Carbohydrate and protein metabolism predominate during the first two weeks of incubation, with the last seven days of incubation being noted as a period of intense lipid metabolism and rapid embryo growth in which some 80% of the entire lipid content is mobilized and absorbed into the embryonic tissue (Romanoff, 1960; Noble and Cocchi, 1990). Fatty acid oxidation is the primary source of energy and metabolic water for developing embryos (Rahn and Paganelli, 1991). Part of this lipid material is composed of unsaturated fatty acids, which are prone to peroxidation (Surai, 2002).
The beneficial effect of Se during embryo development was verified by Paton et al. (2002). First of all, total egg Se content is higher when Se is provide to the hen in organic form. Organic Se as Sel-Plex® also enhanced the antioxidant profile of the embryo (Surai and Sparks, 2000). Therefore, incorporation of Se (as a component of GSH-Px) by the body of the chick brings about protection against oxidation reactions.
In addition, there is an increase in conversion of linoleic acid to arachidonic acid, an important component of cell membrane phospholipids. That fatty acid is unsaturated and therefore needs protection from oxidizing reactions. Recently, Pappas et al. (2004) indicated that dietary fish oil (or PUFAs) may affect Se metabolism and deposition in the tissues rather than Se absorption from the diet. Fish oil may therefore influence uptake mechanisms for Se in the embryo.
Selenium status is also critical to hematopoiesis (Paton et al., 2002). On day 12 of incubation, the spleen begins to function as a hematopoietic organ and produces primitive lymphoid cells that differentiate into leucocytes and erythrocytes. The development of erythrocytes would require additional synthesis of GSH-Px. Finally, Paton and coworkers mentioned that the adenohypophyseal-thyroid axis (embryonic thyroid) of the chick becomes functional on day 11.5 of incubation and the thyroid of the chick begins to convert thyroxin to triiodothyronine, the active form of thyroid hormone. This conversion depends on deiodinase enzymes, which are Se-dependent.
The added Bioplex™ organic Zn and Mn may have helped in embryo development and survival during the late part of incubation. Vieira and Moran (1998) compared mineral content of 27 and 62-week old broiler breeder hens and indicated that there is a reduction in yolk Zn and Mn content in older hens. Feeding layer (Guo et al., 2002) and broiler breeder (Hudson et al., 2004a) hens diets containing organic Zn increased egg Zn content. In addition, Badawy et al. (1987) found a positive relationship between Zn concentration of eggs and hatchability. Therefore, Hudson et al. (2004a) hypothesized that inadequate transmission of Zn from the hen to the hatching egg is likely responsible for low hatchability and poor chick quality when Zn intake is insufficient.
According to Hudson et al. (2004a), organic and inorganic Zn in combination increase the number of settable eggs per hen housed, and thus the total number of chicks produced, but not fertility or hatchability. The authors proposed two mechanisms of action for better performance due to the combination of organic and inorganic Zn sources as opposed to either fed as the sole source of dietary Zn. First, absorption of Zn and Zn status of the bird may be improved.
Supplementing a diet with both organic and inorganic sources of Zn may involve more absorption sites or transporters in the intestine, increasing Zn retention and enhancing performance. In addition, chelation of Zn by phytic acid in the gastrointestinal tract may be reduced and absorption is likely increased when organic Zn sources are provided in the diet. A second possible mode of action may involve effects of dietary Zn source on Zn metabolism. Dietary Zn from organic sources may be absorbed intact and function differently than Zn from inorganic source after being absorbed. Suso and Edwards (1972) suggested that Zn may be transferred to enzymes as a metalloligand complex, and the ligand could influence enzyme activity.
Zinc is also involved in immune status (Leeson and Summers, 2001). Hudson et al. (2004a) have shown that organic Zn enhanced the immune status of hens, indicating that disease resistance or transmission of antibodies to progeny may be enhanced when broiler breeder hens consume organic Zn. However, in their study that effect was not translated into lower embryonic mortality. In addition, supplemental Zn from an organic or inorganic source in breeder hen diets did not influence chick physiology at hatching (Hudson et al., 2004b).
PERFORMANCE OF PROGENY
According to Lopez and Leeson (1994), the biological function of the broiler breeder is to produce a viable embryo with the features required to produce a broiler chicken. A fertile egg will provide a closed environment within which all nutritional needs of the embryo must be met, with the notable exception of gaseous change. Therefore, the physiological condition of the broiler breeder hen and the egg are related to normal embryo development.
