INFLUENCE OF SELENIUM COMPOUNDS ON GLUTATHIONE PEROXIDASE (GSH-PX) LEVEL IN LAMB BLOOD

Monika Hadryś¹, Stefania Kinal², Jolanta Antonowicz-Juchniewicz³, Iwona Jędrychowska^3
1 Department of Animal Nutrition and Feed Quality, Wrocław University of Environmental and Life Sciences, Poland
² Department of Animal Nutrition and Feed Science, Wrocław University of Environmental and Life Sciences, Poland
³ Department of Internal Medicine, Occupational Diseases and Hipertension, Wroclaw Medical University , Poland

ABSTRACT

The aim of the research was to measure the activity of GSH-Px in the blood of lamb being fed selenium in sodium selenite and selenium yeast to concentrate mixtures. The value of GSH-Px identified in the final stage of the experiment in all feeding groups was clearly higher that at the beginning (P < 0.01). Application of all selenium compounds used in the experiment to lambs in feed clearly influenced the increase of glutathione peroxidase (GSH-Px) activity in lamb blood (P < 0.01). The lambs receiving 0.3 mg Se/kg DM in feed in the form of sodium selenite and selenium yeast (groups I and III) showed higher (by 269.15 and 254.12 U/gHb respectively) glutathione peroxidase activity. The animals that were given selenium yeast in the amount of 0.2 mg Se/kg DM in feed, the activity of this enzyme was higher by 203.12 U/gHb.

Key words: glutathione peroxidase (GSH-Px),selenium, inorganic and organic forms, lambs.

INTRODUCTION

The physiological role of selenium that it plays in living organisms is mainly related to its strong impact on the growth and reproduction processes as well as developing specific and non – specific immunity level Undrewood and Suttle (1999), Grela (2001), Arthur et al.(2003) [2, 6, 18].

Present in selenocysteine, this microelement is an integral part of the active center of glutathione peroxidase (GSH-Px) – a system protecting cells from the impact of free radicals by becoming one of the most influential antioxidants. Glutathione peroxidase (GSH-Px) provides protection by reducing developed hydrogen and organic peroxides, as well as transforming them into water and adequate alcohols considerably less damaging to peroxides Friedrich (2002), Halliwell (1994), Undrewood and Suttle (1999) [5, 8, 18]. An adequate measure of involvement of selenium in metabolism is the activity of glutathione peroxidase (GSH-Px) in blood which reacts to the quantity and chemical form of selenium Harapin et al. (2000) [9].

Available literature contains no clear cut research data on the effect of application of various chemical forms of selenium on GSH-Px activity level in blood in animal feeding, particularly lambs. Therefore it appears important to carry out investigations in this field.

The aim of the research was to measure the activity of glutathione peroxidase (GSH-Px) in the blood of sheep being fed selenium in sodium selenite and selenium yeast to concentrate mixtures.

MATERIALS AND METHOD

The research material were 36 growing Polish Merino Sheep x Romanov Sheep crossbreed lambs with initial body weight of about 20 kg. They were kept in deep litter pens with unlimited access to water. The lambs were fed concentrate mixture and meadow hay in the amount of 1.0-1.2 and 0.2-0.5 kg/day/per head. The composition of concentrate mixture and feeding value of feeds given to the animals are presented in tables 1 and 2.

Table 1. Composition of concentrate mixture

Components	(%)
ground barley	37.0 %
ground wheat	30.0 %
soybean oil meal	10.0 %
rapeseed oil meal	10.0 %
wheat bran	10.0 %
fodder salt	0.3 %
limestone	0.7 %
premix	2.0 %

Table 2. Feeding value of concentrate mixture and meadow hay

Components (g)	Concentrate mixture	Meadow hay
– dry matter – crude protein – PDIN – PDIE – crude fat – crude fibre – crude ash – BAW – UFV – Calcium (Ca) – Phosphorus (P)	887.8 155 105 99 51.7 61.7 47.1 572.2 0.88 5.2 4.6	868.3 121.4 75.58 71.60 20.1 289.3 70.9 366.6 0.52 5.38 2.40

The lambs were assigned at random to 3 equal in number experimental groups (6 rams and 6 ewes in each group). The amount of selenium extracted from components of concentrate mixture equaled 0.13 mg Se/kg DM of feed The differentiating factor between the groups were various levels and forms of chemical bonds of selenium in mineral-vitamin mixtures being ingredients of concentrate mixtures for lambs.

