"NUTRITIONAL FORMULATION FOR POST HATCHED BIRDS"

FIELD OF APPLICATION

The present descriptive report refers to a "NUTRITIONAL FORMULATION FOR POST HATCHED BIRDS", destined preferentially, although not exclusively, to broiler-chickens raised for meat production. It contains a differentiated nutritional characteristic and permitting an efficacious gain in bodyweight. However the feed is also applied to day-old-pullets raised for egg production (laying hens and breeder hens).

Additionally it can bring significant positive impact on the immune system and internal organs development improving the chicks survival rates and maximizing their overall performance.

BASES OF INVENTION

The genetic progress obtained during these last decades by rigorous selection programmes is making the birds more and more productive. Allied to this progress the increasing knowledge on nutrition, health, placement facilities and equipments, plus better handling, efficient administration and economical control of costs, has resulted in a great evolution in performance of the contemporary birds as can be showed in table 1.

Table 1 - Evolution of the performance of broiler-chickens and of the consumption per capita of chicken meat.

Parameter ϊ950 ϊ970 ϊ990 2004 ^{i) 2004 ^(JJ

Live weight (g) ^ 1409 1681 2045 1882 2272 Slaughter age (days) ⁽¹⁾ 70 56 39 35 42 Feed conversion ⁽ⁱ ^ 3 2,2 1,9 1,59 1,72 Feed efficiency (%) 33,33 45,45 52,63 62,89 58,13 Consume per capita _ 2,3 14,2 33,8 33,8

(kg/hab.year) ⁽²⁾

Adapted: ^UJ - Desouzart (1994); ^w - UBA (2002) e EMBRAPA - CNPSA (2004); ⁽ⁱ⁾ - World Poultry (2004).

Application of advanced techniques in sanity and ambience have enabled the chicken raise in higher densities, and reductions in the negative impact of diseases on the raising. Sophisticated systems of control of lighting, temperature and humidity in the sheds, as well as the increase of the immunity of the birds, through more effective vaccines have allowed the expression of the earnings provided by the genetics. Use of modern feeders decreases feed wastage and provides a greater stimulus to consumption.

Procedures of specific handling for the several phases of the raising, since the placement of the chickens to the bird catches for slaughter, as well as its transportation for the slaughterhouse have become routine in the poultry industry, bringing up much more gains on productivity with the reduction of bird condemnations in the slaughterhouse.

Finally, the professionalism of the agribusiness, with implantation of specific departments for purchasing, sales, accounting and financial administration, has provided to the larger companies greater efficiency. The nutrition is taking advantage of all this once the raw material, being purchased could be a flock better in price and quality. Good raw materials allow the nutritionists to have a more precise evaluation of feed ingredients which, in turn, would permit the fine tuning in the nutritional formulation that could result in a bigger return over investment.

It can be observed in Table 1 above that in 1950, the weight of birds was 1.4 kg, after 70 days of age, while in 2004, average weight was 34% higher in half the time, with a feed efficiency 89% higher, which implies that while in 1950 30 kg of feed was used to produce 1.0 kg of birds by live weight, in 2004 this consumption fell to 1.59 kg, representing a reduction of 47% in consumption of live chicken, making a significant impact and favoring the cost of production, which explains the increase in consumption per capita of chicken in Brazil from 2.3 kg per inhabitant per year in 1970, to 33.8 kg per inhabitant per year in 2004 (EMBRAPA-CNPSA). From the increase in feed efficiency, it can be presumed that there is a marked reduction in production of wastes per chicken produced.

Thus in the last five decades, the age of slaughter has been reduced by approximately one day per year. In 1950, the first week of life corresponded to 10% of its lifetime, as slaughtering was at 70 days of age, and when the birds would be approximately 1.4 kg. Today, the first week of life corresponds to approximately 20% of the birds' lifetime, considering that it is slaughtered at 35 days at 1.88 kg in weight, or 17%, considering the

birds to be slaughtered at 42 days at 227 kg weight, and consumption of feed in this phase represents only 5% to 10% of total volume.

A recent survey of the main poultry companies in the sector, as well as a survey by Nascimento et al. (2005), Burin (2004) and Penne (2005), cited by Nascimento (2005), found that use of pre-initial feeds by broiler chickens is currently common practice adopted by most Brazilian companies, in contrast to previous observations published at conferences in Brazil by Nir (1998), Penz and Vieira (1998) , Toledo et al. (2001), Vieira

(2004) and Araύjo (2004), that although there was unequivocal proof of the advantages in using pre-initial diets for broiler chickens, (1 to 7 days), there was still residence on the part of big companies mainly due to logistical problems involved in manufacturing and distributing one more diet at the poultry houses.

Araύjo (2004) showed that feeding in the first hours of life not only provides greater speed in growth and feed efficiency, but also involves resistance to the many pathogens, as well as greater uniformity in the flock. Chicks of 4Og weight at placement reach, in 7 days, weights of 160 to 20Og, with an average of 180g; that is, in the first week the chick should at least double its initial weight, and can quintuplicate it, so long as special nutritional conditions, environment, management, sanitation and genetics are assured.

Experiments have demonstrated that gains of 10 grams in weight at 7 days result in more than 50 to 70 grams in weight at 42 days of age (Navarro, 2004).

The optimization of this growth in the post-hatching period is dependent upon a complete chain of good practices since the breeders rearing conditions, which include their age, nutritional regime, overall environment (temperature, humidity, lighting, etc.) health, proper handling and genetics. The quality of the chickens, themselves, is dependent also on the egg manipulation, storage temperature, sanitizing procedures, hatchery machines management and proper temperature and humidity controls plus high hygiene status, well trained personal and a efficacious vaccination program, among others. Transportation time, overall ambient of the vehicle and the time since hatching up to destination and placement and proper handling by well trained personal, etc, also affect the bird's performance (Kidd, 2002).

The data in table 2 show significant differences in the bird's bodyweights at birth, arising of the effects of different conditions pre-hatching, they provide expressive differences in the weight at 21 and 42 days.

Table 2 - Effect of the weight of chickens at birth on the weight at 21 and 42 age days of broiler chickens.

Weight 1 day (g) Weight 21 days (g) Weight 42 days (g)

34,4 c 552 c 1872 b

38,2 be 579 be 1902 b

42,0 b 606 ab 1996 a

45,7 a 627 a 2039 a

P=O ₅ OOOl P=0,0001 P=0,0001

Adapted of the University of Alberta (Canada) data, cited by Tzschentke, 2002.

Nutrition of the chick immediately after hatching has become an important preoccupation in current poultry raising. Feeding right after birth of the chick, as well as promoting greater speed in growth and feed efficiency, also favors development of the immunological system, making the animals more resistant to pathogenic organisms.

Normally, the chicks fast for long periods before being housed in the production sheds, which definitely hinders development of the birds. On the other hand, birds that hatch sooner remain in the hatcheries for several hours until hatching is complete in the entire hatching room. After removal from the hatcheries, the chicks must be vaccinated, sexed and placed in transport boxes, which further increases the time in the hatchery and stress in the period.

