Arch.Geflgelk., 73 (4). S. 227236, 2009, ISSN 0003-9098.

Verlag Eugen Ulmer, Stuttgart

Echinacea: A potential feed and water additive in poultry and swine production
Echinacea: Ein potentieller Futter- und Trinkwasserzusatz in der Geflgel- und Schweinerzeugung
Z. Nasir and M.A. Grashorn
Manuskript eingegangen am 5. August 2008, angenommen am 17. Oktober 2008

Introduction
The use of growth promoting antibiotics in animal feeds is prohibited in European Union (EU) since 2006, due to the risks associated with their use and development of resistant strains of bacteria. Now, as a result of this ban and increasing demand of organic animal products, alternative substances are required. These alternatives should be beneficial in maintaining health, improving immunity and performance of farm animals without any residual effects on consumer health and environment. A number of experiments conducted to find alternative substances revealed that no single alternative exists with the effects comparable to antibiotics (NASIR and GRASHORN, 2006). Plant extracts contain numerous substances having pharmacological activities. Especially for essential oils a wide range of effects have been proven, including inhibitory action on pathogens, effects on physiopathologies (e.g. anti-inflammatory, anti-diarrhoea properties) and activity in different body systems e.g. endocrine and immune system (RECOQUILLAY, 2006). Echinacea (E) is a North American indigenous plant (family Asteraceae), which has been traditionally used as a cure all for different ailments. In recent years, it has become one of the most popular herbal products in North America and Europe as an immune promoter and immune stimulator (GOEL et al., 2002). Echinacea extracts and preparations have been used traditionally for individuals suffering from sore-throats, coughs, and various respiratory symptoms that could be due to bacterial infections (SHARMA et al., 2008), with the proposed mechanism of action relating to its reported ability to stimulate the immune system (BARRET, 2003). E. purpurea (purple coneflower) is the best known of the dozen or so species of the genus Echinacea and is cultivated widely in North America, Asia, and Europe (especially in Germany) for its beauty as well as for its reported medicinal properties and ability to stimulate immune system (BARRET, 2003). Other important species are E. pallida (pale purple coneflower) and E. angustifolia (narrow-leaved purple coneflower, black Samson Echinacea).

Chemical composition of Echinacea

Chemical composition and biomass of Echinacea varies not only among different varieties, but environmental conditions and agronomical practices also have effect on them. Table 1 represents the biomass partitioning of E. purpurea and micro mineral levels in its leaves. A variety of pharmacologically active substances are present in different plant parts, juices, extracts, and preparations of Echinacea (Table 2). The level of these constituents varies in different plant parts as well as among varieties, and helps in differentiating between preparations made from various plant parts and different species. Major pharmacologically active compounds present in Echinacea and its preparations are alkamides, glycoproteins, polysaccharides, phenolic compounds and cinnamic acids (LIU Y-C et al., 2007; HARBORNE and WILLIAMS, 2004).

Alkamides
Alkamides (fatty acid amides) are the major lipophilic, bioactive phytochemicals found abundantly in certain species of Echinacea, especially in high concentrations in the roots of E. purpurea, E. pallida and E. angustifolia (LALONE et al., 2007). Alkamides are characteristic rhizome components of E. angustifolia and the rhizomes and aerial parts of E. purpurea. Alkamides are the major constituents of the ethanol/water extract of Echinacea and are widely used for therapeutic purpose. They are likely to cross the intestinal barrier (MATHIAS et al., 2004) as they are found in the blood stream of patients who ingested Echinacea extracts (DIETZ et al., 2001).

Glycoproteins
Three glycoproteins have been isolated from E. angustifolia and E. purpurea roots (BEUSCHER et al., 1987). Glycoproteins have been implicated in the immunostimulatory activity of Echinacea extracts by inducing cytokine production and by their mitogenic activity (BAUER, 1993; 1994).

Polysaccharides
Two polysaccharides with immunostimulatory properties have been isolated from the aerial parts of E. purpurea. However, from tissue culture of E. purpurea three homogeneous polysaccharides, two neutral fucogalactoxyloglucans and an acidic arabinogalactan were isolated (WAGNER et al., 1988). Fucogalactoxyloglucans enhanced phagocytosis both in vitro and in vivo, while arabinogalactan spe-

Dept. of Poultry Science (470 c), University of Hohenheim, Stuttgart, Germany

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Table 1. Characteristics of Echinacea purpurea plant and micro minerals in Echinacea purpurea leaves Charakteristika der Pflanze Echinacea purpurea und Gehalte an Mikronhrstoffen in den Blttern von Echinacea purpurea
Biomass DM Percentage of DM Roots 1535

300400 g/plant 2526%

Flowers 1535

Shoots and stem 2025

Leaves 2530

Micro minerals in Echinacea purpurea leaves (mg/kg)

B Cu Zn Mn Fe P

329

21

23

81

1,388

3.3

K 396

(BONOMELLI et al., 2005)

Table 2. Contents of alkamides, chicoric acid and total phenolics in various plant parts and corresponding extracts of E. purpurea Gehalte an Alkamiden, Chicoresure und Gesamt-Phenolen in verschiedenen Pflanzenteilen und extrakten von E. purpurea
Plant part Alkamide g/g DM Chicoric acid mg/g DM 41.3 8.57 7.63 8.17 10.1 Alkamide mg/ml extract Total phenolics mg/ml extract 1.62 0.30 0.37 0.42 0.48 Chicoric acid mg/ml extract 1.65 0.34 0.31 0.33 0.40

Leaves Stem Root stock Main root Side root (THYGESEN et al., 2007)

2.2 4.2 0.5

0.09 0.17 0.02

Table 3. Pharmacological activity of Echinacea Pharmakologische Wirkung von Echinacea