Table 3. Scheme of experiment

Item	Selenium content (mg/kgDM)
	Group I	Group II	Group III
Feeds	0.13	0.13	0.13
Supplements:
– sodium selenite	0.3	-	-
– selenium yeast	-	0.2	0.3
Total	0.43	0.33	0.43

In control group I the lambs received selenium in the from of sodium selenite (0.3 mg Se/kg DM of feed), whereas in the experimental groups II and III, they were given selenium enriched yeast in the amount of 0.2 and 0.3 mg Se/kg DM of feed respectively. Selenium content in concentrate mixture for lambs ranged from 0.33 (group II) to 0.43 (group I i III) mg Se/kg DM of feed. Fattening was carried out until the animals’ body weight reached 35kg. Blood samples were taken from all lambs at the beginning and at the end of experiment from external carnival vein, the samples were collected before the morning meal. The activity of glutathione peroxidase (GSH-Px) was measured using Randox AOAC (2005) [1] tests in whole heparine-treated blood. The obtained values were statistically analysed using one-way variance analysis and the relevance of differences was estimated using the multiple range Duncan test Statistica (1998-2005) [17].

RESULTS

Figure 1 shows glutathione peroxidase (GSH-Px) activity in lamb blood. The level of glutathione peroxidase (GSH-Px) activity in lamb blood at the start of the experiment in respective groups (I, II, III) was diverse and amounted to 548.85 (± 78), 528.88 (± 27) and 532.17 (± 71) U/gHb respectively (Figure 1).

Fig. 1. Glutathione peroxidase (GSH-Px) activity in lamb blood

A,B – (P < 0.01)

The differences might be attributed to the individual variability of the animals. The level of glutathione peroxidase (GSH-Px) identified in the final stage of the experiment in all feeding groups was higher that at the beginning (Figure 1). Application of all selenium compounds used in the experiment to lambs in feed clearly influenced the increase of glutathione peroxidase (GSH-Px) activity in lamb blood (P < 0.01). The lambs receiving 0.3 mg Se/kg DM in feed in the form of sodium selenite and selenium yeast (groups I and III) showed higher (by 269.15 and 254.12 U/gHb respectively) glutathione peroxidase activity. The animals that were given selenium yeast in the amount of 0.2 mg Se/kg DM in feed, the activity of this enzyme was higher by 203.12 U/gHb.

DISCUSSION

Supplementation of selenium to lambs in feed in the organic (selenium yeast – 0.2 and 0.3 mg Se/kg DM) and inorganic forms (sodium selenite – 0.3 mg Se/kg DM) had a significant influence (P < 0.01) upon the increase in glutathione peroxidase activity in lamb blood. However, Podoll et al. (1992) in their study on lambs involving supplementation of inorganic selenium compounds in feed rations in the form of w sodium selenite and sodium selenate (0.3 mgSe/kg of feed mixture) found no influence of these forms of selenium upon GSH-Px activity in lamb blood [16]. Boldizarowa et al. (2005) found an increase in the GSH-Px activity in their study on evaluation of the influence of application of selenium enriched yeast (Selplex) for lambs in the amount of 0.3 mg Se/ kg DM. In contrast, GSH-Px activity did not increase in the animals receiving sodium selenite in feed mixture as well as in the blood of lambs from the control group that were not supplemented with selenium compounds [3]. Johansson et al. (1990) supplemented selenium yeast and sodium selenite in feed to lambs in the amount of 0.1 mg Se per day per head and found no increase in GSH-Px activity in the blood taken from these animals [10]. Person et al. (1989) in their research on heifers that were given selenium yeast, selenomethionine and sodium selenite in feed observed a twice as high increase in glutathione peroxidase activity in comparison with its level in the blood of heifers that received organic forms of selenium [15]. In the research on dairy cows that were given selenium yeast and sodium selenite in feed, Gunter et al. (2003) found an increase in GSH-Px activity in the blood of animals receiving selenium yeast in the perinatal and in calves delivered by such cows. The results related to GSH-Px activity in the blood of ruminants receiving both organic and inorganic forms of selenium are not in fact unambiguous in terms of the quantity as well as the chemical form of the selenium compounds supplemented [7]. However, GSH-Px activity in the blood of animals receiving organic forms of selenium shows a tendency to increase.

