen"), and/or 100 kDa (presumably p100). Sera recognizing pC (20-22 kDa) and antigens94 kDa always detected the highest number of antigen bands, indicating the specificity of those antigens in serological diagnosis. The results clearly demonstrate that the WCS ELISA is a useful tool for testing sera of dogs for antibodies against B. burgdorferi. However, positive results should be confirmed by immunoblot, using WCS as antigen. According to the presented data, we recommend criteria for B. burgdorferi immunoblots using dog sera as follows: sera have to be considered as positive if they detect the 41 kDa flagellin, and two of the 5 immunodominant antigens, namely94 kDa (presumably p100), 60 kDa ("common antigen"), 34 kDa and 29-31 kDa (OspB and OspA, respectively) and 20-22 kDa (pC). If sera only recognize the 41 kDa flagellin, this result is equivocal, requiring testing a second serum sample 4 to 8 weeks later.

EHEC (enterohaemorrhagic E. coli) bacteria are new, only since 1982 recognized zoonotic pathogens. EHEC differ from E. coli intestinal commensales by the fact that they are lysogenic infected with bacteriophages, which carry the genetic information for the production of shigatoxins (Stx type 1 and/or 2). Due to the obligatory released Stx EHEC are classified also among the Shigatoxin producing E. coli (STEC). EHEC are capable of causing a Hemorrhagic Colitis and some sequelae of diseases such as the haemolytic uraemic syndrome. Due to their virulence factors they can be divided into typical and non-typical EHEC. Typical EHEC possess a pathogenicity island (Locus of Enterocyte Effacement) harboring genes, which apart from the characteristic necrotic activity of Stx enable the pathogens to closely attach to the epithelial cells of the intestinal mucosa and to destruct the microvilli. Additionally a so-called virulence plasmid codes for the production of a haemolysin, a peroxidase-katalase, an enterotoxin as well as a serine protease. EHEC are one of the world-wide most important causes of foodborne infections. Depending upon the country, most of the incidences in 1998 varied between 1 to 3 cases per 100,000 inhabitants. Since EHEC are only notifiable in a few countries, one must count however on substantially higher numbers. In Germany the estimated incidence is about 13 cases per 100,000 inhabitants. Since the first EHEC outbreaks were recognized in humans, studies investigating the prevalence of EHEC within animals were repeatedly performed. From the outset one assumed that cattle are a possible reservoir. Actually EHEC were isolated from fecal samples world-wide (typical and non-typical EHEC) from a large percentage of cattle (50%). Besides EHEC were isolated sporadically from fecal samples of other animals and healthy humans. The EHEC bacteria are shed by infected humans and animals, in particular by infected ruminants. They are spread over manure, slurry, sewage etc. Humans can get infected directly by contact with infected persons or animals or indirectly by contaminated food, water etc. The clinical outcome within humans appears as aqueous to bloody diarrhea. Beyond that approximately 5 to 10% of the patients develop the haemolytic uraemic syndrome. In contrast to humans, animals are mostly infected clinically inapparent. The therapy is based upon a symptomatic treatment. At present in man the control of EHEC infections concentrates on a particularly strict hand hygiene after the contact with infected humans and animals (above all ruminants). Since EHEC are heat sensitive, the prophylaxis by sufficient heating of risk food (raw milk, ground beef) is of special importance. In veterinary medicine above all EHEC infections must be controlled in ruminants, which are the primary reservoir. Due to the wide spread of EHEC in the ruminant population it is not realistic to demand an EHEC free cattle stock. Since EHEC are spread only via fecal excretion, at present it is most important to reduce the fecal shedding and to avoid fecal contamination of food of animal origin. In detail prophylactic hygienic measures concerning the farm management, the feeding hygiene, the food hygiene, the meat hygiene as well as the food hygiene are available.

Vom 26. bis zum 27. Mai 1998 fand das 15. Jenaer Symposium im Bundesinstitut fur gesundheitlichen Verbraucherschutz und Veterinarmedizin (BgVV) in Jena jachbereich BakterielleTierseuchen und Bekampfung von Zoonosen, statt.