A high positive relationship between egg weight and chick size has been reported by several workers (Shanawany, 1987; Pinchasov, 1991) and chick weight is usually 62-76% of egg weight (Lopez and Leeson, 1994). Chick weight at hatch affects subsequent broiler growth. It has been reported that a 1 g difference in egg weight results in 0.5 g in chick size, which subsequently results in an approximate 5 g difference in body weight at 42 days of age (Leeson and Summers, 2000). Higher broiler weight at slaughter is the main goal of the farmer. Egg size increases with breeder age, so it would be expected that bird age will also affect chick weight.
Dietary manipulation of certain nutrients during the latter parts of the growing period and the early stages of the egg production cycle has the potential to increase early egg size. Previous investigations have shown that early egg size can be increased by increasing dietary protein during the early stages of egg production (Keshavarz, 1995). Changes in protein and amino acid content of the diet primarily affect the deposition of albumen (Etches, 1996; Joseph et al., 2000). The benefit of higher dietary protein on early egg size can be further increased by supplemental fat in isocaloric diets (Keshavarz and Nakajima, 1995). It is important to point out that changes in the consumption of linoleic acid alter yolk formation (Etches, 1996).
Replacement of inorganic Se with Sel-Plex® has been shown to increase both the yolk and albumen weight (Pan et al., 2004; Xavier et al., 2004) as well as the dry matter content of the yolk (Lara, personal communication). Because chick size is positively correlated with egg size, it seems reasonable to assume that changes in the diet that influence egg weight will also influence the size of the chick at hatch and subsequent performance. Surai (2000) observed higher GSH-Px activity in chicks from breeders fed Sel-Plex® than in those fed sodium selenite. This fact certainly explains the lower culling and mortality of chicks from hens fed Sel-Plex® (Lanning et al., 2000). Furthermore, these data are corroborated by results of Rebel et al. (2004), who reported that additional vitamins and trace minerals (Se, copper and Zn) in breeder diets can stimulate the immune system of chicks.
ALBUMEN CONSISTENCY AND INCUBATION: POSSIBLE ROLE OF SEL-PLEX® FOR DELAYED INCUBATION?
After oviposition, there is a rise in albumen pH with storage time, and hen age is associated with a decrease in albumen height and viscosity. Albumen liquefaction probably facilitates the movement of nutrients from the albumen to the blastoderm (Brake et al., 1997) and may reduce resistance to gaseous diffusion (Meuer and Baumann, 1988). This explains why short-term storage may have beneficial effects on hatchability of eggs from young flocks (Brake, 1996), as these flocks generally lay eggs that have albumen of good quality (density) that is fairly resistant to degradation. However, extended periods of egg storage allow the albumen to degrade excessively. This degradation causes the blastoderm to move into close proximity to the egg shell, so that early embryonic mortality results from dehydration during the early stages of incubation (Brake et al., 1993). Lapão et al. (1999) have shown that in older breeder flocks, the decline in hatchability starts 1 day after lay, possibly due to deterioration in egg albumen quality.
The detrimental effect of long-term storage is more pronounced in eggs from older breeder flocks (Kirk et al., 1980), which is a result of lower albumen quality at oviposition and a consequent increased rate of decline during storage (Hurnik et al., 1978). Elibol and Brake (2004) observed that fertile egg hatchability was higher from a broiler breeder flock at 29 weeks of age than at 68 weeks of age because of decreased mortality at all stages of embryo development. Recently, Tona et al. (2004) confirmed those observations and indicated that fresh eggs from young breeders had better albumen quality, hatched better, and produced a higher percentage of high quality chicks, although with lower weights at hatch, compared with older breeders but showed greater post-hatch growth rate.
The low egg albumen quality effect has been shown to be alleviated by adding Sel-Plex® to the breeder diets in several studies. Examining the effect of Sel-Plex® on albumen consistency over one week (Wakebe, 1998), two weeks (Pan et al., 2004) or a combination of organic minerals containing Sel-Plex® over 12 days (Xavier et al., 2004), an improvement in the quality of albumen through dietary Sel-Plex®, as measured by Haugh units, has been shown.