This pre-placement time can be of more than 40 hours, including the time for transport from the incubator to the shed. In this transport, the chicks suffer dehydration, resulting in loss of weight, growth retardation, lessened resistance to infections and increased mortality. This pre-placement fast delays development of the birds' intestinal mucous, retarding absorption of nutrients.

Corporal development of chicks can be advanced by early access to the diet. The benefits of this early consumption are more pronounced in live weight in the first week and this advantage is maintained to slaughter.

Post-hatching factors and development of broiler chickens:

In order to understand clearly the mechanisms by which it is possible to manipulate nutrition and management of chicks, with the objective of improving performance, it is necessary to first understand how the birds' body develops, that is, the alometric growth of its vital organs; the standards of secretion of digestive enzymes, changes in metabolism, and other things. Changes in the gastro-intestinal tract after hatching:

Digestive enzymes are already active in the embryo, as also the mechanisms for absorbing nutrients by the intestine. Extracellular enzymes that are secreted by the endoderm of the vitelline sac act on the substrate, enabling absorption of the products of digestion, including macromolecules. The vitelline content at hatching represents approximately 20% of the weight of the chick It is made up of approximately 46% water, 20% protein and 34% of lipids. In other words, a chick of 4Og weight has approximately 8g of vitelline content with 2.72g of lipids and 1.6g of proteins (Sklan & Noy, 2000). The vitelline content at the moment of hatching comes from remaining portions of yolk and albumin, with this last flowing to the vitelline sac at the end of the second week of incubation, when the sero-amniotic connection ruptures, and it is almost all "ingested" by the embryo; however, part of the albumin moves to within the vitelline sac increasing its protein content (Vieira, 2004). The embryo absorbs the vitelline content simultaneously through two main pathways:

In one of them, the vitelline content is transferred, in the form of lipoproteins, directly to embryonic circulation, through a process of phagocytes or endocytosis.

In the other pathway, the vitelline content is transferred to the large intestine, where, by antiperistaltic mechanisms of the mucous, it is taken to the region of close to the large intestine, undergoing action by enzymes such as lipase, allowing its digestion and consequent absorption by the intestinal mucosa (Noy & Sklan, 1998). It should be pointed out that the vitelline sac membrane can be seen as an extension of the embryo intestine, and is subject to the contractions and movements of the same (Vieira, 2004).

The embryo intestine contains enzymes that can digest proteins such as cheimotripsine and carboxypeptidase A, and trypsin is also present, however with its activity inhibited by the ovomucoid fraction of the albumen, whose evolutionary significance is related to inhibition of the activities of other protolytic enzymes activated by tripsin, which

would result in degradation of IgA and IgG, present in the albumen and important in the passive immunity of the chick in the first days following hatching, a period in which its immunological system is immature (Vieira, 2004).

Around 20% of the proteins in the remains of the vitelline sac are immunoglobulins of high diversity of mother origin, which would not be able to be produced by the embryo. The use of this protein fraction, as a source of amino acids, for metabolism of the embryo, would deprive the newly born of this important mechanism of immunological defense in the first week of life. Thus, the use of this protein fraction in cases of fasting should not be interpreted as a normal metabolic pathway, but rather as a survival mechanism for the chick, where this deviation of function can cause harm to the chicks potential development.

The remaining 80% of protein of the vitelline remains are made up of seric proteins, present in the hen at the moment of forming the yolk, they can contain antigens to which the chick can be exposes in the pre-hatching environment. During the first 48 hours, use of the vitelline sac, through a circulatory system, remains functional, however, after this period, transference begins to reduce by the obstruction of the vitelline peduncle by lymphoid cells, which is completed after approximately 4 days from hatching. (Noy & Sklan, 1998).

Close to hatching, between the 19th and the 20th day, the residual vitelline sac internalizes in the abdominal cavity, and at birth, the intestine contains a viscous material coming from the yolk (Noy & Sklan, 1998).

Right after hatching, the chicks react strongly with the environment, attempting to peck and eat particles, with changes in the morpho-physiological structure of the intestinal tract, as with the ingestion of food, maturation of the digestive organs is accelerated, as well as the reserves of the vitelline sac, due possibly an increase in intensity of ant-iperistaltic movements in the intestine.

Experiments carried out with chicks revealed that removal of the vitelline sac reduces the performance of the birds, demonstrating the importance of the vitelline sac reserve for chicks right after hatching (Edwards et al. 1962). In the period right after hatching, the weight of the chick's increases with greater velocity than its body weight as a whole. This process in the development of the chicks intestine reaches a maximum peak at around 6 to 8 days; however, other organs of the

digestive system such as the pancreas and gizzard do not show the same rhythm of growth as relative weight.

Studies on the development of the area and the height of intestinal mucous velocity show significant differences. Studies in the development and the area of the velocities of the intestinal mucous show that there are significant differences in the different portions of chicken intestines. The greatest velocity in growth relative to area and height of intestinal velocities reaches a plateau in 6 to 8 days, in the duodenum portion, and at 10 days, at portions of the jejunum and the ileum (Noy & Sklan, 1998).

Secretion of pancreatic enzymes increases with the age of the birds and with consumption of food; however, when the secretion of enzymes per gram of food ingested is calculated, there are no significant changes for the enzymes amylase, trypsin and lypase, between 4 and 18 days. Similar results were obtained for secretions of bile salts and free fatty acid in the duodenum. However, the quantity of nitrogen secreted in the intestine per gram of food ingested is small during hatching and increases with age (Noy & Sklan, 1998, 2000). Feeding chicks stimulates the secretion of the pancreatic enzymes amylase, trypsin and lipase, and the use of adequate levels of sodium is essential for mechanisms of absorbing nutrients, especially glucose and amino acids by the intestinal mucous. (Noy & Sklan, 2000)

The activities of the enzymes maltase, sucrose, alfa-glutamiltransferase and alkaline phosphates released by the membrane cells of the "borda em escova" (microvilosities of the intestinal mucous) increases with age, and this increase of activity is correlated with the beginning of ingestion of food (Noy & Sklan, 1998).

Mahagna & Nir (1996) observed that activity of the dissaccaridases of the intestinal mucous increases with the age of chicks, with a reduction seen in activity of dissacaridases being observed during the period of 0 to 4 days of age, which could be caused by dilution of chemicals in the intestine, and by the process of maturation and turnover of enterocytes.

Considering that maltose is the main product in digestion of starch in diets based on corn and soybeans, Shapiro et al. (1997) they concluded that production of dissacaridases in the intestinal mucous of broiler chicks could limit performance. These authors made tests using broiler chicks fed with diets containing 20% glucose or 20% maltose. Supplementation of the maltose significantly reduced the weight off the chicks when compared with birds fed on diets containing 20% of glucose (Table 3).

Table 3 - Effect of addition of glucose and maltose in the diet on the performance of broiler chickens.

