Daniel KORNIEWICZ, Henryk RÓŻAŃSKI, Zbigniew DOBRZAŃSKI 1, Piotr KACZMAREK, Adolf KORNIEWICZ 2

FOLIA UNIVERSITATIS AGRICULTURAE STETINENSIS Folia Univ. Agric. Stetin. 2007, Agric., Aliment., Pisc. Zootech. 257 (3), 81 90 Daniel KORNIEWICZ, Henryk RÓŻAŃSKI, Zbigniew DOBRZAŃSKI 1, Piotr KACZMAREK, Adolf KORNIEWICZ 2 THE EFFECT OF HERBIPLANT CS PHYTOPREPARATION ON HEMATOLOGICAL AND BIOCHEMICAL PARAMETERS IN YOUNG FATTENERS BLOOD WPŁYW FITOPREPARATU HERBIPLANT CS NA KSZTAŁTOWANIE SIĘ WSKAŹNIKÓW HEMATOLOGICZNYCH I BIOCHEMICZNYCH KRWI U MŁODYCH TUCZNIKÓW LNB Poland Ltd. Kiszkowo, 2/4 Rolna St., 62-280 Kiszkowo, Poland 1 Department of Animal Hygiene and Environment, Wrocław University of Environmental and Life Sciences, 38C Chełmońskiego St., 51-630 Wrocław, Poland 2 Department of Animal Nutrition and Feed Science, Wrocław University of Environmental and Life Sciences, 38D Chełmońskiego St., 51-630 Wrocław, Poland, e-mail: khz@ozi.ar.wroc.pl Abstract. Investigation involved the effect of phytopreparation administered in feed mixtures (125 kg 1 ) on hematological and biochemical blood parameters in young fatteners. The results proved that assessed preparation advantageously affected energy exchange (glucose concentration significantly increased) and lipid metabolism (significantly decreased concentration of total cholesterol, its LDL fractions and triglycerides).the preparation applied did not influence hemoglobin, erythrocyte content and hematocrit value, or the remaining indices of iron metabolism, yet it did significantly increase copper concentration. Perhaps longer application of phytopreparation and its higher doses in fattener s diet would provide their more diversified blood parameters. Key words: fatteners, hematological and biochemical parameters, phytopreparation. Słowa kluczowe: tuczniki, surowica krwi, wskaźniki hematologiczne i biochemiczne, fitopreparat. INTRODUCTION Withdrawing fodder antibiotics from animal feeding and increased consumer interest in so-called natural foods resulted in undertaking investigation on herb and medicinal plants in farm animals feeds (Grela et al. 1998; Stenzel et al. 2000; Paschma and Wawrzyński 2003; Korniewicz 2004; Kraszewski et al. 2004). Biologically active substances contained in herbs are of multidirectional effect: antibacterial, antifungal, immunomodifying (Grela et al. 1998; Ozcan and Boyraz 2000; Hart 2005; Korniewicz and Różański 2006), antioxidative and oxidative stress reducing in humans and animals (Yingming et al. 2004; Pajk et al. 2006). They influence numerous metabolic processes (Bhattaram et al. 2002), feature prophylactic and therapeutic effect, as well as can be treated as potential substitute for antibiotics (Hanczakowska and Urbańczyk 2002; Lien et al. 2007). According to Grela et al. (1998) herbs contain alcaloids, flavonoides polyphenoles, glicoproteides, polysaccharides, furanocumarines and lactones. According to Kamel (2001) and Korniewicz (2004) main active substances obtained from plants are cinnamon aldehyde, cineol, carvacrol, thymol, capsaicine, eugenol, anethol, allicine and menthol. These substances are of special considerable effects on increased activity of enzymes like amy-

82 D. Korniewicz et. al. lase, lipase, trypsin and enzymes from oxidcoreductase group like superoxide dismutase, glutathione peroxidase and catalase (Platel and Srinivasan 1996; 2000). These enzymes, removing free radicals, have an intestine villi protection, which in turn, increases their surface and possibilities of nutrients absorption. As a result, improvement of state of health and production effects become a fact, which was proved, among the others, by home investigation conducted by Kołacz et al. (1997), Grela et al. (1998), Grela (2000) and Korniewicz (2004). Literature concerning data herbs effect on the blood parameters: immunological, hematological and biochemical are very scarce. Some information can be found in the works of Kołacz et al. (1997) and Lien et al. (2007). The aim of investigation was to determine the effect of phytopreparation administered in feed mixtures on hematological and biochemical blood parameters in young fatteners. MATERIAL AND METHODS Phytopreparation was worked out in research-promoting centre LNB Poland Ltd. It is a mixture of micronized plant parts, volatile oils and