The beneficial effect of Sel-Plex® on albumen height represents protection of cellular membranes of the magnum, which are basically formed by cholesterol, phospholipids and peripheral and integral proteins (Vander et al., 1990). One of the roles of integral proteins is to serve as nutrient carriers to and from extracellular to intracellular fluid. Therefore, one may hypothesize that by protecting the membrane of secretory cells and tubular glands of the magnum, Se as a component of GSH-Px allows the secretory granules to discharge their contents more efficiently. The more protein gets into the magnum lumen, the more viscous the egg white becomes (Butts and Cunningham, 1972; Joseph et al., 2000). It should be noted that histological characteristics of the cells in the tubular glands indicate that they produce egg white proteins continuously throughout the ovulatory cycle and store the proteins in secretory granules (Etches, 1996).
As mentioned above, a reduction in albumen is observed when birds receive low-protein diets, suggesting that these diets are lower in essential amino acids (Butts and Cunningham, 1972; Joseph et al., 2000). Those proteins are produced in the rough endoplasmic reticulum and packaged by the Golgi apparatus. Then, proteins are transferred into secretory granules until the moment they are secreted into the magnum (Sandoz et al., 1971). Since rough endoplasmic reticulum and Golgi apparatus are cell organelles surrounded by membranes, membranes must be intact for proper function. This again indicates an important role of organic Se in improving Se status and consequently antioxidant status.
Conclusions
The beneficial effects of organic mineral supplements on broiler breeder performance are very consistent. Favorable responses have been observed at various ages of supplementation, however, it is advised to begin supplementation during the pre-lay diet and continue throughout the reproductive cycle.
References
Arnold, R.L., O.E. Olson and C.W. Carlson. 1974. Tissue selenium content and serum tocopherols as influenced by dietary type, selenium and vitamin E. Poult. Sci. 53:2185-2192.
Badawy, E.M., B.M. Edrise and A.M. Al-Wakeel. 1987. The relationship between quality, egg constituents and hatchability of the eggs of Hubbard broiler breeders:
a field study. Vet. Med. J. 35:105-115.
Barbato, G.F. 1999. Genetic relationships between selection for growth and reproductive effectiveness. Poult. Sci. 78:444-452.
Bekhtina, V.G. 1968. Morphological features of polyspermy fecundation in hens. In: Summaries from Pushkin Research Laboratory of Livestock Breeding. Leningrad Region, USSR. Cited by World’s Poult. Sci. J. 24:148.
Brake, J.T. 1996. Optimization of egg handling and storage. World Poult. Misset 12(9):33-39.
Brake, J., J.D. Garlich and E.D. Peebles. 1985. Effect of protein and energy intake by broiler breeders during the prebreeder transition period on subsequent reproductive performance. Poult. Sci. 64:2335-2340.
Brake, J., T. Walsh and S.V. Vick. 1993. Hatchability of broiler eggs as influenced by storage and internal quality. Zootech. Int. 16(1):30-41.
Brake, J., T.J. Walsh, C.E. Benton, J.N. Petitte, R. Meiherhof and G. Penalva. 1997. Egg handling and storage. Poult. Sci. 76:144-151.
Bramwell, R.K. and B. Howarth, Jr. 1992. Preferential attachment of cock spermatozoa to the perivitelline layer directly over the germinal disc of the hen’s ovum. Biol. Reprod. 47:1113-1117.
Bramwell, R.K., H.L. Marks and B. Howarth, Jr. 1995. Quantitative determination of spermatozoa penetration of the perivitelline layer of the hen’s ovum as assessed on oviposited eggs. Poult. Sci. 74:1875-1883.
Brillard, J.P. 2004. Natural mating in broiler breeders: present and future concerns. World’s Poult. Sci. J. 60:439-445.
Butts, J.N. and F.E. Cunningham. 1972. Effect of dietary protein on selected properties of the egg. Poult. Sci. 51:1726-1734.
Cantor, A.H. 1997.The role of selenium in poultry nutrition. In: Biotechnology in the Feed Industry, Proceedings of Alltech’s 13th Annual Symposium (T.P. Lyons and K.A. Jacques, eds). Nottingham University Press, UK, pp. 155-164.
Cave, N.A.G. 1984. Effect of a high-protein diet fed prior to the onset of lay on performance of broiler breeder pullets. Poult. Sci. 63:1823-1827.