Activity Anti-inflammatory Preparation/part used Alkamides derived from E. angustifolia Different Echinacea preparations and extracts Commercial E. purpurea preparations (root or/and aerial) Commercial E. angustifolia root preparations Polysaccharides from E. purpurea Complete herb and roots of E. purpurea Reference HINZ et al., 2007 BIRT et al., 2008; WOELKERT and BAUER, 2007; LALONE et al., 2007 SHARMA et al., 2008

Antibacterial (Human pathogenic bacteria) Macrophage activation (phagocytosis) PMN granulocytes modulation NK cells stimulation

E. purpurea root E purpurea extracts

BARRET, 2003 JURKSTEINE et al., 2004; ZHAI et al., 2007; GROOM et al., 2007; SULLIVAN et al., 2008.. MILLER and YU, 2004 CURRIER and MILLER, 2001; BROUSSEAU and MILLER, 2005; WOELKERT and BAUER, 2007

cifically stimulated macrophages to excrete the tumor necrosis factor (TNF) (WAGNER et al., 1988).

roots of both E. purpurea and E. angustifolia provides a useful means of differentiating root preparations of E. pallida from other two species.

Hydrocarbons
These are characteristic root constituents of E. pallida, where some 11 derivatives, mainly ketoalkenes and ketoalkynes (polyacetylenes) have been identified (HARBORNE and WILLIAMS, 2004). The only hydrocarbon reported from roots of E. purpurea is an ester (dodeca-2, 4-dien-1-yl isovalerate) which is also present in roots of E. angustifolia (HEINZER et al., 1988). The absence of polyacetylenes from

Essential Oils
The roots of E. purpurea have been reported to contain up to 0.2% essential oils (BAUER, 1999). The major components were found to be caryophyllene (2.1%), humulene (0.6%), and caryophyllene epoxide (1.3%) (BECKER, 1982). The flowering aerial contain less than 0.1% essential oil. The constituents identified include borneol, bornyl aceArch.Geflgelk. 4/2009

Nasir and Grashorn: Echinacea as feed and water additive in livestock production
tate, germacrene D, and caryophyllene and its epoxide, which were present also in aerial parts of E. pallida and E. angustifolia (BAUER, 1999). The main essential oils in all the plant tissues, irrespective of species, were camphene, -pinene, and limonene, together with other volatiles such as acetaldehyde, dimethyl sulphide, and hexanal (HARBORNE and WILLIAMS, 2004).

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Phenolic compounds
Caffeoyl-quinic and caffeoyl tartaric acid esters are characteristic phenolic constituents of E. purpurea, E. angustifolia and E. pallida. Chicoric acid is the major component in all three plant parts of E. pallida, while echinacoside is present in an equally high concentration in the roots but only in moderate amounts and trace amounts in flowers and leaves (HARBORNE and WILLIAMS, 2004). Total phenolic contents were highest (23.2 mg/g) in roots of E. purpurea as compared to roots of E. angustifolia (10.5 mg/g) and E. pallida (17.8 mg/g) (FEDERICA et al., 2003). Some researchers have reported that chicoric acid is more abundant in the flowers, with much lower contents in leaves and stem (BAUER et al., 1988). The leaves have been shown to additionally contain chichoric acid methyl ester, 2-caffeoyl-3-feruloyltartaric acid, 2, 3-diferuloyltartaric acid, 2-feruloyltartaric acid and 2-caffeoyl-3-p-coumaroyltartaric acid (BECKER and HSIEH, 1985; SOICKE et al., 1988). LIU et al. (2007) reported that the contents of caffeic acid derivatives in E. purpurea reach its highest in the middle stage of full blossoming and were higher in fresh raw material than in dried raw material. They observed that contents of chicoric acid vary from 0.1% (in dried roots) to 3.3% (in fresh flower buds), caftaric acid vary from 0.1% (in dried roots) to 0.72% (in fresh flower buds), and total phenolic compounds vary from 1.91% (in dried aerial parts) to 6.47% (in fresh flower buds).