Age Live Weight (g) Control +20% glucose +20% maltose

7 days 126 136 123

14 days 330ab 355a 310b

21 days 677ab TlOa 621b

Shapiro et al. (1997).

The absorptive capacity of the intestinal mucous immediately after hatching has also been the subject of studies. Experiments in vitro and in situ demonstrarate that in the chick immediately after hatching, absorption of methionine and glucose has limitations, increasing with age, especially in the duodenum. On the dother hand, the absorptive capacity of fatty acids shows no changes with age, and is high immediately after hatching. Sklan (2003), suggests that, due to the fact that in the embryo there are active mechanisms for digestion and absorption of fats from the vitelline sacsin post-hatching, these mechanisms need few adaptations to the new substrate in which they will act. On the other hand, lower absorption of glucose is due to the fact that in the vitelline content, there is almost no carbohydrate. There is also a low concentration of sodium in the intestine, which is required for good functioning of Na-glucose cotransporters, and inhibition of absorption of hydrophylic compounds such as glucose, due to the hydrophobic quality of the vitelline content (Richards, 1997).

Previous studies suggest the chicks have a sub-optimum absorption of lipids from the diet (Krogdahl, 1985). However, determination of absorption of lipids from metabolism tests is complicated by the presence of residual contend of vitelline in the excretia, which causes an underestimation of digestibility of fats. However, PoUn & Hussein (1982), found that supplementing the diet with bile salts improves the digestibility of fats in chicks of 7 days of age. Krogdahl (1985) observed that secretion of these bile increases up to day 21 after hatching, when it then declines. Noy & Sklan (1995) observed increases of 8 to 10 times in the secretion of bile components, such as bile salts and fatty acid between 4 and 21 days of age, with the greater part of fatty acid secreted in the bile in the form of phospholipids. Bile secretions are low after hatching and increase with age and can limit

absorption of fats, as in some studies, it was observed that addition of cholic acid improves digestion of fats in chicks (Serafim & Nesheim, 1967; Gomez & Polin, 1976; Polin et al. 1980).

Composition of the lipid fraction of the yolk is 65% of triacilglycerides, 25% of phospholipids and 10% of esthers of cholesterol, the last two being respectively important in formation of micelas of fat in the intestine, aiding in digestion of the same, and precursors of the formation of bile salts and hormones.

Recent studies by Sklan (2003) demonstrate in the chick, after hatching, that the absorption of glucose in the circulation increases with age, while the rate of use of the same for producing energy remains constant. In contract, the rate of absorption of fats is high at hatching, remaining unaltered with age, while use of fatty acids for production of energy decreases with age.

Appearance in the circulation of lipids originating from the vitelline sac or from the intestine are high and similar, showing an initial increase in its rate of utilization and later, a constant decline.

Distribution of marked lipids in the classes of lipids and fractions of lipoproteins was examined, there being few alterations in these standards with age. However, marked lipids in the vitelline residue were preferentially found in the LDL fraction, lipoproteins of low density, which corroborates previous observations that a significant part of the vitelline content is transported directly by the circulation. Lipoproteins are synthesized in the embryo, in part, in the endodermal epithelium of the yolk, and in part from maternal VLDL present in the yolk that is remodeled in the endodermal epithelial cells. Concentration of LDL and HDL decreases rapidly in chicks up to 5 days after hatching. Thus we can conclude that immediately after hatching, the lipoproteins originating from the yolk or its endoderm are responsible for most of the transport of lipids in circulation. However, these lipoproteins are metabolized immediately after hatching and thus the concentration of LDL and HDL fall, affecting the turnover of plasmic lipids, and consequently their use for production of energy. Transport of lipids recently absorbed in the large intestine occurs normally in the liver, where they are re-exported in new particles of lipoprotein synthesized at this location.

In summary, in the chick, at the moment immediately after hatching, lipids from the vitelline content residue are transported by the circulation in the form of lipoproteins,

and are later oxidized for production of energy, while external supply of glucose is still low.

After a few days, the lipids from the vitelline content residue and the lipoprotein for their transport are reduced, causing a fall in the use of fatty acids to produce energy. It is possible that the production of lipoproteins can limit, therefore, the use of fats in the diet, seeing that chicks do not seem to respond to high levels of fat in pre-initial diets (Noy & Sklan, 2001 and

2002). Furthermore, external supply of glucose becomes the main energy source for the chick.

Thus, the change in the metabolism of dependance on the use of lipids from vitelline residues as the main energy source to dependence on the use of glucose is accelerated by the lowering in the concentration of lipoproteins in circulation. The speed with which food passes through the gastro-intestinal tract is important, as it determines the time that the same will be exposed to digestive enzymes and the mechanisms of absorption of nutrients by the mucous. Immediately after hatching, and with ingesting of food, the rate of passage increases significantly (Vergara et al. 1989), then decreasing between the 4th and 10th day after hatching by approximately 30%, although consumption of food increases 3 fold. The decrease observed in the rate of passage is more accentuated in the duodenum. Between the 10th and the 21st days, no changes are observed in the rate of passage, although consumption of food continues to increase (Noy & Sklan, 1995). However, in previous studies, by Sklan et al. (1975) rates of passage were observed in the large intestine between the 42nd and the 49th day (a rate of 67 minutes by the large intestine) which indicates the possibility of there being a reduction between 21st and the 49th day.

Studies by Noy & Sklan (1995) report that at 4 days, for chicks receiving a diet with 6% fat, mainly unsaturated, the coefficient of digestibility of fat was above 85% at 4 days, with a slight increase up 21 days, indicating that sufficient quantities of lipase and bile salts were available at 4 days of age. The same was absorbed with digestion of carbohydrates, with values of digestibility also around 85%. Digestibility of protein increased from 78% to 80%, from the 4th and 7th day respectively to 90% on the 21st day.

The reserves of the vitelline sac are almost exhausted by the 3rd or 4th day after hatching. However they are sufficient to provide only 50% of the energy and 43% of the protein on the 1st day of life, while the use of fat is faster than protein (Murakami et al. 1988). Thus, it can be seen that the main functions of the vitelline reserves are to aid in maintenance of life of the birds during its transition to independence, during the first moments of learning to find its own food.

In the evolution process, nature made the egg a food rich in energy, and essential amino acids, vitamins and other nutritionally important components. In humans, absorption of nutrients of a boild egg reaches 95%. Its protein is considered standard protein with a value of 100 in the NPU index (net protein utilization), while protein from fish has an average value of 85, cows' milk is 75, rice is 57, wheat is 52 and beans 47. Some digestive enzyme inhibitors are present in eggs and their function will be discussed further on, but they mostly make their digestion difficult by wild predators, and to allow transference of immunoglobins (protein) to the chick. Knowing the nutrient composition of eggs allows us to make inferences and plan possible manipulations of the diet of chicks with a view to improving their performance.