Edens, F.W. 2002. Practical applications for selenomethionine: broiler breeder reproduction. In: Nutritional Biotechnology in the Feed and Food Industries, Proceedings of Alltech’s 18th Annual Symposium (T.P. Lyons and K.A. Jacques, eds). Nottingham University Press, UK, pp. 29-42.
Elibol, O. and J. Brake. 2004. Identification of critical periods for turning broiler hatching eggs during incubation. Br. Poult. Sci. 45:631-637.
Eslick, M.L. and G.R. McDaniel. 1992. Interelationships between fertility and hatchability of eggs from broiler breeder hens. J. Appl. Poult. Res. 1:156-159.
Etches, R.J. 1996. Reproduction in Poultry. 1st ed., CAB International, Wallingford, UK.
Fontana, E.A., W.D. Weaver, Jr. and H.P. van Krey. 1992. Intermittent periods of infertility identified in naturally mated broiler breeder hens. J. Appl. Poult. Res. 1:190-193.
Guo, Y.M., R. Yang, J. Yuan, T.L. Ward and T.M. Fakler. 2002. Effect of AvailaZn and ZnSO4 on laying hen performance and egg quality. Poult. Sci. 81(Suppl. 1):40.
Gyles, N.R. 1989. Poultry, people and progress. Poult. Sci. 68:1-8.
Hammerstedt, R.H. 1999. Symposium summary and challenges for the future. Poult. Sci. 78:459-466.
Havenstein, G.B., P.R. Ferket and M.A. Qureshi. 2003a.
Growth, livability and feed conversion of 1957 versus 2001 broilers when fed representative 1957 and 2001 broiler diets. Poult. Sci. 82:1500-1508.
Havenstein, G.B., P.R. Ferket and M.A. Qureshi. 2003b. Carcass composition and yield of 1957 versus 2001 broilers when fed representative 1957 and 2001 broiler diets. Poult. Sci. 82:1509-1518.
Hazary, R.C., H.J. Staines and G.J. Wishart. 2000. Assessing the mating efficiency and predicting fertility in broiler breeder flocks by quantifying spermatozoa which penetrate the inner perivitelline layer over the germinal disc. Br. Poult. Sci. 41:395-400.
Hudson, B.P., W.A. Dozier, J.L. Wilson, J.E. Sander and T.L. Ward. 2004a. Reproductive performance and immune status of caged broiler breeder hens provided diets supplemented with either inorganic or organic sources of zinc from hatching to 65 wk of age. J. Appl. Poult. Res. 13:349-359.
Hudson, B.P., B.D. Fairchild, J.L. Wilson, W.A. Dozier and J. Buhr. 2004b. Breeder age and zinc source in broiler breeder hen diets on progeny characteristics at hatching. Poult. Sci. 13:55-64.
Hurnik, G.L., B.S. Reinhart and J.F. Hurnik. 1978. Relationship between albumen quality and hatchability in fresh and stored eggs. Poult. Sci. 57:854-857.
Jacques, K.A. 2001. Selenium metabolism in animals: the relationship between dietary selenium form and physiological response. In: Science and Technology in the Feed Industry, Proceedings of Alltech’s 17th Annual Symposium (T.P. Lyons and K.A. Jacques, eds), Nottingham University Press, UK, pp. 319-348.
Jensen, L.S. 1968. Selenium deficiency and impaired reproduction in Japanese quail. Proc. Soc. Exp. Biol. Med. 128:970-972.
Joseph, N.S., F.E. Robinson, D.R. Korver and R.A. Renema. 2000. Effect of dietary protein intake during the pullet-to-breeder transition period on early egg weight and production in broiler breeders. Poult. Sci. 79:1790-1796.
Kerr, C.L. 1998. Genetics of female-related fertility in chickens. PhD Dissertation. The Pennsylvania State University, University Park, PA.
Keshavarz, K. 1995. Further investigations on the effect of dietary manipulations of nutrients on early egg weight. Poult. Sci. 74:62-74.
Keshavarz, K. and S. Nakajima. 1995. The effect of dietary manipulations of energy, protein and fat during the growing and laying periods on early egg weight and egg components. Poult. Sci. 74:50-61.
Kirk, S.G., G.C. Emmans, R. McDonald and D. Arnot. 1980. Factors affecting the hatchability of eggs from broiler breeders. Br. Poult. Sci. 21:37-53.