Pharmacological properties of Echinacea

Echinacea is very popular for its effects on the immune system (HINZ et al., 2007). Results of some experiments studying pharmacological properties of Echinacea are summarized in Table 3. It has demonstrated its properties by stimulation of various immune cells such as macrophages, other monocytes, and natural killer (NK) cells during in vitro (BAUER, 1999; BURGER et al., 1997; RININGER et al., 2000; GROOM et al., 2007; SULLIVAN et al., 2008) as well as in-vivo studies (CURRIER and MILLER, 2001; 2002). Among the many pharmacological effects reported, modulations of macrophages, PMN (polymarphonuclear leukocytes) immune cells and effects on cytokines/chemokines expression in human cells have been demonstrated most convincingly (WOELKERT and BAUER, 2007). E. angustifolia (root extract) have been reported to enhance immune function by increasing antigen-specific immunoglobulin production (REHMAN et al., 1999). HINZ et al. (2007) found that alkamides derived from E. angustifolia roots may contribute to the pharmacological action of the herbal extract by inhibiting COX-2 activity at sites of inflammation. Echinacea extracts may be able to modulate inflammation through their inhibitory activity on PGE2 production and alkamides are possible key constituents in the observed anti-inflammatory properties, most likely acting additively or synergistically with other constituents (LALONE et al., 2007). Various extracts of E. purpurea showed different activities against human pathogenic bacteria (SHARMA et al., 2008). Analysis of polar fractions of E. purpurea extracts showed the presence of antiviral
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activity, with evidence suggesting that polyphenolic compounds other than the known HIV inhibitor, chicoric acid may be involved (BIRT et al., 2008). E. purpurea preparations have been reported to activate human macrophages (MSE, 1983) and enhance cytokine production (BAUER, 1999; BURGER et al., 1997; RININGER et al., 2000). Solutions made from herb and root E. purpurea powders produced definite reproducible macrophage activity, anti-inflammatory and antioxidant properties (RININGER et al., 2000, ZHAI et al., 2007). Increased activity of PMN granulocytes has been reported from a number of animal experiments (MILLER and YU, 2004). Mice fed 0.45 mg/day of E. purpurea root extract for 12 weeks showed significantly higher numbers of NK cells and monocytes, but no granulocytes, lymphocytes or their precursors (SUN et al., 2001). E. purpurea extract has been reported to increase both antibody-dependent and innate NK mediated activities against herpes virus infections in ex vivo cells from both normal and immune-depressed individuals (SEE et al., 1997). Increased white cells counts in peripheral blood have been noted following intramuscular or intravenous injection of E. purpurea extracts, but not following oral dosing (LORENZ et al., 1972). E. purpurea preparations from roots activate the cellular immunity and stimulate phagocytosis of neutrophils in vitro, in vivo and after rinsing of mouth cavity (JURKSTIENE et al., 2004). Polysaccharides derived from E. purpurea are reported to enhance macrophage activity in mouse, rat and human (BARRET, 2003). Daily dietary administration of E. purpurea root extract to normal mice for as little as 1 week resulted in significant elevation of natural-killer (NK) cells; and showed profoundly positive effects in Leukemic mice, when administered daily for 50 days from the onset of Leukaemia (CURRIER and MILLER, 2001). TURNER et al. (2005) studied the effect of chemically defined extracts from E. angustifolia roots (equivalent of 300 mg root) on rhinovirus infection in 437 volunteers, and found that extracts of E. angustifolia root, either alone or in combination, did not have clinically significant effect on infection with a rhinovirus or on the clinical illness that results from it. However, a meta-analysis evaluating the effect of Echinacea on the incidence and duration of common cold showed that Echinacea decreased the odds of developing the cold by 58% and duration of cold by 1.4 days (SHAH et al., 2007). Dietary supplementation of E. purpurea daily (2 mg/mouse) from puberty (7 week) until just beyond 13 months of age (late middle age in mice) improved survivability of mice by significantly elevating levels of NK cells in their bone marrow production site, as well as in the major organ to which they traffic and function, i.e. the spleen. Thus, it appears that regular intake of E. purpurea (root extract) may indeed be beneficial and prophylactic (BROUSSEAU and MILLER, 2005). Basic research on Echinacea as a medicinal botanical identifies its major activity as a stimulant to the phagocytic potential of polymarphonuclear cells (WAGNER et al., 1988), which is a well established measurement of immune function (ATHLIN et al., 1991). Other immunologically relevant activities of Echinacea plants include inhibition of cyclooxygenase enzyme and 5-lipoxygenase enzyme (two key enzymes involved in the inflammatory response) (MLLER-JACKIC et al., 1994) and a stimulatory effect on the secretion of various cytokines, including IL-1, IL-6, and TNF- (LTIG et al., 1989; ROSLER et al., 1991, STEINMLLER et al., 1993; SULLIVAN et al., 2008). Echinacoside present in Echinacea exhibited a stronger protection against free radical-induced native collagen degradation than other caffeic acid derivatives, such as chichoric acid, caffeic acid, and chlorogenic acid (FACINO et al., 1995).

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preparations made from E. purpurea, which can be administered through drinking water as well as through feed, however presence of different amount of active ingredients in different preparations make the decision difficult. The dosage of Echinacea preparations for livestock species is not available, so due to lack of data, researchers adjusted dose level on the basis of human medical recommendations (0.25ml/kg BW0.75). The following overview provides some species specific information on the pharmacology, toxicity, and clinical application of Echinacea in various livestock species, including cattle, horse, poultry and swine.

Taken together, these data suggest that E. purpurea may have potential in stimulating the immune system response of humans, laboratory animals as well as livestock under normal and stressful husbandry conditions. Various extracts and purified preparations made from different plant parts of E. purpurea can be used to maintain health and improvement of immunity in animals. However, there is little consensus by which species, plant part, or extraction method the greatest immune-stimulating properties are achieved. Extracts soluble in water, ethanol and chloroform have all demonstrated phagocyte-stimulating activity (BARRET et al., 2003, BAUER, 1999).