plant extracts deposited on silicate and stabilized with palm oil. preparation is standarised regarding the content of the following active plant-originating substances: thymol 38 500, 1,8-cineol 35 000, carvacrol 10 000, pinene 4000, capsaicine 1700, cinnamon aldehyde 975, eugenol 450, flavonoids 6000, volatile oils 97 000 kg 1. in the form of powder was applied as a component of fodder mixtures supplementing Global Max (vitamin-mineral-amino acids) produced by LNB Poland Ltd. in Kiszkowo. Supplementary (vitamin-mineral-amino acids) mixtures, prepared according to the mentioned recipe, in the amount of 3 were introduced into feed mixtures. The same fodder materials were used in order to produce two kinds of mixtures differing only in the addition of preparation. Fatteners were classified into two nutrition groups: group I control, group II experimental with in the amount of 125 kg 1 feed mixture applied. Fodder materials used for feed mixtures production were subjected to chemical analysis in The Laboratory of The Department of Animal Nutrition and Feed Science at Wrocław University of Environmental and Life Sciences, using analytical methods in operation (AOAC, 1990). Energy value was calculated on the basis of component analysis and digestibility coefficient, as well as formulas inserted in Swine Feeding Standard (1993) and CVB (2004) Table 1. Ten hogs [(sow wbp x pbz) x boar (Hampshire x Pietrain)] of initial body weight of about 30 kg, classified into two nutrition groups were considered in the experiment. All animals were kept individually in pens equipped with pig feeders and nipple drinkers. The animals underwent digestion-balance examination, while blood examination were constituted additional on the experiment conducted in Experimental Animal Nutrition Plant in Gorzyń (Agricultural University in Poznań) and lasting out four weeks (Korniewicz et al. 2007). On the last day of investigation (28 th day in the morning before feeding) blood samples from jugular vein were collected from each fattener and the hematological indices (HGB, HCT RBC, MCV, MCH, MCHC, WBC, PLT), biochemical parameters (protein, albumins, glucose,

The effect of phytopreparation on hematological... 83 urea, liver enzymes, triglycerides, cholesterol and its fraction), as well as concentration of the following chemical elements: Ca, P, Mg, Na, K, Cl, Fe, Cu, Mn, Zn were evaluated. Table 1. Percentage composition and nutritive value of feed mixtures for fatteners Tabela 1. Skład procentowy i wartość pokarmowa mieszanek pełnoporcjowych dla tuczników Fodder ingredients Materiały paszowe Ground wheat Śruta pszenna Ground barley Śruta jęczmienna Ground triticale Śruta pszenżytnia Soya bean oilmeal Śruta poekstrakcyjna sojowa *MPU Farmer premix Global Max 1 Global Max 2 Units Jednostki miary I control kontrolna 20.00 42.00 20.00 15.00 Group Grupa II 20.00 42.00 20.00 15.00 3.00 3.00 Total Razem 100.00 100.00 In 1 kg of mixture W 1 kg mieszanki Metabolic energy Energia metaboliczna Nett energy Energia netto Dry matter Sucha masa Crude protein Białko ogólne Crude fiber Włókno surowe Crude fat Tłuszcz surowy Crude ash Popiół surowy Lysine Lizyna Methionine+Cystine Metionina + Cystyna Threonine Treonina Tryptophan Tryptofan Isoleucine Izoleucyna Ca total Ca razem P total P razem P digestible P przyswajalne Na Mg Zn Fe Cu Mn MJ kcal 12.90 2220 86.74 15.90 2.91 1.30 4.85 1.02 0.20 0.67 0.54 0.32 0.18 0.11 172 178 20 50 12.90 2220 86.74 15.90 2.91 1.30 4.85 1.02 0.20 0.67 0.54 0.32 0.18 0.11 172 178 20 50 125 *MPU also contain xynalase, gluconase and phytase MPU zawiera też ksynalazę i glukanazę oraz fitazę. Analysis in The Laboratory of Department of Animal Hygiene and Environment at Wrocław University of Environmental and Life Sciences was conducted, using biochemical analyser Pentra 400 produced by Horiba ABX. For Cu and Zn analysis RANDOX firm reagents were used, while Mn concentration in serum was evaluated according to AAS method, applying microwave sample mineralization. Hematological examinations of samples evaluated were done with the use of hematological analyser ABC-Vet. The results of analysis were statistically worked out using one-factor variance analysis and difference significance between the groups was estimated according to t -Student test (Statgraphics v. 5.0 software).