Kuroki, M. and M. Mori. 1997. Binding of spermatozoa to the perivitelline layer in the presence of a protease inhibitor. Poult. Sci. 76:748-752.
Lanning, D., K. Ayres and S. Kenyon. 2000. Sel-Plex® in the breeder diet: reductions in chick mortality: summary of commercial studies in Britain 2000.
Alltech UK, Stampford, Lincs, UK. Lapão, C., T.L. Gama and M.C. Soares. 1999. Effects of broiler breeder age and length of egg storage on albumen characteristics and hatchability. Poult. Sci. 78:640-645.
Leeson, S. and J.D. Summers. 1997. Commercial Poultry Nutrition. 2nd ed., University Books, Guelph, Ontario, Canada.
Leeson, S. and J.D. Summers. 2000. Broiler Breeder Production. 1st ed., University Books, Guelph, Ontario, Canada.
Leeson, S. and J.D. Summers. 2001. Scott’s Nutrition of the Chicken. 4th ed., University Books, Guelph, Ontario, Canada.
Lopez, G. and S. Leeson. 1994. Nutrition and broiler breeder performance: a review with emphasis on response to diet protein. Appl. Poult. Res. 3:303-311.
Maas, J. 1999. Trace mineral requirements of cattle and poultry. In: Proc. Calif. Anim. Nutr. Conf., Fresno, May 5-6, pp. 39-48.
Mahan, D.C. and N.A. Parret. 1996. Evaluating the efficiency of selenium-enriched yeast and sodium selenite on tissue selenium retention and serum glutathione peroxidase activity in grower and finisher swine. J. Anim. Sci. 74:2967-2974.
McDaniel, G.R., J. Brake and M.K. Eckman. 1981. Factors affecting broiler breeder performance. 1. Relationship of daily feed intake level to reproductive performance of pullets. Poult. Sci. 60:307-312.
McDowell, L.R. 2003. Minerals in Animal and Human Nutrition. 2nd ed., Elsevier Science B.V., Amsterdam, pp 644.
Meuer, H.J. and R. Baumann. 1988. Oxygen pressure in intra and extraembryonic blood vessels of early chick embryos. Respir. Physiol. 71:331-342.
Noble, R.C. and M. Cocchi. 1990. Lipid metabolism and the neonatal chicken. Prog. Lipid Res. 29:107-
Pan, E.A., F. Rutz, N.J.L. Dionello, M.A. Anciuti and R.R. da Silva. 2004. Performance of brown egg layers fed diets containing organic selenium (Sel-Plex®). In: Nutritional Biotechnology in the Feed and Food Industries, Proceedings of the 20th Annual Symposium (Suppl. 1: Abstracts of posters presented). Lexington, Ky, May 24-26, p. 18.
Pappas, A.C., R.M. McDevitt, P.F. Surai, T. Acamovic and N.H.C. Sparks. 2004. The effects of selenium and PUFA supplementation in the diet of young broiler breeders on the incorporation of selenium in the egg and in the tissues of day old broiler chick. Br. Poult. Sci. S26-27(Suppl.).
Paton, N.D., A.H. Cantor, A.J. Pescatore, M.J. Ford, C.A. Smith. 2002. The effect of dietary selenium source and level on the uptake of selenium by developing chick embryos. Poult. Sci. 81:1548-1554.
Pedrieri, G. and E. Cinti. 2003. New metal chelates for animal nutrition. www.feedinfo.com (dated 20 June 2003; accessed 19 September 2003), 4 pages.
Pinchasov, Y. 1991. Relation between the weight of hatching eggs and subsequent early performance of broiler chicks. Br. Poult. Sci. 32:109-115.
Rahn, H. and C.V. Paganelli. 1991. Energy budget and gas exchange of avian eggs. In: Avian Incubation (S.G. Tullet, ed). Buttersworth and Heinemann, London, UK, pp. 175-193.
Rebel, J.M.J., T.P. van Dam, B. Zekarias, F.R. M. Balk, J. Post, A. Flores Minambres and A.A.H. M. ter Huurne. 2004. Vitamin and trace mineral content in feed of breeders and their progeny: effects of growth, feed conversion and severity of malabsorption syndrome of broilers. Br. Poult. Sci. 45:201-209.