Cattle Mode and level of administration

Echinacea is either used as dried whole plant; as plant parts; as plant extracts in ethanol, water or chloroform; as plant juices fermented or in alcohol; as dried plant products in powdered form with food/feed or in the form of tea; as plant parts alone or in combination with other plant parts or as a mixture with other species of Echinacea and even other herbal plant species (MILLER and YU, 2004). According to BONE (2004) different Echinacea products and preparations available in the market are: 1. The stabilized juice of E. purpurea tops (often sold under the trade name of Echinacin) 2. Fresh or dried whole plants, aerial preparations, root preparations of E. purpurea, E. angustifolia or E. pallida. 3. Mixture of any of above. Preparations of the above are applied in various dosage forms including liquids (in ethanol-water mixture or other), capsules, and dried extracts (tablets or capsules). Some preparations are administered by intramuscular injection in some countries, especially in Europe (BONE, 2004). Dosage and duration of application also vary from preparation to preparation and depends upon availability of pharmacologically active substances. No dose level or duration of Echinacea has been universally standardized. However German Commission E recommends E. purpurea aerial parts expressed juice, but not E. purpurea root; conversely, it recommends E. pallida alcohol root extracts, neither the roots nor aerial portions of E. angustifolia are recommended (BGA COMMISSION E, 1989). According to BGA COMMISSION E (1989) Echinacea should be prophylactically used daily for several weeks. However, JURCIC et al. (1989) observed that phagocytosis rate of granulocytes in human blood was increased during the first 5 days of application and depressed afterwards as a result of over-stimulation. Similarly, some beneficial effects of short-time (25 days), repeated application of E. purpurea juices have been reported by BHMER et al. (2008); NASIR and GRASHORN (2007a) in poultry and by KUHN et al. (2005) in swine production. However, continuous application of E. purpurea in poultry (ROTH-MAIER et al. (2005) and swine (MASS et al., 2005) did not show significant effects. Therefore, it seems that beneficial effects of application of Echinacea preparations on animal health and performance can be obtained by repeated short-time (35 days) application followed by application free days. SCHUBERTH et al. (2002) studied the effect of an E. purpurea tincture (containing 70% ethanolic extract of E. purpurea), Thuja occidentalis, and elemental phosphorus on bovine leucocytes. Bovine mononuclear cells (MNC) and polymarphonuclear (predominantly neutrophils) cells were isolated from cows and cultured for up to 44 hours in the presence and absence of an extract of the product and its individual components. Flow cytometry results showed no significant effect of individual components, which included Thuja occidentalis, E. purpurea, and elemental phosphorus, on MNC. However, E. purpurea was able to enhance the ability of the PMN to kill target cells via antibody-independent cytotoxicity. GILL et al. (2002) studied the effect of Echinacea supplementation on immune function in transitional calves. Ten weaned calves were assigned to treatments of 0 or 2.5 g/day of Echinacea supplementation in ration for 17 days. The monocytes percentage decreased (P = 0.0253) in treated calves over time and varied between breeds. B cell percentage also decreased (P = 0.0343) with treatment as well as with application time. CD4+ T cell levels significantly increased (P = 0.0253) with Echinacea supplementation and CD2+ T cells and white blood cells percentage tended to be higher in treated calves. Neutrophil contents, packed cell volume, serum protein concentration and body weight were not affected.

Horses
Along with a variety of herbs, Echinacea is also used as immune booster to compliment a healthy immune system of horses. According to WILLIAMS and LAMPRECHT (2007), best way to use Echinacea is to supplement at the first signs of illness or infection. If administered too late in the cycle the herb will be less effective. ONEIL et al. (2002) offered a standardized aqueous extract of E. angustifolia (prepared from powdered root that was standardized to 4% echinacoside) or an inactive placebo to eight healthy horses for a period of 42 days. Blood samples taken every 7th day were subjected to a complete haematology and biochemistry screen, and a phagocytic function test. Echinacea extract increased the production of lymphocytes and decreased the level of circulating neutrophils in the blood presumably by increasing membrane permeability and migration into tissues. Echinacea administration increased the number of red blood cells and haemoglobin, showing its effect on the oxygen transport mechanism.

Poultry Use of Echinacea in Livestock

Presence of a variety of pharmacologically active substances in E. purpurea and prove of their activity in vivo as well as in vitro experiments suggest that it may be beneficial in maintaining health and improving performance and immunity in livestock. There are a number of products and Research on application of Echinacea preparations in poultry is limited; however it can be used as a guiding line for future studies. NASIR and GRASHORN (2007a) studied the effects of application of two different E. purpurea juice preparations by drinking water to broilers (Ross 308) on performance and blood parameters for stress, protein and
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lipid metabolism and immune status. E. purpurea fermented juice (EFJ) and E. purpurea juice on alcohol (EOJ) basis were applied through drinking water at levels of 0.25 ml/kg BW0.75 and 0.50 ml/kg BW0.75. The dosage of the Echinacea juice for hens was adjusted on the basis of human medical recommendations (0.25 ml/kg BW0.75) due to lack of data for poultry. (Metabolic body weight is used for calculation of application level as it represent the weight of those tissues in body that metabolize most of the nutrients that are absorbed in the body; and to adjust the differences among the animals (human and poultry)). Duration of E. purpurea juices application was three days followed by treatment free days (8, 9 or 12). The results showed that E. purpurea juice supplemented groups performed better as compared to non supplemented groups. There was no significant difference in level of serum total protein, albumin and globulin between 0.25 ml/kg BW0.75 and 0.50 ml/kg BW0.75 treatments. Levels of ALT, -GT, alkyl phosphate, creatine kinase, LDH and serum total cholesterol were non significantly different. Blood picture also showed no significant treatment effect on number of leucocytes, erythrocytes, haemoglobin contents and hematocrit percentage. In another experiment, comparing the effects of E. purpurea fermented juice and E. purpurea juice on alcohol basis, applied through drinking water at the rate of 0.25 ml/kg BW0.75 and 0.50 ml/kg BW0.75 against a negative control, better feed conversion ratio was observed in E. purpurea treated groups as compared to control. Serum total protein level was non significantly higher and serum albumin concentration was lower than control group. The application of E. purpurea fermented juice at the rate of 0.25 ml/kg BW0.75 resulted in non significant improvement of feed conversion ratio and performance (NASIR and GRASHORN, 2007b, 2008). BHMER et al. (2008) studied the effect of intermittent application of E. purpurea juices in feed on layer performance and immune status. They found significantly (p < 0.05) increased number of lymphocytes and total leucocytes, but reduced phagocytic activity in groups receiving ethanolic E. purpurea juice for 5 days. Treatment of EFJ juice for 2 days (12 days interval) resulted in highest antibody titers as compared to EOJ. ROTH-MAIER et al. (2005) studied the effect of dietary supplementation of dried aerial parts of E. purpurea as feed additive in the diets of broilers and layers on feed intake, feed conversion efficiency and growth/egg performance. The aerial parts of E. purpurea plants were pressed to cobs and stored. These cobs were added to experimental diets of broilers and layers after grinding. Supplementation of 0, 0.6, 1.2, 1.8, 2.4, 3.0, 3.6, 4.2, or 4.8% E. purpurea cobs for a continuous period of 5 weeks to broiler diets showed no significant effect on recorded parameters. In another experiment with broilers treated with 0, 2.4% E. purpurea cobs or 10 mg/kg feed Flavomycin, E. purpurea supplemented groups showed lower feed intake and body weight gain compared with control and Flavomycin treated group. Supplementation of 0 or 1.8% E. purpurea cobs to layer pullets from 021 weeks and 0 or 4.8% E. purpurea from 21st week, showed no effect on body weight gain at week 4, 8, and 20, while feed intake of E. purpurea group was depressed in week 58 by 11%. In the egg production period, no difference between treatment groups was observed. The authors concluded that continuous supplementation of E. purpurea cobs is not beneficial for broilers and layers (ROTH-MAIER et al. 2005). However, Echinacea dietary supplements proved beneficial adjuvants for live anticoccidial vaccines (ALLEN, 2003). Effects of E. purpurea supplementation on weight gain before challenge, and weight gains, lesion scores and plasma levArch.Geflgelk. 4/2009