An outbreak of edema disease (ED) was monitored in 80 piglets after weaning over a period of 4 weeks. The shedding of Shiga-like toxin-IIe) producing Escherichia coli strains, the serum bactericidal activity (SBA) against SLTEC-IIe, and the antibody response against SLT-IIe were investigated. The antibody response was monitored by utilizing a glutathione-S-transferase (GST) + SLT-IIe B/SUB fusion protein (FRANKE et al., in press) for immunoblot assays. E. coli-strain GO15III (0141:K85ac) was diagnosed as SLT-IIe-producing E. coli by polymerase chain reaction, DNA hybridization and cytotoxicity assays. Maximum excretion of GO15III appeared between days 8 and 15 after weaning. On day 1 after weaning no piglet shed GO15III, while the number increased on day 8 to 53 (66.2%) and on day 15 to 59 (73.8%) of the piglets. 4 week after weaning, GO15III was only isolated from 23 (28.8%) of the piglets. In parallel, serum bactericidal activity against GO15III increased significantly in the sera of 73 (91.2%) piglets, reaching a stable maximum from day 15 on. During the first two weeks after weaning, no piglet yielded detectable SLT-IIe-IgG. However, the number of SLT-IIe-IgG positive piglets increased steadily from day 15. On day 15, 5 (6.2%) piglets were positive in SLT-IIe immunoblot analysis and 29 days after weaning the number increased to 31 (38.8%). These data represent the first serological monitoring of a natural outbreak of edema disease in piglets after weaning by using a recombinant fusion protein (GST+SLT-IIe B/SUB). The recombinant protein proved to be a useful diagnostical tool for monitoring the specific antibody status of piglets.

A total of 56 liver specimens from rabbits with symptoms of rabbit haemorrhagic disease were tested for virus by electron microscopy (EM) and haemagglutination (HA). Both methods simultaneously gave positive or negative results in 28 or 22 cases, respectively. Divergent results were obtained in only 6 samples. Five of them were positive by EM but negative by HA and in one specimen with a HA-titer of 1:32 virus could not be detected.

From April 1990 through June 1991 clinical salmonellosis and asymptomatic faecal excretion of Salmonella spp. were seen in hospitalized horses at two veterinary hospitals. 76 Salmonella strains from hospitalized horses and 18 strains from horses without any clinical contact were characterized by serotyping and plasmid profile analysis. From April 1990 through January 1991 97.8% of the hospitalized horses were infected with strains of S. typhimurium var. Copenhagen, which were closely related according to their similar plasmid patterns. Other strains of S. typhimurium var. Copenhagen and serotype S. enteritidis were isolated not before February 1991. In the same period various plasmid profile types of S. typhimurium var. Copenhagen and strains of S. typhimurium, S. enteritidis and S. lexington were isolated from horses of the control group. Our results suggest that the high incidence of salmonellosis and latent salmonella infection in hospitalized horses was mainly due to the spread of one particular strain of S. typhimurium var. Copenhagen and that this strain was obviously acquired during hospitalisation.

Bacterial lysates of different bacterial strains (E. coli, B. bronchiseptica, P. haemolytica) were prepared by heating, acid- and alkaline-hydrolysis. Lysates were tested for their immunostimulating effect in bacterial infection models and with chromium 51 test demonstrating spontaneous (natural) cytotoxicity. Lysate production was standardized by protein- and Lps-determination. The alkaline-hydrolysis reduced toxicity of Lps and increased the content of soluble bacterial protein. Heating and acid-hydrolysis did not alter bacterial suspensions with respect to Lps-toxicity and protein-content. Mice infected with P. aeruginosa, P. multocida, E. coli and L. monocytogenes (5-10 LD50) had a significantly longer survival time after prophylactic immunostimulation with bacterial lysates than control animals. No protection was observed in immunostimulated mice infected with Erysipelothrix rhusiopathiae. In the Pseudomonas infection model, bacterial lysates prepared by alkaline-hydrolysis had a 10 times higher immunostimulating effect than lysates prepared by acid-hydrolysis or heating. Bacterial lysates stimulated spontaneous cytotoxicity of natural mouse peritoneal killer cells after intraperitoneal application. Whole bacterial lysates had a higher NK-activity as their corresponding purified lipopolysaccharide portion.