In the research on monogastric animals, a considerable controversy can be observed with respect to GSH-Px activity in the blood of these animals. Mahan et al. (1999) in their experiment on pigs consisting in supplementation of selenium yeast and sodium selenite in the amount ranging from 0.05 to 0.3 mg Se/kg DM observed a tendency to a lower GSH-Px activity in the blood of animals receiving the organic forms of selenium [13]. Mahan et al. (2004), in their research on sows receiving both selenium yeast (Selplex) and sodium in feed mixture in the amount of 0.15 and 0.3 mg Se/kg DM, found a similar increase in GSH-Px activity in blood [14]. Kim et al. (2001) in their research w on pigs consisting in supplementation in concentrate mixture various amounts of selenium (5, 10, 15 and 20 ppm) in the form of selenium enriched yeast and sodium selenite found no influence of these selenium compounds upon the level of GSH-Px activity in animal blood [11]. Leng et al. (2002), in their study on broiler chickens, applied feed mixtures with the content of 0.12 mg Se/kg selenium from natural components, which were supplemented with sodium selenite and selenium yeast (Selplex) to attain the level of 0.32 mgSe/kg and selenium yeast to attain the level of 0.82 mgSe/kg of the mixture. Their research showed that GSH-Px activity in the blood of chickens depended on the level of this micro-element in the feed ratio and not on the chemical form in which this element was present [12]. A similar dependency was also observed by Edens et al. (2004) in their research on broiler chickens [4].

CONCLUSIONS

The research data obtained indicate that the selenium compounds applied in the concentrate mixtures for growing lambs had a visible influence upon the increase of glutathione peroxidase in the blood of these animals. A higher increase in the level of GSH-Px activity was found in the blood of lambs that were given concentrate mixtures supplemented with sodium selenite and selenium yeast to attain the amount of 0.43 mg Se/kg DM. Such significant increase in the activity of this enzyme was not found in the lambs that were given selenium yeast to attain the level of 0.33 mg Se/kg DM. This may stand to indicate that GSH-Px activity in the blood of lambs depended on the quantity of the selenium yeast supplemented in the concentrate mixture containing selenium yeast.

REFERENCES

1. AOAC, 2005. Official Methods of Analysis of the Association on Official Analytical Chemists. 17^th Ed Kenneth, Helrich, Arlington, USA.

2. Arthur J.R., McKenzie R.C., Beckett G.J., 2003. Selenium in the immune system. J.Nutr.,133, pp. 1457-1459.

3. Boldizarowa K., Gresàkowà L.’, Faix S., Mellen M., Leng L’., 2005. Antioxidant status of lambs fed on diets supplemnted with selenite or Se-yeast. J.Anim.Feed Sci., 14, 245-253.

4. Edens F.W., Gowdy K.M., 2004. Selenium sources and selenoproteins in practical poultry production. Science and Technology in the Feed Industry. Proceedings of Alltech’s 20^th Annual Symposium. Edited by T.P. Lyons and K.A. Jacques. pp. 41-55.

5. Friedrich M., 2002. Składniki mineralne w żywieniu ludzi i zwierzat [Mineral Components in Human and Animal Nutrition]. Wydawnictwo Akademii Rolniczej w Szczecinie, pp. 69-71 [in Polish].