84 D. Korniewicz et. al. RESULTS AND DISCUSSION Mixtures assessment Feed mixtures for both groups (Table 1) contained 15.9 crude protein and 1.02 crude lysine. The remaining balanced aminoacids like methionine, cystine, threonine, tryptophan and isoleucine were in even amounts. Protein and amino acids levels, macroelements (Ca, P, Mg, Na) and microelements content (Fe, Cu, Mn and Zn) was even in both mixtures and followed with recommendations included in polish Swine Feeding Standard (1993) and with the pig feeding programme by LNB Poland Sp. Ltd. worked out by Korniewicz (2006). The results of fattening, digestibility of basic nutrients, protein balance, apparent absorption and macroelements (Ca, P i Mg) and microelements (Fe, Cu, Mn I Zn) balances are the subject of separate elaboration (Korniewicz et al. 2007). Hematological indicators The results of hematological examination (Table 2) reveal that applied preparation did not affect hemoglobin concentration (HGB), hematocrite (HCT) and erythrocyte concentration (RBC). Table 2. Hematological indicators of fatteners blood Tabela 2. Wskaźniki hematologiczne krwi tuczników Specification Wyszczególnienie Hemoglobin (HGB) Hemoglobina [mmol l 1 ] Hematocrit Hematokryt (HCT) [] Red blood cells Erytrocyty (RBC) [10 9 l 1 ] Mean corpuscular volume Średnia objętość erytrocytów (MCV) [10 9 l 1 ] Mean corpuscular hemoglobin Średnia zawartość hemoglobiny w erytrocytach (MCH) [pg] Mean corpuscular hemoglobin concentration Średnie stężenie hemoglobiny w erytrocytach (MCHC) [mmol l 1 ] White blood cells Leukocyty (WBC) [10 9 l 1 ] Platelets Płytki krwi (PLT) [10 9 l 1 ] I control kontrolna Group Grupa II 11.04±0.99 11.35±0.57 38.30±3.95 39.05±1.52 6.56±0.57 6.72±0.08 58.0±1.67 58.0±2.86 16.68±0.61 16.92±0.91 28.70±0.51 28.97±0.38 23.16±4.76 23.85±3.75 322 ± 28.2 324 ± 24.4 Mean marked with the different superscript letters differ significantly: a,b p 0.05 and A,B p 0.01. Średnie wartości oznaczone różnymi literami różnią się istotnie: a,b p 0,05 oraz A,B p 0,01. Any differences between control and experimental groups regarding the remaining indices connected with iron metabolism, such as mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC)

The effect of phytopreparation on hematological... 85 were not recorded. White blood cells (WBC) and platelets (PLT) were equal in both nutrition groups. Controlled hematological indicators remained within the range of reference values by Winnicka (1997) and they were similar to those obtained by Korniewicz et al. (2000). Investigation by Lien et al. (2007) revealed that 8-week application of Chinese herbs Bazhen in weaned piglets significantly increased WBC (from 7.46 to 11.09 x 10 3 m 1, RBC (from 6.28 to 6.69 x 10 9 l 1 ) and some immunological indicators like gammaglobulins, including immunoglobulin G. Any alterations in concentrations of MCV, MCHC, albumins and globulins were recorded. Other authors (Yu et al. 2000) also observed the increase in some hematological indicators (HGB, PCV) in piglets receiving aminoacid-iron preparation ( Availa ). Hematological indicators alterations are especially characteristic in young pigs. Apart from technological-environmental factors (e.g. time of weaning, the way of rearing, temperature of the environment), the most important is the way of feeding and fodder quality, including the content of available iron (Link and Kovač 2000). Animal state of health, particularly parasitic diseases or diarrhea occurrence also affect on blood parameters in pigs (Wieczorek et al. 2006). Biochemical parameters Investigation results regarding biochemical parameters in weaned piglets blood serum (Table 3) point to the effect of phytopreparation administered