Kennel-specific oral E. coli vaccines were tested for efficacy and side effects at breeding and boarding kennels with severe diarrhoea problems. Oral vaccines contained heat inactivated E. coli bacteria from specific kennels and were given once daily for 14 days. Oral vaccines were administered directly orally to puppies while for adult dogs vaccines were mixed with daily food rations. In breeding kennels with 4 to 10 week old puppies suffering from non-fatal diarrhoea, the oral immunization led to a decrease in morbidity from 86.5% to 0%. In kennels with some cases of fatal diarrhoea the rate of morbidity decreased from 45% to 21% and the mortality rate from 25% to 10.3% after using the vaccination. By vaccinating adult dogs in boarding kennels the morbidity rate dropped from 83.5% to 6.5% and the mortality rate from 4.1% to 0.5%. The kennel specific oral E. coli vaccine was found to be free of side effects. No adverse effects were observed in either puppies or adult dogs.

Summary Efficacy of local and/or parenteral vaccination against calf salmonellosis with inactivated vaccines The S. typhimurium calf model was used to study immunization with inactivated vaccines against salmonellosis. Efficacy was demonstrated by comparison of post-infectious clinical reaction in vaccinated and non-vaccinated calves. The best immunization results were achieved with heat-inactivated bacteria or germ lysate. Formalin-inactivated bacteria were less effective. No immunity was observed after vaccination with phenol-water-extracted bacteria or enterotoxin preparation of S. typhimurium. Among the different application modes (nasal, dermal, oral, parenteral) combined oral and intramuscular vaccination proved to be most effective. The best immunization results could be achieved when calves were orally vaccinated once daily over 14 to 21 days in combination with an intramuscular booster immunization on the 10th day of the oral immunization period. Calves thus vaccinated were completely protected against 10 to 50 LD50S. typhimurium. Zusammenfassung Am Modell der S. typhimurium-Infektion des Kalbes wurde die Wirksamkeit von inaktivierten Salmonella-Impfstoffen untersucht. Der Wirksamkeitsnachweis erfolgte im Infektionsbelastungsversuch durch den Vergleich von Morbiditat und Mortalitat bei geimpften und nicht geimpften Tieren. Impfstoffe aus hitzeinaktivierten Erregern oder Keimlysaten ubertrafen in der Wirksamkeit Impfstoffe aus formalininaktivierten Keimen. Die orale Impfung mit einem Phenol-Wasser-Extrakt oder einer Enterotoxinpraparation von S. typhimurium blieb wirkungslos. Von verschiedenen Applikationsarten (oral, nasal, dermal, parenteral) hatte eine Kombination aus oraler und intramuskularer Impfung die beste Schutzwirkung. Taglich oral, uber einen Zeitraum von 14 bis 21 Tagen mit 1011hitzeinaktivierten S. typhimurium-Keimen immunisierte und einmal intramuskular geboosterte (10 Tage nach Beginn der oralen Impfung) Kalber waren gegen eine orale S. typhimurium-Infektion mit 10 bis 50 LD50 vollkommen geschutzt.

Ten splenectomized and non-splenectomized pigs were experimentally infected with E. suis bearing red blood cells in order to determine the antibody response. All animals were monitored for antibody titer by indirect hemagglutination over a period of 80-290 days postinfection. Latent E. suis infection only yielded a detectable antibody titer in one pig. Acutely infected pigs had a titer ranging up to 1:640. Maximum antibody response lasted only 2 months and dropped below the level of detection of our assay within 2 to 3 months. At this time, the clinical symptoms could reappear and antibodies were again detectable. However, no booster effect was observed with this second outbreak. We also determined the antibody frequency in 138 pigs from 16 herds in Southern Germany. Pigs from only 4 out of 6 clinically positive herds had antibody titer against E. suis. 20 out of 78 pigs of the clinical positive herds demonstrated a detectable E. suis antibody titer. In 10 herds that were asymptomatic and presumed uninfected all 80 pigs were serologically negative for E. suis.