Incubation of an egg is approximately 21 days, during which period the embryo has as its only source of food, the constituents of the egg. The presence of carbohydrates is almost nil; however, some tissues, such as the brain, red blood cells, renal medulla and others, can use glucose as a source of energy, except in certain special conditions where cetone bodies could be used. Thus glyconeogenesis, which is the formation of glucose from other compounds, such as amino acids for example, is a highly active process in the embryo.

The availability of glucose is especially important at the immediate moment preceding hatching, when there is the transition from corion-alantic respiration to complete dependence on pulmonary respiration (White, 1974, cited by Vieira, 2004).

With the beginning of pulmonary respiration and hatching, demand for glucose increases in the organism. The main organ producing glucose, by glyconeogenisis, is the liver, which responds for 90% of the total. The reserves of glucose run out quickly if the gluconeogenesis process is not active. Long periods of fasting, in which carbohydrates are not available for the chicks, increase the demand for gliconeogenese of proteic origin, which is the formation of glucose using aminoacids as substrate, and the use of fat as a source of energy, with respective increase in the plasmatic concentration of cetone bodies, as well as reduction in the quantity of metabolic water, which is important for rehydration of the birds (Best, 1996 and Hammond, 1944, cited by Vieira, 2004). Placement of chicks with food and water should be as fact as possible, respecting the minimum necessary time of the birth installation. Loss of weight and dehydration caused by fasting, even at a minimum, can cause an increase in mortality,

retardation of development of intestinal mucous, thus causing less efficiency in digestion and absorption of nutrients. Old recommendations for management cited fasting as a practice for improving performance, as it favored a faster reabsorption of the residual vitelline sac. This practice proved to be inappropriate as it has been amply demonstrated that ingestion of external food accelerates use of the residual vitelline sac.

Normally, chicks arrive at the granges 24 to 36 hours after hatching, with this time spent in procedures for sexing, vaccination, transport and other things, while during this time, the chicks lose weight by use of the vitelline sac, by digestive and renal excretions, and by dehydration. Noy & Sklan (1997) found that increase in weight of chicks only occurs 36 to 48 hours after having access to a diet. The beginning of this growth can be anticipated by bringing this access forward. The benefit of this anticipation of consumption is most pronounced in weight gain at 7 and 10 days of age, while the advantage gained continues up to slaughter.

Noy & Sklan (2000) found that chicks not provided with exogenous food for 48 hours after hatching suffer diminished weight. However, during these 48 hours the weight of the intestine increases 60% in chicks with no food and 200% in chicks provided with food.

The energy for maintenance for chicks in the first 24 hours has been estimated at approximately 11 kcal (112 kcal . W 0.75). Assuming that all residual content of the vitelline sac released in the first 24 hours were used only as a source of energy with 100% efficiency, there would be only 9.4 kcal. Thus, without additional supplies of nutrients the chick would enter a negative energy balance and certainly lose weight (Dibner et al., 2005)

Specific diets have been developed for use in situations that were previously practically ignored by the scientific community. For example: nutrition in the egg and nutrition while still in the incubator, through special diets where the objective is to make the transition from pre-hatching and post-hatching more smooth, avoiding large "leaps" in metabolism changes; possible use of pro-biotics to assure the first colonization of the digestive tract; use of sodium salts to overcome the deficiencies of the yolk in this ingredient, and use of bile salts to enable better use of fats in the diet, have been tested and obtained promising results. In general, complete adaptation of the digestive tract and the metabolism of the chick to use a diet rich in carbohydrates and relatively poor in fats, in substitution of using the vitelline sac, lasts between 3 and 4 days. The digestive tract of the chick at the exact moment

of pre-hatching has available only the residual vitelline sac to effect processes of digestion and absorption. This substrate is rich in fats and proteins and almost absent of carbohydrates, and therefore has very high hydrophobia characteristics. Feeding at the moment of post- hatching, as it is rich in carbohydrates, as well as having low hydrophobia characteristics, requires digestive enzymes such as amilase, maltase and others. ANTECEDENTS OF TECHNIQUE

Some contributions are known for the state of the art in developing nutritional formulations for chick rations; patent document US 4556564 refers to a formulation of feed containing the component zeolita as an ingredient for improving resistance of the shell and generally desirable characteristics of eggs.

Patent document JP 2004329073 refers to an additive to feeds for broiler chickens that comes from brown algae, and varieties of camellia and angelica in which the formulation provides an improvement in growth and production on battery scale without causing stress to the confined birds. Patent document NZ 536677 refers to an additive based on a dicarboxylic alpha amino acid that provides a bioavailability of minerals and amino acids to the animal without generating serious pollutants.

In all cases, the teachings of the state of the art associate feeding with fattening and increase in weight, while feeding should be associated to development of the animal, with emphasis on healthy meat and desired nutritional contents.

OBJECTIVE OF INVENTION

The objective of the present intervention is to enable nutritional formulation of a feed with up to 18% humidity, that brings improved results in zoological-technical performance of poultry during their production cycle, advancing maturity of the immune system, making the birds more resistant to illnesses, improving response to vaccines and increasing survival; better protection of the intestinal mucous, and precocious development of organs of the digestive and cardiorespiratory system, to allow the birds to sustain accelerated development. SUMMARY OF INVENTION The present patent is for a nutritional composition for a feed with up to 18% humidity and a period and way for supplying it, that has the results of an improvement in results of zoological-technical performance for birds during their productive cycle, advancing

maturation of the immune system making the birds more resistant to illnesses and improving response to vaccines and increasing survival; better protection of the intestinal mucous, and precocious development of organs of the digestive and cardio-respiratory system, to allow the birds to sustain accelerated development. The period recommended for supplying the product is between hatching and placement in granges, a period henceforth called post- hatching. The feed can be provided on the floor of the boxes for transporting the chicks or on the floor of the trays of hatcheries in the incubator room. Remains of feed in the transport boxes, after placement, can be offered to the birds on the floor on paper. This feed may or may not contain a probiotic additive in its composition with the purpose of colonizing the digestive tract with strains of specific microorganisms that are beneficial to the birds. In another modality of the patent, the probiotic additive may be accompanied by a prebiotic additive, with the purpose of serving as a substrate for the strains of microorganisms mentioned previously, accelerating multiplication of the same in the digestive tract.

In another modality of the patent, the feed may contain coccidiostatic or anticoccidian substances, such as ionphores, in order to reduce proliferation of Eimerias in the digestive tract of the birds. The anticoccidian or coccidiostatic agents may or not be associated with antibiotic agents that promote growth, that are gram negative and/or gram positive.

As na option, one or more of the following products may be added to the product ^• . Probiotics, Prebiotics, Anticoccidians, coccidiostatics, antibiotics, Cisteamine, enzymes, food colorings, adsorbents, mannanoligosaccarides, imunoglobulins, transferrina, lecithin, bile salts, emulsifiers, glutamine, glycine, serine, lysine, methionine, treonine, arginine, tryptophane, valine, leucine, isoleucine, nucleotides, boron, strontium, molybdenum, chromium, carnitine, fatty acids, Omega 3 (DHA and EPA), amides, dextrose, acidifying additive, cholesterol etc. DESCRIPTION OF PROFESSED MODALITY

The present invention is described in terms of its preferred modality which is the result of a study for maximizing animal development.