in feed mixture. Table 3. Biochemical parameters of fatteners blood serum Tabela 3. Wskaźniki biochemiczne w surowicy krwi tuczników Specification Wyszczególnienie I control kontrolna Group Grupa II Total protein Białko całkowite [g l 1 ] 67.7±4.29 65.3±5.91 Albumins Albuminy [g l 1 ] 55.6±6.04 52.4±3.27 Urea Mocznik [mmol l 1 ] 4.34±0.70 4.18±0.68 Glucose Glukoza [mmol l 1 ] 2.26 A ±0.53 4.17 B ±0.70 AST [U l 1 ] 56.6±14.97 51.7±14.47 ALT [U l 1 ] 74.03±4.02 62.1±3.50 GGT [U l 1 ] 45.6±11.62 49.1±13.10 Cholesterol Cholesterol [mmol l 1 ] 2.63 a ±0.12 2.33 b ±0.26 HDL [mmol l 1 ] 0.81±0.08 0.80±0.06 Triglyceride Trójglicerydy [mmol l 1 ] 0.42 a ±0.03 0.36 b ±0.05 LDL [mmol l 1 ] 1.62 a ±0.10 1.37 b ±0.22 Mean marked with the different superscript letters differ significantly: a,b p 0.05 and A,B p 0.01. Średnie wartości oznaczone różnymi literami różnią się istotnie: a,b p 0,05 oraz A,B p 0,01. Yet did not effect on such indices of protein metabolism as: concentration of total protein, albumins or urea. There were not recorded any significant differences between the groups regarding enzyme activity or alanine and asparagine transaminase. Nev-

86 D. Korniewicz et. al. ertheless, significantly (p 0.01) increased glucose concentration from 2.26 to 4.17 mmol l 1 can prove advantageous effect of this phytopreparation on energy balance in fatteners. Similar influence of herbs on fat metabolism in fatteners was recorded by Grela (2000). There was also proved favourable effect on the assessed preparation on lipids metabolism, which can be confirmed by significant decrease in crude cholesterol level, its LDL fraction and triglycerides. This fact is difficult to be interpreted because of complicated mechanisms of lipids metabolism in pigs and other animals. Another investigation by Korniewicz et al. (2007) involving administration of yeast Saccharomyces cerevisiae enriched in bioelements (Fe, Cu, Mn) to weaned piglets also confirmed the effect of microelements in an organic form on decreased cholesterol concentration in blood serum. Biochemical parameters in pigs, especially piglets and weaned piglets can vary within certain limits, which was observed by Świtała et al. (1998) and Migdał et al. (2003). Mineral components in blood serum Obtained results of blood serum analysis in the view of mineral components are showed in Table 4. Table 4. Concentration of mineral components in pig blood serum Tabela 4. Stężenie składników mineralnych w surowicy krwi Specification Wyszczególnienie I control kontrolna Group Grupa II Ca [mmol l 1 ] 2.57±0.14 2.67±0.41 P [mmol l 1 ] 3.55±0.40 3.57±0.27 Mg [mmol l 1 ] 1.00±0.10 1.00±0.04 Na [mmol l 1 ] 136.4±1.91 135.7±1.25 K [mmol l 1 ] 5.78±0.97 5.07±0.29 Cl [mmol l 1 ] 105.4±2.03 105.2±0.96 Fe [μmol l 1 ] 20.02±3.50 20.34±1.87 Cu [μmol l 1 ] 35.50 a ±1.61 40.59 b ±4.62 Mn [μmol l 1 ] 0.48±0.06 0.52±0.05 Zn [μmol l 1 ] 34.82±4.16 37.92±10.26 Mean marked with the different superscript letters differ significantly: a,b p 0.05 and A,B p 0.01. Średnie wartości oznaczone różnymi literami różnią się istotnie: a,b p 0,05 oraz A,B p 0,01. They prove that preparation administered in fatteners fodder did not effect on the concentration of the examined macroelements, i.e. Ca, P, Mg, Na, K, Cl. Determined concentration of these mineral components is similar to reference values reported by Winnicka (1997) and the ones obtained in previous investigation by Korniewicz et al. (2003). No significant differences were found between the groups regarding iron or manganese concentration. Nevertheless, it was possible to prove that preparation assessed did significantly (p 0.05) effect on increased copper concentration, which suggests its improved availability.