Summary Comparative investigations on the distribution of plasmids in E. coli strains from dogs with diarrhoea, healthy dogs and their owners A total of 614 E. coli-strains taken from the faeces samples of healthy dogs, dogs with diarrhoea and their owners were examined for plasmid content. The plasmid patterns were shown by agarose electrophoresis. Around 20% of the E. coli strains from the faeces samples were free of plasmid. The remaining 80% of the strains had up to 13 plasmids displaying 11 different distribution patterns altogether. Typical plasmid patterns for E. coli strains from healthy dogs, dogs with diarrhoea and their owners could not be demonstrated. ETEC-strains were only isolated from sick dogs. These strains all contained plasmids. Hemolysin-forming E. coli strains appeared in three of the groups examined and either contained plasmids or proved plasmid-free, as with the mucoid-growing strains. On comparison of the E. coli strains from faeces samples of dogs and their owners, it was possible in the case of one family to isolate strains with identical plasmid patterns from the dog and the child. In 4 other families, individual plasmids of the E. coli strain from faeces samples of the dog matched those of the family members. Zusammenfassung Insgesamt 614 E. coli-Stamme aus Fazesproben von gesunden und durchfallkranken Hunden sowie von Hundebesitzern wurden auf ihren Plasmidgehalt untersucht. Die Darstellung der Plasmidmuster erfolgte mittels Agarosegelektrophorese. Rund 20% der E. coli-Stamme aus den Kotproben der Hunde und Besitzer waren plasmidfrei. Die ubrigen 80% der Stamme hatte bis zu 13 Plasmide, die ingesamt 11 verschiedene Verteilungsmuster aufwiesen. Typische Plasmidmuster fur E. coli-Stamme aus gesunden und durchfallkranken Hunden sowie Hundebesitzern konnten nicht nachgewiesen werden. ETEC-Stamme wurden nur aus kranken Hunden isoliert. Alle diese Stamme besasen Plasmide. Hamolysinbildende E. coli-Stamme traten in allen drei untersuchten Gruppen auf und waren sowohl plasmidfrei als auch plasmidhaltig, ebenso die mucoid wachsenden Stamme. Beim Vergleich der E. coli-Stamme aus Kotproben von Hunden und den Besitzern konnten in einer Familie beim Kind und beim Hund Stamme mit identischen Plasmidmustern isoliert werden. In 4 anderen Familien stimmten einzelne Plasmide der E. coli-Stamme aus Kotproben des Hundes mit denen der Familienmitglieder uberein.

The risk of a bacterial invasion by pseudomonas, e. coli, proteus, citrobacter and gaffkya via an open skin wound was investigated in combined injured mice (infliction of a skin wound and irradiations). Suspensions of different bacterial strains were applied on the wound and then the lethality was determined. Besides of apathogenic (citrobacter, proteus, gaffkya) and high pathogenic bacteria (e. coli and pseudomonas) an e. coli strain of the 0-Group 81 was found that seems to induce a decrease of lethality even after sublethal whole body irradiation.

The model "protective tetanic vaccination in mice" was used in order to try whether the paramunity inducer Pind-Avi may diminish or suspend radiation-induced immunosuppression. The radioprotective effect was ascertained from the different results of immunization in mice exposed to radiation after pretreatment with either Pind-Avi or placebo. The dose delivered amounted to 300 R. Tetanus toxoid was used for immunization. Effectiveness of the tetanic vaccination was analysed by means of load tests with 10 LD50 tetanus toxin, and of antibody determination with passive hemagglutination. Evidence was shown of significant radiation protection following subcutaneous application of Pind-Avi once a day during three days before of after exposure. Efficiency in mice treated with Pind-Avi amounted to 75% (pre-irradiation treatment) and to 43% (postirradiation treatment), whereas the efficiency in controls was zero in all cases. Thus, it was possible with Pind-Avi to suspend radiation-induced immunosuppression within a period of 3 days before or after radiation exposure, not only by means of prophylactic but also by therapeutic administration.