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els of carotenoids and NO2- and NO3- (which are indicators of macrophage activation and phagocytic activity) following challenge with multiple coccidia species (Eimeria acervulina, Eimeria tenella, Eimeria maxima and Eimeria necatrix) were determined. The results showed that, combined live vaccination and feed supplementation with 0.1% or 0.5% dried, ground root preparations of E. purpurea during the first two weeks of life provided significant weight gain advantage compared to live vaccination alone. This advantage persisted through two weeks of E. purpurea withdrawal and subsequently challenges infection. E. purpurea supplementation also lowered significantly total lesion scores but didnt significantly modify the effects of vaccination and challenge on plasma carotenoids or NO2- + NO3. The author suggested that E. purpurea has potential for use as adjuvant for live vaccines when provided as a dietary supplement concurrent with the application of a live vaccine. E. purpurea can also be used to provide protective immuno-stimulation in the presence of natural populations of coccidian in litter (ALLEN, 2003). SCHRANNER et al. (1989) studied the effect of the complex drug (Influx) and E. angustifolia extract in humoral immune response of intact and immunodeficient chickens. The preparations were administered in two oral doses, after which the complete immunoglobulin concentration and the antibody production were determined. The administration of the complex drug to normal Leghorn chicken induced a rise in the serum immunoglobulin concentration, as well as increase in the three classes of antibody. In immunodeficient chickens, the complex drug caused a slight production of IgG. GARDZIELEWSKA et al. (2003) studied the effect of plant supplementation feeding on fresh and frozen storage of broiler chicken meat. They supplemented 1% dried E. purpurea herb, 0.3% crushed raw garlic in feed. Another group received twice a week raw ginger water extract (5.5 g/cm3) to drink. Chemical composition of breast muscles were analysed freshly and after 4-month storage at 18C. Supplementation with E. purpurea or garlic did not show significant influence on dry matter, crude protein, or raw fat content in the breast muscles. E. purpurea supplemented groups showed numerically higher levels of crude protein, better pH value (5.5) and better, darker colour as well as better juiciness. Dried E. purpurea herb supplemented (560 mg/kg) in broiler grower diets from 22 to 42 days of age maintained flavour of meat after storage (KORELESKI and SWIATKIEWICZ, 2007). The results of the studies in poultry showed that efficacy of E. purpurea depend on its preparation (level of active substances) and on its mode of administration. Dried E. purpurea cobs supplementation in feed showed no significant effect on performance as well as immune status of broilers and layers (ROTH-MAIER et al. 2005), but dietary supplements of E. purpurea seem to be useful adjuvants for live anticoccidial vaccines (ALLEN, 2003) and improved performance when E. purpurea juices are administered through drinking water (NASIR and GRASHORN, 2007a, 2007b, 2008) and maintained meat quality during storage (KORELESKI and SWIATKIEWICZ, 2007). There is lack of research on using other application methods and E. purpurea preparations in poultry. Supplementation of E. purpurea extracts (ethanol or water) or liquid preparations (juices) can be more beneficial as compared to dried products, as they contain higher amounts of active substances which can be absorbed easily.

Swine
Preparations made from E. purpurea have been reported to improve swine health, performance, and meat quality.

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104 tissue culture infectious dose50/ml. Among PRRSV challenged pigs, dietary E. purpurea did not affect (P > 0.10) the rate or level of ELISA-detectable antibody response from day 7 to 42 or the level and duration of PRRSV in serum. For PRRSV- unchallenged animals receiving diets supplemented with E. purpurea at 2 or 4%, no differences (P > 0.10) were observed in average daily gain, average daily feed intake or gain: feed ratio. HOLDEN and MCKEAN (2002) conducted a series of experiments to compare the health and performance of weanling pigs fed varying levels of Echinacea with those receiving a subtherapeutic level of a common antibiotic (Mecadox). At the tested levels (0.01, 0.25, 0.5, 2.0 and 3.0%) in feed, no statistical advantage in weight gain or feed conversion was observed when compared with the diet containing 45 ppm of Mecadox or with negative control containing no antimicrobial or botanical inclusion. Echinacea treated pigs exhibited a light, but not objectionable off-flavour of their meat when compared to control. They observed that in weeks 0 to 3 and 0 to 4 the higher levels of Echinacea (0.5% and 2.0%) were significantly more efficient (p < 0.05). Echinacea sp. helps to reduce the incidence of mastitis-metritis-agalactia syndrome in pig herds (BOTH, 1987). The beneficial effects of E. purpurea supplementation on performance, weight gain and quality of meat showed that it can improve performance as well as maintain health by reducing the incidences of diarrhoea and disease outbreak by stimulating the immune system.

