Flow Cytometric Testing of Immunological Effects of a Phytomedicinal Combination (EquiMun) and its Compounds on Bovine Leucocytes

Abstract

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.

Dietary supplementation with Echinacea and development of immunity to challenge infection with coccidia

“…There are numerous recent reports indicating the immunostimulating or modulating effects of various preparations and extracts (Stimple et al 1984;Burger et al 1997;See et al 1997;Sun et al 1999;Currier and Miller 2001;Goel et al 2002). There are a few reports of the veterinary application of Echinacea therapy, for example, in horses (O'Neill et al 2002 ), swine (Stahl et al 1990) and cattle (Schuberth et al 2002) . Nothing has been reported on its use in poultry.…”

Effect of supplementation of Satureja essential oils in drinking water on immune performance of broiler chickens reared under heat stress

“…In some studies carried out in animal fields, the influence of medical plants on improvement of immune system had been reported (Rivera et al, 2003;Schuberth et al, 2002). Furthermore, Rahimi et al (2011) reported that dietary thyme extract (0.1%) soluble in water did not affect immune system, but relative weight of bursa fabricius was significantly affected by garlic group and antibody response to SRBC was higher in coneflower group when compared with control group (P<0.05).…”

Efficiency of savory (Satureja Khuzestanica Jamzad) essential oil on performance, carcass traits, some blood parameters and immune function of Male Ross 308 heat stressed broiler chicks

“…Mohammad et al (2013) reported that, inclusion of savory essential oil for 21 days (21-42 days) in diet of heat stressed broilers had no significant effect on lymphoid organs (spleen, thymus, bursa of Fabricius) weight and our findings confirm it. Besides, in some studies performed in animal fields, the influence of medical plants on improvement of relative weights of different organs had been reported (Rivera et al, 2003;Schuberth et al, 2002). Additionally (Rahimi et al, 2011) reported that the dietary thyme extract (0.1%) soluble in water affected relative weight of bursa of fabricius of broilers significantly.…”