Infectious diarrhea in piglets can be caused by TGE, EVD and rota virus. The present report shows another viral agent (para-rota virus) to be present. In addition the report describes light and electronmicroscopic findings of the enteric mucosa caused by the agents mentioned. The degree and distribution of villous atrophy, one of the main histological findings, of infections with TGE, EVD, rota and para-rota virus is compared. A correlation of the results obtained with fluorescent antibody staining and ultrastructural diagnosis of agents was done. An application of the latter seems advisable in case of negative FA staining results.

Mandatory vaccination against smallpox was abolished on the account of smallpox-eradication proclaimed by the WHO and the postvaccinal complications detected after smallpox vaccination. At the same time vaccine banks with the vaccinia virus strain "Elstree" were organized. Should mass vaccinations with this vaccinia virus strain be carried out in a case of emergency, severe postvaccinal diseases and complications can arise in overaged and immunosuppressed vaccinees after primovaccination. Therefore attenuated vaccinia virus strains should be used for vaccine banks, which cannot be activated, or increase in virulence in impaired vaccinees after primovaccination. For these individuals the vaccinia virus strain "MVA", among other attenuated vaccinia strains, is recommended. The MVA virus strain can be applied parenterally without complications. From the scientific and field-relevant point of view it was tried to combine the vaccinia virus strain "MVA" with tetanus toxoid and to develop a combination vaccine "tetanus-smallpox". In immunization experiments using mice, piglets and monkeys, safety and efficacy of the vaccine were investigated. Efficacy was demonstrated by means of postvaccinal antibody determination and by the mouse protection test. Tetanus antitoxin was measured by ELISA and indirect hemagglutination test, antibody levels to vaccinia virus were investigated employing the neutralization test and hemagglutination inhibition test. No significant differences in potency could be demonstrated between the combination vaccine and the corresponding monovalent vaccines in mice, piglets and monkeys. The combination vaccine consisted of 12 Lf tetanus toxoid and 10 TCID50 vaccinia virus "MVA" preserved with gelatine and glucosamine. The double intramuscular immunization of monkeys stimulated average tetanus antitoxin titers of 1:310 and average vaccinia virus titers of 1:195 2 weeks p. revacc. Similar results were obtained in mice and piglets. Side reactions were not observed in mice and piglets. Except for occasional local reactions of short duration at the injection site of the monkeys, similarly no adverse reactions were observed after intramuscular vaccination with the combination vaccine.

The effect of rotavirus and enterotoxigenic Escherichia coli, administered in different sequences, on alkaline and acid phosphatase, leucinaminopeptidase, P-galactosidase, and succinicdehydrogenase of the intestinal mucosa of cesarian-derived, colostrum-deprived calves was investigated. Decrease in enzyme activity was most prominent in dual infections; it also occurred in parts of the small intestine in monoinfected animals. Increases in enzyme activity involved totally either one or all tissue compartments (crypt, basal villus area, villus tips). Increased activity was present in enteric mucosae that were either not affected or were only slightly affected by rotavirus or enterotoxigenic E. coli. We interpret the increase in enzyme activity as an adaptation of the enteric mucosa to maintain the absorptive function. An altered absorptive capacity of the enteric mucosa is one of the most important functional derangements leading to diarrhea. This can be studied in vivo by using tracer substances like D-xylo~e~~ or cellobiose mannit01.~~ In vitro, however, a large number of dif- ferent enzymes can be studied bi~chemically~,~ J~,~~~~~ or histo~hemically.~~,~~ Decreases in mucosal enzyme activities occur in neonatal diarrhea of calves.l~7~9J3~21~22,43 These histochemical results have been based on con- ventionally reared calves undergoing naturally ac- quired infections in the field.',22,43 However, in calves,27 enzyme histochemical studies have been reported on experimental enteric infections with various patho- gens. In the present study, five enzymes of primary im- portance for intestinal nutrient uptake were chosen.29 Enterocyte membrane bound enzymes examined were acid and alkaline phosphatase which are involved in sugar and lipid leucinaminopeptidase which is involved in peptide cleavage,'O and P-galac- tosidase which degrades lacto~e.~ The fifth enzyme, succinicdehydrogenase, is an intracytoplasmic enzyme that has been considered an indicator of intact cell metabolism. l3 A new technique combining conventional enzyme histochemistry and morph~metry~~ was used to eval- uate not only changes of the total enzyme activity at various locations in the small intestine, but also to give more information on enzyme activity in the main func- tional compartments of the mucosa (i.e., crypt, villus base, and villus tip). These relationships should give a better understanding of enzyme loss during infection and a possible regeneration during recovery.