Supplementation of E. purpurea water extract (500 mg/kg of feed) in pig diets improved body weight gain and feed conversion, reduced cholesterol contents of meat and improved its lightness (HANCZAKOWSKA, 2007). Intermittent application (2 or 5 days) of E. purpurea juice followed by 12 or 9 days of interval improved health of pigs by increasing number of lymphocytes and leucocytes and increased phagocytosis rate of granulocytes (BHMER et al., 2008). The higher dose of extract (1000 mg/kg feed) improved water holding capacity of meat and raised it pH at 45 min after slaughter (HANCZAKOWSKA, 2007). KUHN et al. (2005) studied the effect of supplementation of E. purpurea ethanolic extract (0.125 ml/kg body weight) in liquid feed during the whole pregnant and suckling period of sows in six intervals (5 days treatment, followed by 2 weeks of break, respectively). The E. purpurea treatment of sows resulted in immunostimulatory effects both in sows and piglets with highest change in peripartal period. In one day old piglets, the concentrations of IgG, IgA, and CRP were significantly increased in E. purpurea group (P = 0.004, P < 000.1, P = 0.05, respectively). Up to 70 days of age, the rate of therapeutical treatments of piglets from E. purpurea treated sows was decreased in tendency (P = 0.08). The growth performance and carcass quality of offspring was not affected by E. purpurea treatment of sows. MAASS et al. (2005) conducted a number of experiments using dried cobs of E. purpurea herb as feed additive in diets of sows, piglets, and grower/finisher pigs on growth performance, blood picture, plasma enzymes including proliferation of lymphocytes, antibody status, and protein and immune globulin content of colostrum. The control groups were supplemented with alfalfa meal. The E. purpurea cobs consisted of aerial parts of E. purpurea plant, which were carefully dried and pressed. The sows received 0, 1.2, or 3.6% E. purpurea cobs in the diets continuously from day 85 to day 110 of gestation and 0, 0.5, or 1.5% E. purpurea cobs up to day 28 of lactation. In this experiment, no significant differences were found for growth performance, weight loss, blood picture, plasma enzymes, and colostrum composition. In second experiment, 1.8% E. purpurea cobs, or 20 mg/kg feed Flavomycin supplementation did not show any significant effect on performance and immune status. In third experiment, 48 grower/finisher pigs were used during a 9-wk experimental period with two supplementation phases (wk 13 and wk 79). The heat sterilised E. purpurea pressed juice obtained from aerial parts of the plant was used. The experimental groups received 0, 0.15% cobs or 46 ml pressed juice per day respectively. The juice was given on top of feed at both feeding times. E. purpurea supplemented groups showed significantly (P < 0.03) better feed conversion ratio than animals of the unsupplemented groups. In all these trials, the haematological parameters were not affected, but the rate of phagocytosis was enhanced. The authors concluded that dietary administration of E. purpurea in form of cobs as feed additive might have a stimulating effect on the immune system, especially in the situations with the increased stress for the immune system. Moreover, feed conversion can be improved with addition of E. purpurea to diets. HERMANN et al. (2003) conducted an experiment to study the effect of dietary E. purpurea on viremia and performance in porcine reproductive and respiratory syndrome virus (PRRSV)-infected nursery pigs. The certified organic E. purpurea ground root powder was mixed in basal diets at 2 and 4% and compared with a negative control and a diet containing carbadox (0.055 g/kg of diet; as fed basis) on PRRSV challenged pigs. Seven days after starting the diets all pigs were intranasally inoculated with PRRSV isolate (ATCC VR-2332) at a concentration of

Toxicity of Echinacea
The toxicity of Echinacea sp. appears to be very low. The results of experiments using different preparations and products of Echinacea sp. on various livestock species did not show any negative effects or signs of toxicity. There was no negative effect of E. purpurea juice supplementation on performance as well as liver functions in broilers (NASIR and GRASHORN, 2007a, 2007b). Amounts of lead and cadmium are very less in different preparation of E. purpurea (ZITKEVICIUS et al., 2003). MENGS et al. (1991, 2000) performed acute, sub-acute and genotoxicity studies on mice and rats and found E. purpurea to be virtually non-toxic to rats and mice. Single oral (15 g/kg body wt) or intravenous doses (5 g/kg body wt) of the expressed juice of E. purpurea proved virtually non-toxic to rats and mice. After four weeks of oral administration in doses amounting to many times the human therapeutic dose laboratory tests and necropsy findings gave no evidence of any toxic effects in rats. Tests for mutagenicity carried out in micro-organisms and mammalian cells in vitro and in mice all gave negative results. In an in vitro carcinogenicity study E. purpurea did not produce malignant transformation in hamster embryo cells. Studies looking for chromosome aberration and sister chromatid exchange in bacteria and cultured animal cells have also found no evidence of mutagenicity (MENGS et al., 1991). Maximum feasible oral and intravenous doses of ethanol-stabilized fresh pressed juice of E. purpurea similarly failed to cause measurable damage in mice or rats (MENGS et al., 2000). Application of E. purpurea root extract (2 mg/mouse/day) continuously from puberty (7 week) to 13 months did not show any signs of toxicity, rather it improved NK cells (BROUSSEAU and MILLER 2005). The squeezed sap of the E. purpurea plant is well tolerated in long term (up to 12 weeks) use, with no significant side effects when the sap was administered orally (PARNAHM, 1996). This conclusion was supported by HOBBS (1994) and KLIGLER (2003), who found no published reports indicating that Echinacea had toxic side
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Nasir and Grashorn: Echinacea as feed and water additive in livestock production
effects. E. purpurea contains saturated pyrolizidine alkaloids (isotussilagine and tussilage) at a level of 0.006%, which are also thought to be non toxic (PEARSON, 2000; NEWALL et al., 1996). CHOW et al. (2006) conducted an experiment in which pregnant mice were fed daily E. purpurea from pregnancy onset until gestational days 10, 11, 12, 13, and 14. A specific commercially prepared extract of E. purpurea in powdered form was homogenized into finally ground standard chow such that individual mice consumed 0.45 mg/day (dose per body weight). This chow was added to standard diet and offered at the beginning of the active period (6:00 pm). Controlled mice consumed identically prepared chow without E. purpurea. The results showed that the significant, pregnancy induced elevation in splenic lymphocytes and nucleated erythroid cells was all but eliminated in those females which consumed E. purpurea daily throughout their pregnancy. Moreover, consuming E. purpurea during pregnancy reduced the number of viable foetuses. High concentrations of E. purpurea have been shown to inhibit enzyme activity in human sperm, reduce oocyte penetration by sperm and can cause denaturation of sperm DNA (ONDRIZEK et al., 1999a; 1999b). The results of the experiments conducted using different preparations of E. purpurea suggest that there is no toxic or harmful effect on performance and health of laboratory animals as well as different livestock species. However, care should be taken, while feeding E. purpurea to animals kept for breeding purpose.