The effect of phytopreparation on hematological... 87 The data involving the effect of herb preparations on concentration of chemical elements in pig blood serum have not been found. Concentration of mineral components in pig serum varies relatively and depends on elements content in animal diet, as well as their bioavailability. Revy et al. (2004) reported significant increase in Zn in blood serum without any effect on P and Cu concentration in weaned piglets receiving microbiological phytase with zinc. Nowotný et al. (2005) administered in weaned piglets fodder the following elements: Fe, Cu, Mn, Zn and Se in an organic or traditional form. There were controlled concentrations of those elements in blood serum collected from animals aging 65, 72, 80 and 93 days. In all periods significant effect on organic form on increased concentration of microelements content could be recorded: Fe from 14.66 to 18.29; Cu from 17.8 to 22.71; Zn from 8.50 to 9.82; Mn from 0.43 to 0.57μmol l 1. Czech and Grela (2006), administering chelates Biomax to fatteners, observed significant increase in Cu, Mn and Zn concentration in blood serum, while no effect was recorded in the case of Ca, P, Mg and Fe concentration or biochemical indices. CONCLUSIONS 1. phytopreparation administered in feed mixtures (125 kg 1 ) affected on energy exchange (glucose concentration significantly increased) and lipid metabolism (significantly decreased concentration of total cholesterol, its LDL fractions and triglycerides). 2. The preparation applied did not effect on content of hemoglobin, erythrocyte and hematocrite value, as well as the remaining indices of iron metabolism, yet it did significantly increase in blood serum of copper concentration. 3. Probably, long-term administration of phytopreparation and its doses in animal diet would provide more diversified picture of fatteners blood parameters. REFERENCES AOAC. 1990. Official methods of analysis of the Association of Official Analytical Chemists. Arlington, Virginia, USA. Bhattaram V.A., Graefe U., Kohlert C., Veit M., Derendorf H. 2002. Pharmacokinetics and Bioavailability of Herbal Medicinal Products. Phytomedicine 9, 1 33. CVB. 2004. Veevoedertabel. Centraal Veevoederbureau, Lelystad, The Netherlands. Czech A., Grela E.R. 2006. Wpływ dodatku chelatów mineralnych w mieszankach dla rosnących świń na wzrost i składniki krwi [The effect of suplement mineral chelates for growing pigs on growth and blood parameters]. Ann. UMCS, Sect. EE 16, 111 117 [in Polish]. Grela E.R. 2000. Influence of herbs mixture in pigs feeding on performance of some lipid parameters in blood and backfat. Ann. UMCS, Sect. EE 18, 245 250. Grela E.R., Sembratowicz L., Czech A. 1998. Immunostymulacyjne działanie ziół [Immunostimmulatory activity of herbs]. Med. Weter. 54, 152 158 [in Polish]. Hanczakowska E., Urbańczyk J. 2002. Efficiency of herb mixtures as antibiotic replacers for piglets according to their age. Ann. Anim. Sci. 2, 131 138.

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