The protective effect of a combined rotavirus/enterotoxigenic E. coli K99 vaccine against naturally occurring neonatal diarrhoea was evaluated in two dairy and two beef herds. Before vaccination, frequency of diarrhoea in these herds was 46% up to 60% with losses of 7% to 20%. After vaccination of pregnant cows and heifers mild diarrhoea occurred in 14% to 20% of their calves, one out of 144 calves died with enteric symptoms. In 13 out of 26 calves of nonvaccinated controls where diarrhoea was noted, one of them died. K99-positive enterotoxigenic E. coli were not detected in any of the faeces tested. Rotavirus was found in one of the beef herds.

Histopathologic and electron microscopic examination of intestines of three calves and two cats revealed attaching effacing bacteria characteristic of enteropathogenic Escherichia coli (EPEC) in ileum, cecum, and colon. The attaching effacing bacteria in one of the calves contained bacteriophages, and an E. coli isolate from that calf was shown to produce Shiga-like toxin. These findings contribute to emerging evidence that attaching effacing intestinal bacteria are globally distributed pathogens in a variety of host species and that bacteriophage-mediated production of Shiga-like toxin is related to the virulence of such bacteria.

There are adhesive and invasive species among enteropathogenic bacteria for animals. Most frequently isolated adhesive bacteria are enterotoxigenic E. coli (ETEC). Salmonella, T. hyodysenteriae, M. paratuberculosis, Y. enterocolitica, Cl. perfringens and C. jejuni bacteria represent the most important invasive germs. The main clinical finding connected with bacterial enteritis is diarrhoea resp. dysentery. Evaluation of feces consistency, time of appearance and accompanying symptoms (e.g. fever, colic) allow a clinical suspective diagnosis, however a definitive diagnosis must be based on the demonstration of the causative agent. In addition to known cultural, microscopic, biochemical and serological diagnostic methods, the demonstration of virulence factors (enterotoxin formation, adhesive pili) plays an important role in diagnosis of enteropathogenic bacteria.

Model experiments in mice were used for the investigation of oral immunization against tetanus. The efficacy of oral tetanus toxoid application was tested by subcutaneous challenge (10 LD50 tetanus toxin) and antitoxin determination by L+-method. Reasonable protection could only be achieved when toxoid was in intensive contact with the mucous membranes of the oral cavity, whereas immunization by way of the gastrointestinal tract failed completely. After a single oral vaccination with 200 Lf fluid tetanus toxoid onto mucous membranes of the oral cavity the efficiency index was 75. A 100% protection was obtained in challenge after 3 applications of 200 Lf toxoid. In comparison, a single subcutaneous dose of 2 Lf tetanus toxoid resulted in an efficiency index of 100 and a single intranasal immunization with 100 Lf in an efficiency index of 93. Development of immunity after oral immunization was much accelerated if compared with parenteral immunization. 20% of the orally treated animals were immune 5 days after vaccination. After the same period subcutaneously treated animals had failed to develop any protection at all. Antitoxin titers after one oral vaccination (200 Lf) were similar to the ones after one subcutaneous dose (2 Lf). A distinct increase in antitoxin titers could only be observed when revaccination was done subcutaneously. The results obtained with mice apply analogically to guinea pigs.

Model-experiments in mice were used for the investigation of oral immunization with tetanus-toxoid. The efficacy of oral toxoid application was tested by challenge and anti-toxin determination. The oral immunization with tetanus-toxoid induced only a sufficient protection, when toxoid was in intensive contact with the mucous membranes of the oral cavity. Local immunization via gastro-intestinal-tract failed. After a single vaccination with 200 Lf tetanus-toxoid onto mucous membranes of the oral cavity the efficiency index was 75. In comparison, a single subcutaneous application of 2 Lf tetanus-toxoid resulted in an efficiency index of 100 and a single intranasal immunization of 100 Lf in an efficiency index of 93. So the application of toxoid subcutaneously or onto nasal mucous membranes induced higher protection than oral application. Temporary development of immunity after immunization via the mucous membranes of the oral cavity was much better than parenteral immunization. 20 per cent of orally immunized animals were immune 5 days p. vacc. At the same time subcutaneously immunized animals failed to develop any protection. The efficiency rates obtained in challenge correlate with antitoxin content in the serum.

