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One of the major goals of this study was to establish fast, reliable and sensitive assays for the quality control of immunomodulating phytopreparations and to determine whether pharmacological compounds or phytopreparations have effects on bovine immune cells. Flow cytometric methods were chosen because they are very sensitive in the detection of even subtle effects on cells. In this study, we addressed the question of whether these methods are useful in monitoring the effects of EquiMun and its compounds on bovine leucocytes in vitro. EquiMun is a fixed combination of Echinacea purpurea (Ec), Thuja occidentalis (Th) and elemental phosphorus (Ph) in different starting concentrations. Separated blood mononuclear cells (MNC) and polymorphonuclear cells (PMN, mainly neutrophils) were cultured for up to 44 h in vitro in the presence or absence of the tested substances. Whereas MNC were not affected by any of the compounds, EquiMun, Ec, Th and Ph significantly reduced the forward scatter (size) of cultured PMN without affecting their side scatter (granularity). The size effects were paralleled by a significantly enhanced viability of PMN after 20 h in culture. The observed effects were constant over wide concentration ranges and indicate a very similar reaction of leucocytes from individual cows. Whereas spontaneous generation of reactive oxygen species (ROS) by neutrophils was up-regulated by Ph and EquiMun, EquiMun down-regulated the phorbol ester-stimulated ROS production. However, ROS generation by neutrophils displayed a large inter-individual variation with less apparent, down-regulatory effects of EquiMun. The ability of PMN to kill target cells via antibody-independent cellular cytotoxicity showed small inter-individual variations and was enhanced by Ec and Th but not by Ph and EquiMun, probably due to dose-dependent effects. In summary, the flow cytometric characterization of cellular viability and shape changes of neutrophils seem to be a suitable and reliable approach for the quality test of immunomodulating phytomedicines based on bioassays.

Summary Influence of oral dosing with vitamin E on parameters of defence against infection in pigs at the early fattening stage Alpha‐tocopherol‐acetate was supplemented to the food of pigs during first fattening‐period in doses of zero, 150 and 300 mg/kg of food. Parameters of resistance to infection were tested in 180 pigs under test on five different times during 10 weeks. Pigs fed on a diet supplemented with 150 mg Vit. E showed higher activity of lysozym and a higher rate of lysis of sacc. cer. as both other groups. Differential blood count done in 30 pigs showed relative and absolute higher concentration of granulocytes in blood of pigs fed on 150 mg Vit. E, whereas the serum‐IgG‐level was significantly decreased by this 150 mg‐dose. The hypothesis of a non‐linear stimulation of unspecific defense‐mechanisms and the retardation of the age‐dependent physiological decrease of granulocytes by the 150 mg‐supplementation of Vit. E is discussed. Zusammenfassung Die Wirksamkeit von alpha‐Tocopherol‐Azetat‐Supplementierungen in Dosierungen von 0, 150 und 300 mg pro kg Alleinfutter auf verschiedene Parameter der Infektionsabwehr wurde in einem Blindversuch an insgesamt 180 Schweinen während der Anfangsmast unter praxisüblichen Keimbelastungen geprüft. Die Abwehrparameter wurden zu fünf unterschiedlichen Zeitpunkten bestimmt. Die 150‐mg‐Dosierung des Vitamin E bewirkte eine höhere Lysozymaktivität und eine höhere Hefezytolyserate als die beiden Vergleichsdosierungen. Das bei 30 Tieren untersuchte Differential‐blutbild zeigte eine relativ und absolut höhere Granulozytenkonzentration im Blut der 150‐mg‐Gruppe. Dagegen wird der IgG‐Gehalt des Serums durch die 150‐mg‐Dosierung signifikant vermindert. Die Hypothese einer nicht‐linearen Stimulation unspezifischer Abwehrsysteme und die Abpufferung eines entwicklungsphysiologisch bedingten Rückganges der Granulozyten durch die 150‐mg‐Dosis des Vitamin E werden diskutiert.

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