Maximization of muscle development by means of post -hatching diets.

Foπnation of skeletal muscle fibers is taken to be finalized at hatching for birds, it not being possible, under normal conditions, and after this moment, for there to be an increase in the number of muscle fibers by mitosis of miofibrils. However, a significant increase in muscle DNA can be seen during post-hatching growth in birds, with this process being essential for muscular hypertrophy. The increase in muscle DNA takes place due to the activity of satellite cells, myogenic precursors present in the skeletal muscles, which begin their development during the last embryonic phase, and can proliferate, differentiate and join to the existing fibers, or fuse with others, forming new fibers. At the moment of hatching, there is a high number of satellit cells that, with the beginning of growth, decrease, remaining at rest, and being reactivated only in cases of muscular damage (Schultz et al., 1978 and Mauro, 1961, cited by Araύjo, 2004).

At the moment immediately after hatching, there are still a great number of these cells, with their nuclei responding for all those incorporated in normal muscle fibers after birth. Each DNA of multinucleated cells controls a volume of cytoplasm in the immediate vicinity through RNAm. Thus the rate of muscular growth (hypertrophy) depends on the aggregation of new muscle fiber nuclei. Making nutrients available after hatching of chicks is a primordial condition for stimulation and proliferation of satellite cells and their incorporation into muscle fibers for maximum muscle growth (Moss & Leblond, 1971, Halevy et al, 2000. cited by Vieira, 2004, Moore et al. 2005). Consequently, periods of prolonged fasting that cause the "death" of muscle cells in birds (apoptosis), would bring irreparable damage to the muscle development of birds, affecting the quality of the body at slaughter. Post-hatching diets and development of the immune system.

Genetic selection of birds, with the objective of increasing the velocity of growth, has had a negative impact on immunocompetence. There has been a reduction in the capacity for production of antibodies, making them less resistant to pathogens Bigot et al. 2001, cited by Araύjo, 2004, as in Table 4.

Table 4 - Antibodies of two lines of chickens according to their genetic potential.

Adapted from Cheema et al (2003), cited by Araύjo (2004)

On the other hand there was na increase in inflammatory responses caused by cells with na increase in production of lymphocytes and macrophages, consuming nutrients and causing a reduction in consumption of feed, as can be seen in Table 5,

Table S - Proliferation of lymphocites and the activity of macrophages in two lines of broiler chickens according to their genetic potential.

Adapted from Cheema et al (2003), cited by Araύjo (2004)

With genetic selection there is also a diminution of relative weight of lymph organs, although absolute weight of these increased in function of the increase in the average weight of the birds, as shown in Table 6.

Table 6 -Body weight and its relation to lymph organs of two lineages of broiler chickens according to genetic potential.

Adapted from Cheema et al (2003), cited by Araύjo (2004)

The capacity of birds to deal with stress situations has changed, especially in modulated responses or those regulated by hormones. Birds have been raised in increasingly high densities and submitted to greater microbial pressures, despite the greater sanitation of modern granges.

The immunological system of birds begins development in the embryonic phase and is partially developed at the moment of hatching. The main organs of the immune system (thymus and bursa) are present in the active lymphatic system. Migration of lymphocytes to the thymus takes place in waves beginning from the sixth day of incubation.

They pass the thymus and remain in peripheric tissues.

Proliferation of lymphocytes in the bursa occurs between the tenth and fifteenth day of the embryonic phase. These cells are differentiated B cells and can express IgM only at the moment of hatching. Secondary immunological organs such as the spleen, cecal tonsils, Meckel's diverticle, the Harder gland and lymphoid tissues diffuse in the intestine and the respiratory system, are immature at hatching. At the moment of hatching, there are B cells in the tonsils B, but they express only IgM. In the coating itself and the intestinal epithelium, as in other secondary organs of the immune system, there are T cells, but immature, without cytotoxic capacities or for combating antigens until a few days after hatching. The ability to generate a secondary immune response, indicated by the presence of germinal centers, or, for IgG and IgA circulation, begins to manifest only between 1 and 4 weeks after hatching.

Removal of the spleen in newly hatched does not cause severe loss to responses by T cells, indicating a high degree of development during embryo genesis. On the other hand, removal of the Bursa on the 18th day of incubation can result in total loss of IgG and IgA, resulting in only primary responses of IgM of very limited diversity as the only capacity of humoral immunity.

In chicks at the moment of hatching, the predominant imunoglobulin in the bursa is IgM. The lymphocytes carriers of IgM are the precursors of the cells producers of IgA and IgG. The IgG present in the Bursa at the moment of hatching is in the connective interfolicular tissue, specifically in the blood vessels, and originates from the maternal matter in the vitelline. Thus the chick, when it is born, does not have the capacity to produce IgG, and is totally dependant on maternal antibodies for humoral protection. IgA is not present in the Bursa, as the chick is incapable of producing it. Similar observations can be made in cecal tonsils and other secondary organs of the immune system. Thus, it can be concluded that the immune system of chicks at hatching consists in production of IgM and stored maternal IgG. To deprive chicks of feed immediately after hatching causes a more accentuated reduction in the weight of the Bursa than the loss of body mass in itself. Providing feed on the 3rd day after hatching does not correct this loss, and the Bursa will continue smaller at least up to 21 days of age. Loss of relative weight of the Bursa associated with fasts can be related to the increase in glycocorticoids which are associated to involution of lymphoid organs or even the low availability of substrates or the presence of antigens in the organism.

The development of organs in chicks after hatching has suffered strong influences from the process of selection. High priority should be given to the digestive, circulatory and respiratory systems, while it can be supposed that lack of nutrients for development of organs and tissues related to immune responses have been impaired.

Development of secondary organs of the immune system, such as the spleen, cecal tonsils and the Harder gland are clearly dependant on the Bursa and the Thymus, but exposition to antigens is also na important factor. Chicks free of pathogens (Germ-Free) show small cecal tonsils, without germinative centers, and only after 4 weeks, do some germinative centers appear.

Ingesting food in itself is considered exposition to antigens, as well as being a source of nutrients and avoidance of depressing the immune system, as occurrs when there is fasting. Dibner et al. (1998) demonstrated that chicks fed with a hydrated nutritional supplement had a high proliferation of lymphocytes in the Bursa 3 days after hatching. On the contrary, chicks in fasting had na absence of lymphocytes, demonstrating that residual control of the vitelline sac present in the chick after hatching is not a substitute for exogenous feeding.

Appearance of IgA (the last of the main isotypes of Imunoglobulins) is considered to be a sign that the humoral system is fully developed. The presence of this imunoglobulin in bile secretions in chicks that receive supplementary feeding of hydrated feed for three days after hatching increased more rapidly when compared with chicks subjected to 3 days of fast.