Under field conditions, the comparative effectiveness of oral, nasal and intramuscular vaccination of humans with tetanus toxoid was investigated. The effectiveness was tested by the determination of antitoxin titers with the L+-method. Oral booster vaccination produced only very low and irregular increases in antitoxin titer even at the highest dose of 6,000 Lf toxoid. In the most favourable case following triple oral immunization, a rise from 6.2 to 6.6 I.U. was achieved. Primary oral vaccination failed to stimulate measurable antitoxin production. After nasal booster vaccination with 1,000 Lf toxoid, a ten-fold rise in antitoxin titer was detectable. Following primary nasal vaccination, an average neutralization titer of 1 I.U. was achieved. The results of intramuscular and nasal booster vaccinations were similar although the antitoxin titers were significantly higher after primary intramuscular vaccination than after primary nasal vaccination. The average antitoxin titer amounted to 11.6 I.U. These results indicate that primary and booster vaccination against tetanus by the nasal route is a safe and effective procedure.

Ten splenectomized and non-splenectomized pigs were experimentally infected with E. suis bearing red blood cells in order to determine the antibody response. All animals were monitored for antibody titer by indirect hemagglutination over a period of 80-290 days postinfection. Latent E. suis infection only yielded a detectable antibody titer in one pig. Acutely infected pigs had a titer ranging up to 1:640. Maximum antibody response lasted only 2 months and dropped below the level of detection of our assay within 2 to 3 months. At this time, the clinical symptoms could reappear and antibodies were again detectable. However, no booster effect was observed with this second outbreak. We also determined the antibody frequency in 138 pigs from 16 herds in Southern Germany. Pigs from only 4 out of 6 clinically positive herds had antibody titer against E. suis. 20 out of 78 pigs of the clinical positive herds demonstrated a detectable E. suis antibody titer. In 10 herds that were asymptomatic and presumed uninfected all 80 pigs were serologically negative for E. suis.

Using the "tetanus-mouse" model the differences in immunity development between animals exposed to irradiation (300 R and 400 R) and animals not exposed to irradiation after nasal and subcutaneous vaccination was investigated. Immunization was carried out 1, 3, 6 and 10 days before and after exposure to irradiation. Efficacy of immunization was tested by challenge with 10 X LD50 tetanus-toxin and by antitoxin determination with L+-method. The degree of the immune response was dependent on 1. the irradiation dose, 2. the interval between active immunization and the ensuing or preceding irradiation, 3. the mode of vaccination (local or parenteral) and 4. the vaccination dose.

Hitherto the term "non-parenteral" (local) vaccination meant all those modes of vaccination where the vaccine is applied on the skin or the mucous membranes. The most common types of local immunization are oral and nasal vaccination. Using model studies with the mouse immunization per wound was investigated as an alternative mode of vaccination. Immunization was done with tetanus-toxoid, efficiency was tested by challenge with 10 LD50 tetanus toxin. For immunization per wound only slightly higher antigen doses are necessary compared with subcutaneous immunization. A single subcutaneous vaccination with 2 Lf tetanus-toxoid and a single application per wound of 10 Lf resulted in the same efficiency index of 100. Immunization per wound has to be carried out immediately after injury. It seems to be optimal within the first 3 to 5 hours. At a later time the same efficiency index can only be achieved by use of a higher antigen doses. Immunization per wound is possible with all kinds of wound, even with burns. Size of wound and wound-supply do not influence the efficacy of wound-immunization. Immunization per wound does not disturb wound healing. The experiments reported here showed, for the first time, that immunization per wound is as effective as parenteral immunization. The most important advantage of wound-immunization is the speed and ease with which it can be administered, a fact which predestines it for vaccination in emergency cases. In practice it will be useful to combine wound-vaccination with wound-sprays.