6. Grela E.R., 2001. Dodatki w żywieniu bydła [Supplements in Cattle Nutrition]. Praca zbiorowa, 30 [in Polish].

7. Gunter A., Beck P., Wistuba T., Davis M., Phillips J., 2003. effects of supplementary selenium source on performance, blood measurements, and immune function in beef cows and calves. Nutritional Biotechnology in the Feed and Food Industries. Proceedings of Alltech’s 19^th Annual Symposium, Edited by T.P. Lyons and K.A. Jacques, 295-308.

8. Halliwell B. 1994. Free radicals and antioxidants: A personal view. Nutr. Rev., 52, pp. 253-265.

9. Harapin I., Bauer M., Bedrica L., Potočnijak D., 2000. Correlation between gluthatione peroxidase activity and the quantity of selenium in the whole blood of beef calves. Acta Vet.,Brno, 69, pp. 87-92.

10. Johansson E., Jacobsson S.O., Luthman J., Lindh U., 1990. The biological response of selenium in individual erythrocytes and GSH-px in lambs fed sodium selenite or selenium yeast. J.Vet.Med. Assoc. 37, pp. 463-470.

11. Kim Y., Mahan D., 2001. Comparative effects of high dietary levels of organic and inorganic selenium on selenium toxicity of growing-finishing pigs. J. Anim. Sci. 79: 942-948.

12. Leng L., Bobcek R., Kuricovà S., Boldizarowa K., Gresàkowà L., Sevcíkovà Z., Rĕvajovà V., Levkutovà M., Levkut M., 2002. Comparative metabolic and immune responses of chickens fed diets containing inorganic selenium and Sel-Plex organic selenium. Nutritional Biotechnology in the Feed and Food Industries. Proceedings of Alltech’s 18^th Annual Symposium, Edited by T.P. Lyons and K.A. Jacques, 131-145.

13. Mahan D. C., Cline T. R., Richert B., 1999. Effect of dietary levels of selenium enriched yeast and sodium selenite as selenium sources fed to growing-finishing pigs on performance, tisssue selenium, serum glutathione peroxidase activity, carcass characteristics, and loin quality. J. Anim. Sci., vol. 77, Issue 8, pp. 2172-2179.

14. Mahan D., 2004. The role of selenium and Sel-Plex in sow reproduction.Science and Technology in the Feed Industry. Proceedings of Alltech’s 20^th Annual Symposium. Edited by T.P. Lyons and K.A. Jacques, pp.131-139.

15. Person B., Knutsson M., Gyllensward M., 1989. Gluthatione peroxidase activity in heifers fed diets supplemented with organic and inorganic selenium compounds. Swed J.Agr.Res. 19, pp. 53-56.

16. Podoll K., Bernard J., Ullrey D., DeBar S., Ku P., Magce W., 1992. Dietary selenate versus selenite for cattle, sheep and horses. J.Anim. Sci., vol. 70, Issue 6,1965-1970.

17. Statistica, 1998-2005. wersja 7.1, copyright StatSoft, Ins.

18. Underwood E.J., Suttle N. F., 1999. Mineral Nutrition of Livestock. Selenium. 3^rd editio. CAB International Publishing, New York, pp. 421-475.

 

Accepted for print: 6.03.2007

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Monika Hadryś
Department of Animal Nutrition and Feed Quality,
Wrocław University of Environmental and Life Sciences, Poland
J. Chełmonskiego 38d, 51-630 Wrocław, Poland
email: moniahad@interia.pl

Stefania Kinal
Department of Animal Nutrition and Feed Science,
Wrocław University of Environmental and Life Sciences, Poland
J. Chełmonskiego 38C, 51-630 Wrocław, Poland
phone: 71 320 58 29
email: stefania.kinal@up.wroc.pl

Jolanta Antonowicz-Juchniewicz
Department of Internal Medicine,
Occupational Diseases and Hipertension,
Wroclaw Medical University , Poland
Pasteur 4 Str, 50-367 Wrocław, Poland

Iwona Jędrychowska
Department of Internal Medicine,
Occupational Diseases and Hipertension,
Wroclaw Medical University , Poland
Pasteur 4 Str, 50-367 Wrocław, Poland

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