The presence of germinative centers is another sign of the imunocompetence of birds, hi these places there is the significant presence of T and B ₅ and antigen receiving cells, that are important in the development of the immune system, as for example those required in vaccination responses. The germinative centers in the cecal tonsils were already present in the eighth day after hatching in chicks that received supplementary feeding for three days, compared with chicks under fact, where the centers only appeared after 15 days.

In the same study, there was also a challenge of coccidiosis in the birds. Birds challenged by coccidiosis showed less gain in weight and in efficiency than birds that were not challenged. On the other hand, birds that underwent fasts for three days after hatching showed less gain in weight and nutritional efficiency than birds that received hydrated food supplements.

The same authors cite that there are three main ways in which neonatal feeding can influence birds immunity. First as a supplier of substrate to tissues, organs and specialized cells, etc. of the immune system, secondly by the immuno-modulating substances present in the food, such as specific fatty acids such as arachidic acid, docohexanoic acid (DHA) and arachidic acids (EPA), and finally, by being a vehicle for antigens that will stimulate appearance of specific isotypes of imunoglobulins.

Wang et al. (2004), by manipulating the concentrations of specific fatty acids in the diet of light hens, managed to influence the passive immunity (IgG) transmitted to eggs. Klasing (1998) affirms that deficiencies in specific micronutrients can be as damaging or more so than deficiencies of macronutrients in the development of the immune system. Vitamins A and D act, for example, on the process of differentiation in the precursor cells of the immunity system. Deficiency in vitamin A in the diet results in birds with lower resistance to infection challenges. The estimated requirements of vitamin A for maximum weight gain and for maximum immunocompetence for birds differs in about 10 to 20 times (Friedman & Sklan, 1997, cited by Araύjo, 2004).

Nutritional value of feed for chicks.

The effect of age of the birds on the nutritional value of feed has been the subject of many studies. These studies have shown that the value in tables of feed compositions are normally made with birds of 16 to 25 days or adult roosters, and are not appropriate for the diets of chicks.

For Nir (1998), the values for EM in feeds found in tables of compositions are well above those realized by chicks in the first week, especially for those feeds that provide an increase in intestinal viscosity. Table 7 shows the values for EM determined for chicks in the first week (NIR, 1998) and for birds of different ages; be they chicks, roosters or hens (Rostagno et al., 2000).

Table 7 - Amounts of EM (kcal/kg DM) of the feedstock based on the dry weight that was determined in chick in their first week and in birds within the range of 15 to 21 days of age.

Age 1st Week Varied

Com 3244 3870 ^"

Sorghum 3118 3681

Wheat 2811 3506

Soybean Powder 1124 2572

Source Nir (1998) Rostagno et al. (2000)

In a similar way, Batal & Parsons (2002) demonstrated that the age of the birds not only interfered in the amounts of metabolizable energy, but also in the apparent digestibility of various nutrients in the diet, as showed in table 8, having also been observed that, applying a linear regression and plateau, the value of EM increases to the 14th day and the digestibility of the lysine to the 10th day, after which they remain relatively constant.

Table 8 - Effect of the age of the broiler chick on the values of EM (kcal/kg) and the coefficients of apparent digestibility of lysine and threonine of different diets.

Age 1st Week Varied

Corn 3244 3870

Sorghum 3118 3681

Wheat 2811 3506

Soybean Powder 1124 2572

Source Nir (1998) Rostagno et al. (2000)

Type of diet Age (Days)

0-2 3-4 14 21

EM (Kcal/kg)

Corn + Soybean Powder 3021 3078 3171 3347 3340 AAs Crystalline 1 3653 3635 3773 3850 3871 Dextrose, casein 2 3796 3794 3785 3786 3816 Coefficient of Digestibility Applicable to Lysine (%)

Corn + Soybean Powder 76 74 84 86 88 AAs Crystalline 1 93 93 96 97 98 Dextrose, casein2 96 97 97 97 98

Coefficient of Digestibility Applicable to Threonine (%)

Corn + Soybean Powder 70 64 73 81 81

AAs Crystalline 1 92 89 91 93 93

Dextrose, casein2 95 94 94 94 97

(1) Amino acids (20.36%) and corn starch (57,33%); (2) Dextrose (63,4%), casein (20%) and amino acids (4,79%); Batal & Parsons (2002)

The energetic values and the digestibility of the amino acids of the dextrose- casein diet were not affected by the age of the chick; on the other hand, the values of EM of the diet with synthetic amino acids and starch increased with the age, a factor which shows the difficulty of the broiler chick in digesting starch (Table 9).

Table 9 - Effect of the age of the chick on the values of EM and the apparent digestibility of the nutrients in the diet of corn, soybean cake and oil.

Nutrients Age (days)

0-2 3-4 7 14 21

EM (kcal/kg) 2970 3085 3185 3429 3426

Starch (%) 93 93 97 99 99

Fat (%) 61 58 59 74 73

Lysine (%) 79 81 85 89 89

Metionine (%) 80 82 87 92 92

Threonine (%) 69 70 76 88 85

Regression and plateau for EM = 14 day; for lysine = 10 days. Batal & Parsons (2002).

A recent research was made by Batal & Parsons (2003) with broiler chick from 0 to 21 days, where the values of EM and the apparent digestibility of the amino acids from several protein sources were determined. The results showed a high digestibility of the

lacteous protein (casein) and a low digestibility of the soybean cake in the first week of life, possibly due to the presence of the oligosacharides, raffϊnose and stachiose. The processing of the soybean for protein concentrate and isolate improved the use of the nutrients by the chick, where the values of energy and digestibility of the amino acids from the protein isolate were higher than those from the concentrate, not only due to the reduction of the oligosacharides, but also due to the smallest potential for occurrence of a reaction of Mailard, which makes the complex lysine-carbohydrate unavailable for the metabolism of the bird, as demonstrated in table 10.

Table 10 - Effect of the age of the chick on the values of metabolizable energy (EM = Energia Metabolizάvet) and the coefficient of apparent digestibility of the lysine (%) in different sources of protein.

Source of Protein Age (days)

0-2 3-4 7 14 21

EM (kcal/kg)

Casein 3796 3751 3755 3857 3878

Soybean Powder 2783 2725 3034 3128 3089

Concentrate of Soybean Protein 3077 2852 3080 3278 3320

Isolate of Soybean Protein 3444 3280 3324 3609 3627

Coef. of Digestibility Applicable Lysine

Casein 97 97 97 98 98

Soybean Powder 77 79 89 87 84

Concentrate of Soybean Protein 83 16 91 94 94

Isolate of Soybean Protein 87 86 92 95 96

Batal & Parsons (2003)

Longo et al. (2003a), studying different protein ingredients in the pre-initial diets of broiler chicken, noticed that the animals that received as their main source of protein the protein isolate from soybean, or that had blood serum in their diet, had the lowest indexes of food conversion and exhibited, in absolute values, the highest gains of weight, as it is showed in table 11.