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ban imposed on the subtherapeutic use of antibiotics in animal feeds in European Union (EU) and the increasing demand of organic animal products, alternative substances are required, which can maintain health and improve performance of livestock without any residual effects. E. purpurea is one of the most promising phytogenic additives due to its immuno-stimulatory and potentiatory properties. Echinacea has been widely investigated in laboratory animals, as well as it is under continuous study in human trials for its potential clinical use. It contains a variety of active substances like alkamides, glycoproteins, polysaccharides, phenolic compounds, cinnamic acids, essential oils and flavonoids, which are effective in treatment of various ailments and improving immunity and health. The toxicity of Echinacea is reported to be very low. Present research on the use of Echinacea in livestock is limited, but it can be used as a guiding line and rationale for its use in livestock. In this review, the extent of present knowledge and need for future studies are discussed in the light of effectiveness of active substances of E. purpurea in improving health, performance and immunity of various livestock species.

Key words
Echinacea purpurea, swine, poultry, phytogenic
compounds, immunostimulator

Zusammenfassung Conclusions
The ban imposed on subtherapeutic use of antibiotics in animal feeds in EU, public pressure for banning antibiotics in other countries and increasing demand of organic animal products has created a whole new incentive and urgency to quantifying the usefulness of phytogenic products in animal nutrition. The outcome of experiments with phytogenic products may be used as guiding line and rationale for the application of these substances in livestock production. Echinacea is one of the few herbs that are extensively researched in laboratory animals as well as it is under continuous study in human trials for its potential clinical use. Present research on the use of E. purpurea in livestock is still limited but available results indicate positive effects in terms of weight gain, feed utilization and improved meat quality in poultry and swine. Stimulation of the immune system was also observed in cattle, horse, poultry and swine after oral (by feed or drinking water) administration of different E. purpurea products. But, as numerous Echinacea products are available in the market, differing from each other in their method of preparation and level of active substances, and as the mode of administration is not standardized a comparison of results in literature is difficult. Especially, the mode of administration (e.g. relation between application time and application free time; daily intake of active substances) is important for the probable effects on metabolism. Therefore, research is required to identify the most effective plant parts, level of inclusion, method and duration of application for poultry and swine in more detail.

Echinacea: Ein potentieller Futter- und Trinkwasserzusatz in der Geflgel- und Schweinerzeugung
Futterzusatzstoffe mit Leistung und Gesundheit steigernder Wirkung werden umfassend in der Tierftterung eingesetzt. ber einen langen Zeitraum waren Antibiotika die dominierenden Substanzen, da sie einen deutlichen Leistung steigernden Effekt aufwiesen. Inzwischen ist in der Europischen Union die Verwendung von Antibiotika in subtherapeutischen Dosen im Tierfutter zu Leistung steigernden Zwecken verboten. Das Verbot und auch die zunehmende Nachfrage nach kologisch erzeugten Tierprodukten erfordern in Zukunft alternative Substanzen, die die Aufrechterhaltung der Gesundheit und die Verbesserung der Leistung der Nutztiere ohne die Bildung von unerwnschten Rckstnden ermglichen. Echinacea purpurea ist auf Grund seiner das Immunsystem stimulierenden und steigernden Wirkung einer der am ehesten Erfolg versprechenden pflanzlichen Zustze. Echinacea wurde bisher am intensivsten bei Labortieren untersucht und wird auch in vielen Studien beim Menschen auf eine mgliche klinische Anwendung geprft. Echinacea enthlt viele verschiedene aktive Substanzen wie Alkamide, Glykoproteine, Polysaccharide, phenolische Komponenten, Zimtsure, essentielle le und Flavonoide. Diese gelten als sehr wirksam in der Behandlung von verschiedenen Leiden und verbessern sowohl die Immunitt als auch allgemein die Gesundheit. Es gibt bisher keine Erkenntnisse, die auf eine mgliche Toxizitt von Echinacea hindeuten. Die bisherigen Kenntnisse zum Einsatz von Echinacea bei landwirtschaftlichen Nutztieren sind begrenzt. Allerdings knnen die vorhandenen Informationen als Richtlinie fr den Einsatz in der Nutztierproduktion fungieren. In der vorliegenden bersicht wird daher der gegenwrtige Kenntnisstand zum Einsatz von Echinacea bei landwirtschaftlichen Nutztieren zusammengefasst und die Notwenigkeit fr weitere Untersuchungen im Hinblick auf die Effektivitt der aktiven Inhaltsstoffe von Echinacea purpurea bei der