Reddy, R.P. and M. Sadjadi. 1990. Selection for growth and semen traits in the poultry industry: what can we expect in the future. In: Control of Fertility in Domestic Species (J.P. Brillard, ed). INRA editions, Vol. 54, pp. 47-59.
Renema, R.A. 2004. Reproductive responses to Sel- Plex® organic selenium in male and female broiler breeders: impact on production traits and hatchability. In: Nutritional Biotechnology in the Feed and Food Industries, Proceedings of Alltech’s 20th Annual Symposium (T.P. Lyons and K.A. Jacques, eds). Nottingham University Press, UK, pp 81-91.
Renema, R.A. and F.E. Robinson. 2004. Defining normal: comparison of feed restriction and full feeding of female broiler breeders. World’s Poult. Sci. 60:508-522.
Romanoff, A.L. 1960. The Avian Embryo: Structure and Functional Development. MacMillan, New York, NY.
Rutz, F., E.A. Pan, G.B. Xavier and M.A. Anciuti. 2003. Meeting selenium demands of modern poultry:
responses to Sel-Plex® organic selenium in broiler and breeder diets. In: Nutritional Biotechnology in the Feed and Food Industries, Proceedings of Alltech’s 19th Annual Symposium (T.P. Lyons and K.A. Jacques, eds). Nottingham University Press, UK, pp. 165-185.
Sandoz, D., E. Ulrich and E. Brard. 1971. Etude des ultraestructures du magnum des oiseaux. Evolution au cours du cycle de ponte chez la poule Gallus domesticus. J. de Microscopie 11:371-400.
Shanawany, M.M. 1987. Hatching weight in relation to egg weight in domestic birds. World’s Poult. Sci. J. 43:107-115.
Surai, P.F. 2000. Effect of selenium and vitamin E content of the maternal diet on the antioxidant system of the yolk and the developing chick. Br. Poult. Sci. 41:235-243.
Surai, P.F. 2002. Natural Antioxidants in Avian Nutrition and Reproduction. 1st ed., Nottingham University Press, UK.
Surai, P.F. and N.H.C. Sparks. 2000. Effect of the selenium content of the maternal diet on the antioxidant system of the yolk. Department of Biochemistry and Nutrition, Scottish Agricultural College Auchincruive, Ayr, KA65HN, Scotland, UK, Br. Soc. Anim. Sci.
Suso, F.A. and H.M. Edwards, Jr. 1972. Binding of EDTA, histidine and acetylsalicylic acid to zincprotein complex in intestinal content, intestinal mucosa and blood plasma. Nature 236:230-232.
Tona, K., O. Onagbesan, B. de Ketelaere, E. Decuypere and V. Bruggeman. 2004. Effects of age of broiler breeders and egg storage on egg quality, hatchability, chick quality, chick weight and chick posthatch growth to 42 days. J. Appl. Poult. Res. 13:10-18.
Vander, A.J., J.H. Sherman and D.S. Luciano. 1990. Human Physiology. 5th ed., McGraw-Hill Publishing Co., New York, NY.
Vieira, S.L. and E.T. Moran. 1998. Eggs and chicks from broiler breeders of extremely different age. J. Appl. Poult. Res. 7:372-376.
Wakebe, M. 1998. Organic selenium and egg freshness. Patent No. 10-23864. Feed for meat chicken and feed for laying hens. Japanese Patent Office, Application Heisei 8-179629. Published Jan 27.
Wilson, H.R. 1997. Effects of maternal nutrition on hatchability. Poult. Sci. 76:134-143.
Wishart, G.J. and H.J. Staines. 1999. Measuring sperm:egg interaction to assess breeding efficiency in chickens in chickens and turkeys. Poult. Sci. 87:412- 466.
Xavier, G.B., F. Rutz, N.J.L. Dionello, A.D. Duarte, F.M. Goncalves and N.H.F. Zauk and C. L. G. Ribeiro. 2004. Performance of layers fed diets containing organic selenium, zinc and manganese, during a second cycle of production. In: Nutritional Biotechnology in the Feed and Food Industries, Proceedings of the 20th Annual Symposium (Suppl. 1: Abstracts of posters presented). Lexington, Ky, May 24-26, p. 19.
Authors: FERNANDO RUTZ, MARCOS ANTONIO ANCIUTI, JOSE LUIZ RECH and EDUARDO GONÇALVES XAVIER
Departamento de Zootecnia, Universidade Federal de Pelotas, Brazil