Table 11 - Performance of broiler chicken at 7 days of age fed with different sources of protein.

Parameters

Gain of Consumption of

Treatment CA. weight food

Corn + Soybean Powder 121.95 1.12 b 137 ab

Corn + Soybean Powder + cane yeast 119.73 1.21 c 144.2 a

Corn + Soybean Powder + Isolate of Soybean

123.77 1.01 a 124.2 c

Protein

Com + Soybean Powder + powdered egg 112.24 1.12 b 125.4 be

Corn + Soybean Powder + blood serum 122.47 1.05 a 128. .0 be

Corn + Soybean Powder + corn gluten 115.71 1.13 b 130. .2 be

Repeated averages of different letters in the same column differ among each other by the test of Tukey (p < 0.05)

Longo et al. (2003b) evaluating the EM of different sources of carbohydrates for broiler chick of 1 to 7 days, noticed that, except for lactose, the other ingredients had a high potential for supplying energy, as showed in table 12.

Table 12 - Values of EMAn (kcal/kg) for sources of carbohydrates determined in chick at the first.

EMAn

Corn Starch 3269

Manioc Starch 3690

Glucose 3427

Lactose 1225

Sucrose 3524

Batal & Parsons (2004) tested different sources of carbohydrates in diets with

Soybean Powder, and noticed that the use of sources of carbohydrates rapidly and highly digestible, consistently provided to the diets higher values of EMAn up to 7 days of age, as showed in table 13, as well as a larger gain of weight, as showed in table 14, with special attention to the ingredient Dextrose (monosaccharide).

Table 13 - Values of EMAn for chick of diets Soybean Powder-Source of Carbohydrate as a function of the age.

Source of carbohydrate Days of age

0 - 2 3 - 4 7 14 21

Dextrose 3,313 3,361 3,359 3,354 3,329

Corn Starch 3,208 3,079 3,109 3,238 3,254

Pre-gelatinized Manioc Starch 2,910 3,052 3,276 3,046 3,154

High Amylose Starch 1,992 2,132 2,384 2,096 2,292

Table 14 - Gain of weight of chick fed with different diets of Soybean Powder - Source of Carbohydrates.

Source of carbohydrate Gain of Weight (g)

Week l Week 0 - 3

Dextrose 88 a 461 a

Corn Starch 70 cd 380 bc

Pre-gelatinized Manioc Starch 69 cd 358 c

High Amylose Starch 60 cd 317 d

The broiler chick has difficulty in digesting saturated fat, due to an inefficient recycle of biliary salts. This lower digestibility of dietetic fat is caused mainly by the reduction in the digestibility of the saturated fat acids (16:0 and 18:0) (Penz Jr. & Vieira, 1998; Rostagno et al., 2000a).

Sakomura (1996) cited by Rostagno (2004) evaluated the effect of the age of the broiler chick, between the 1st and the 7th and the 22nd and 28th days, on the values of metabolizable energy (EMn) and the digestibility of fat from different types of soybean. The author noticed that the older birds exhibited higher levels of EMn and digestibility of fat. The lower value of EMn and digestibility of fat found in the soybean cake associated to oil, with respect to the extruded soybean, can be explained by the higher level of lecithin in the later, which favored the emulsification and the absorption of lipids, as showed in table 15.

Table 15 - Effect of the age of the bird on the values of EMAn and the digestibility of the fat in different types of soybean.

A se ( da vsl Digestibility of fat (%) EMAn (kcal/kg)

1 - 7 22 - 28 1 - 7 22 - 28

Soybean Powder + oil 80.7 93.5 3883 3956 Toasted Soybean 73.4 83.4 3724 3798

Extruded Soybean 85J 953 4079 4459

Sakomura (1996)

Nutritional Requirements:

A great evolution took place in the Brazilian tables for birds and pigs that were edited by Prof. Horacio Rostagno of UFV. In his first version (1983) there were no specific recommendations for the first phase of 1 to 7 days of age of broiler chicken, as it is the case in the NRC of birds of 1994. However, in the editions of 2000 and 2005 of the Brazilian tables, the requirements for 1 to 7 days were given. The recommended nutritional levels according to the Brazilian tables (Rostagno et al., 2000 and 2005) and by the NRC (1994) for broiler chicken in the pre-initial and initial phases are showed in table 16.

Table 16 - Nutritional levels recommended by different sources for the pre-initial and initial phases.

Nutrient Unit 1 - 7 days 8 - 21 days 1 to 21 days *

EM Kcal/kg 2960 3050 3200

Protein % 22.11 21.14 23

Calcium % 0.942 0.899 1

Available Phosphorous % 0.471 0.449 0.45

Total Lysine % 1.503 1.311 1.10

Digestible Lysine % 1.363 1.189

Met + Cis total % 1.067 0.931

Met + Cis digestible % 0.968 0.844 0.9

Rostagno et al. (2005). *NRC (1994)

Requirements of aminoacids:

The establishment of the nutritional requirements of aminoacids for broiler chicken is based on two important nutritional concepts, these being the expression of the requirements taking into consideration the digestibility of the amino acids, and the establishment of lysine as the standard aminoacid, so that the requirement of the other aminoacids is established from a relationship with the level of lysine - concept of ideal protein - as demonstrated in table 17.

However, three great particularities must be addressed as of the establishment of the requirement for amino acids for broiler chicken in the first week of their lives:

These relationships can be different when considered the period immediately after the eclosion, due to the differences in the growth rate and in the synthesis of tissues in the first days of life, as well as the interference with the nutrition that comes from the content of the vitelinic sac. Two other amino acids (Glycine and Proline) considered dismissible in the diet of adult birds are, however, essential for the chick, even though the glycine may be substituted by serine in the diet.

The digestibility of the amino acids in the food at the age of 1 to 7 days is not the same as that which is usually determined in birds with 14 to 25 days of age or in adult roosters, which are those informed in the tables of composition of the foods that are currently available.

Table 17 - Ideal Protein — Relationship digestion of amino acid / digestion of lysine for broiler chicken recommended for different ages.

Sources 1 2 3 4 5 6

Phase, days 1 - 21 1 - 21 1 - 14 0 - 21 AU 7 - 28

Lysine % 100 (1) 100 100 100 100 100

Met + Cys % 71 (70) (1) 72 74 70 73 70

Threonine % 59 (67) (1) 67 66 66 65 66

Tryptophan % 16 17 16 — 16 14

Arginine % 105 (105) (1) 105 105 125 105 108 1. Rostagno et. al. (2000); 2. Baker (1997); 3. Degussa (1996); 4. Lippens et al. (1997); 5. CVB (1996); Gruber (2002). The values inside parentheses are by Tejedor (2002). References 2 to 6 cited by Fisher (2002).

Butteri (2001), using male broiler chick, fed with diets based on corn, soybean powder and corn gluten (3.1%), with 22% of gross protein, 3050 kcal of EM and different levels of digestible lysine, found that the birds that are fed with the levels of 1.16% and

1.247% of lysine, exhibited the best results of weight gain for the period of 1 to 7 and 1 through 14 days of age, respectively, as showed in table 18.