Summary
Feed additives with performance and health stimulating effects are widely used in animal production. For a long time antibiotics have been the dominating substances in animal nutrition with distinct growth promoting effects. After the
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titis-metritis-agalactia syndrome of sows with natural drugs. Tierrztlische Umschau 11, 907-912. BHMER, B.M., H. SALISCH, B.R. PAULICKS, F.X. ROTH, 2008: Echinacea purpurea as a potential immunostimulatory feed additive in laying hens and fattening pigs by intermittent application. Livestock Science. In press: doi: 10.1016/j.livsci.2008.07.013. BROSSEAU, M., S.C. MILLER, 2005: Enhancement of natural killer cells and increased aging mice fed daily Echinacea root extract from youth. Biogeronthology. 6(3), 157-163. BURGER, R.A., A.R. TORRES, R.P. WARREN, V.D. CALDWELL, B.G. HUGHES, 1997: Echinacea-induced cytokine production by human macrophages. Intl. J. Immunopharm. 19, 371-379. CHOW, G., T. JOHNS, S.C. MILLER, 2006: Dietary Echinacea purpurea during Murine pregnancy: effect on maternal hemopoiesis and fetal growth. Biol. Neonate. 89, 133-138. CURRIER, N.L., S.C. MILLER, 2001: Echinacea purpurea and melatonin augment natural-killer cells in Leukemic mice and prolong life span. J. Alter. Compl. Med. 7(3), 241-251. CURRIER, N.L., S.C. MILLER, 2002: The effect of immunization with killed tumor cells, with/without feeding of Echinacea purpurea in an erythroleukemic mouse model. J. Alter. Compl. Med. 8(1), 49-58. DIETZ, B., J. HEILMANN, R. BAUER, 2001: Absorption of dodeca-2E, 4E, 8Z, 10E/Z tetraenoic acid isobutylamide after oral application of Echinacea purpurea tincture. Planta Med. 67, 863-864. FACINO, R.M., M. CARINI, G. ALDINI, L. SAIBENE, P. PIETTA, P. MAURI, 1995: Echinacea and Caffeoyl conjugates protect collagen from the free radical-induced degradation: a potential use of Echinacea extracts in the prevention of skin photo damage. Planta Med. 61, 510-514. FEDERICA, P., B. STEFANIA, M. Lara, M. MICHELE, S. FABRIZIA, 2003: Analysis of phenolic compounds and radical scavenging activity of Echinacea sp. Proc. 10th Intl meeting on recent developments in pharmaceutical analysis, Cogne, Italy, 289-301. GARDZIELEWSKA, J., K. PUDYSZAK, T. MAJEWSKA, M. JAKUBOWSKA, J. POMIANOWSKI, 2003: Effect of plant-supplemented feeding on fresh and frozen storage quality of broiler chicken meat. E. J. Pol. Agri. Uni. Anim. Husb. 6(2), http://www.ejpau.media.pl/volume6/issue2/animal/ art-12.html (accessed 03.05.2006). GILL, N.R., W. POWEL, S.T. FRANKLIN, K.I. MEEK, 2002: The effects of Echinacea on immune function of transitional calves. J. Anim. Sci. 80(Suppl. 1), 407. GOEL, V., C. CHANG, J.V. SLAMA, R. BARTON, R. BAUER, R. GAHLER, T.K. BASU, 2002: Echinacea stimulates macrophage function in the lung and spleen of normal rats. J. Nutr. Biochem. 13, 487-492. GROOM, S.N., T. JOHNS, P.R. OLDFIELD, 2007: The potency of immunomodulatory herbs may be primarily dependent upon macrophage activation. J. Med. Food. 10(1), 73-79. HANCZAKOWSKA, E., M. SWIQTKIEWICZ, 2007: Effect of supplementation with the purple coneflower (Echinacea purpurea) extract on fatty acid profile and quality of pig meat. Pol. J. Food Nutr. Sci. 57(4B), 229-233. HARBORNE, J.B., C.A. WILLIAMS, 2004: Phytochemistry of genus Echinacea. In: Echinacea: The genus Echinacea. S.C. Miller, Yu HE-CI (eds), CRC Press Washington, DC. HEINZER, F., M. CHAVANNE, J.P. MEUSY, H.P. MAITRE, E. GIGER, T.W. BAUMANN, 1988: Ein Beitrag zur Klassifizierung der therapeutisch verwendeten Arten der Gattung Echinacea. Pharm. Acta. Helv. 63, 132-136. HERMANN, J.R., M.S. HONEYMAN, J.J. ZIMMERMANN, B.J. THACKER, P.J. HOLDER, C.C. CHANG, 2003: The effect of
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Verbesserung der Gesundheit, der Leistung und der Immunitt bei verschiedenen Nutztierarten diskutiert.

Stichworte
Echinacea purpurea, Schwein, Geflgel, phytogene Inhaltsstoffe, Immunstimulation

Acknowledgements
Author Zahid Nasir was granted scholarship from Higher Education commission (HEC) of Pakistan and German Academic Exchange Service (Deutscher Akademischer Austausch Dienst DAAD) for his Ph D and research work at Hohenheim University, Stuttgart, Germany. We appreciate their financial support. Thanks goes to Berghof-Kruter GmbH, Heilsbronn, Germany for providing Echinacea purpurea juices.

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Correspondence: Dr. Zahid Nasir, Dept. of Poultry Science (470c), University of Hohenheim, Stuttgart 70599, Germany; E mail: Zahid_ch75@yahoo.com

Arch.Geflgelk. 4/2009