Table 18 — Gain of weight and consumption of digestible lysine in broiler chick.

Lysine Dig. (%) 1 to 7 days 1 to 14 days

Weight Gain, g. Cons. Lysine, g. Weight Gain, g. Cons, lysine, g

1.073 97.78 b 1.049 312.43 b 5.027 1.160 103.30 a 1.233 329.39 a 5.707 1.247 106.40 a 1.289 317.26 b 5.872

Butteri (2001)

The quality of the environment where the birds are created also interfere in the productivity and in the nutritional requirements of the birds. This affirmative is confirmed by the work of Toledo (2003), which studied the effect of 5 levels of digestible lysine and of the environment, clean or dirty, without infection and with reused bed, on the performance of broiler chick, noticing that in the period from 1 to 11 days, the gain in weight in the clean environment was higher than that in the dirty environment. In this phase, the recommended level of digestible lysine was of 1.30 and 1.26% for the clean and dirty environments, respectively. In the period from 12 to 22 days, the clean environment also provided the highest gain, and the requirement of lysine was of 1.24% for the gain of weight and conversion of food. In the dirty environment the requirement was of 1.14% of digestible lysine for the same parameters.

Glycine and serine are considered essential amino acids for the performance of the chick, mainly for participating in the formation of body protein, uric acid, creatine and purines, therefore, it can be expected that its nutritional requirement also varies for the different ages. Ngo & Coon (1976), studying the supplementation of L-glycine for male broiler chick from 1 to 9 days, noticed that the birds that were fed with diets supplemented with 1.3% of glycine - 2.15% of total gly + ser - presented a larger gain of weight than the birds that were supplemented with lower levels, as showed in table 19. The authors suggest that the administration of an ideal level of glycine during the first days may reduce the requirement in the later phases. Coon et al. (1974) noticed serine can substitute glycine in the diets of the birds in the period of 1 to 10 days, as long as this substitution is made on an equimolar basis.

Table 19 - Gain of weight of chicks fed with different levels of supplementation of glycine in the period of 1 to 9 days of life.

Supplementation of glycine (%) Levels of gly -I- ser (%) Gain of weight g/bird/day

0.3 1.15 6.8 c 0.9 1.75 7.5 b 1.3 2.15 8.8 a

Ngo & Coon (1976)

According to Schutte et al. (1997), the diets for broiler chick based on corn and soybean powder cannot have less than 21% of protein, because the amino acids glycine + serine would then be limiting. These same authors estimated the requirement of glycine + serine in 1.85%, while the NRC (1994) recommends 1.25% and Rostagno (2000) 1.44% for the period of 1 to 21 days. However, Rostagno (2005), recommends 2.26% of glycine + serine for the phase of 1 to 7 days and 1.97% for 8 to 21 days.

Rostagno et al. (2002ab), studying the use of lower levels of gross protein - 18 and 19% - with respect to the control — 22% of gross protein - supplemented or not with synthetic amino acids, for broiler chicks from 8 to 21 days of age, found that inside the amino acids that were studied - arginine, glycine, glutamic acid, valine and isoleucine - independent of the level of protein that was used, there was a reduction in the performance of the birds in the absence of supplementation with glycine. The continuation of the study with broiler chicks of the same age, fed with diets containing different levels of glycine, showed that 2.11 of glycine + serine was the requirement for the phase from 8 to 21 days. In this way, the level of glycine + serine has to be taken into consideration as a possible limitation for the growth of birds fed with low protein in their diet (Rostagno et al., 2002c).

The ideal protein allows for an easy calculation of the requirements of all amino acids using as reference the level of lysine in the food. The levels of nutrients that resulted in optimal productivity of the broiler chicks - 1 - 11 days - according to Rostagno (2004) were: Gross protein, 21 - 25%; Digestible Lysine, 1.18 - 1.24%; Total Glycine + Serine, 2.10 - 2.30%; Sodium, 0.40%; nutrients expressed in percentage of digestible Lysine: Digestible Metionine + Cystine, 72%; Digestible Threonine, 67%; Digestible Tryptophan, 19% and Digestible Arginine, 108%.

Sklan & Noy (2003), however, defined that the requirement for total lysine of 1 to 7 days is of 1.08% and that of total sulfured amino acids is of 0.91%, which would result in a ratio of 84%.

Levels of gross protein and energy. Birds do not have a requirement of gross protein as such, but they need essential amino acids and a sufficient amount of non essential amino acids. The requirements of broiler chicken with respect to essential and non essential amino acids seem to increase as higher levels of gross protein are used in the food, suggesting a lower efficiency of the use of protein, associated to an unbalance of the amino acids. Metabolic studies indicate that an elevation in the level of protein in the ration stimulates protein catabolism, through a greater synthesis of pancreatic and intestinal enzymes and also of the enzymes that are involved in the degradation of the amino acids. In summary, the increase in the requirement of some essential amino acids can be explained, in part, by their lower conservation due to the use of high levels of protein in the diet. The excess of proteins - essential + non essential amino acids - is catabolized and excreted in the form of uric acid followed by an energetic cost for the bird (Sklan & Noy, 2004).

Experiments conducted at the Universidade Federal de Vicosa, in the decade of 1980, and by Costa (1981) indicate that it is possible to reduce the level of gross protein in the ration to 21.5% without affecting the gain of weight and the conversion of food of the broiler chicken.

Some authors argue that the use of a high level of gross protein is due to the fact that the birds in their first stage of life need an environment with a high temperature, thus, this excess of amino acids would serve to supply the birds in case of a probable lack of heat, as the protein catabolism results in the release of heat. Penz & Vieira (1998) report that the use of pre-initial diets with higher levels of protein has a positive effect in environments with temperatures that are below the comfort zone.

Rocha et al (1999), studying three levels of gross protein in the pre-initial diet concluded that higher levels of gross protein (26%), provide a better performance of the birds. Similarly, Toledo (2002) conducted two experiments splitting the period of 1 to 21 days into pre-initial phase - 1 to 7/10 days - and initial phase - 7/10 to 21 days - showing that the use of 30Og of a pre-initial diet with 25% of protein - 1.18% of digestible lysine - resulted in a

better gain of weight (754g) than the chicken that were fed with an initial diet of 22% of protein - 1.14% of digestible lysine 721 g.

Two experiments were conducted recently at the Universidade Federal de Vicosa by Toledo (2003), to investigate the effects of different levels of gross protein and the type of the environment, clean and dirty - without disinfection and with reused bed - using broiler chicks - 1 - 11 and 12 - 22 days - arriving to the conclusion that when adequate levels of amino acids are maintained, the gross protein in the pre-initial and initial rations can be reduced, respectively, to 21% and 20%. The birds that were created in the dirty environment exhibited a lower gain of weight during the 2 periods. Sklan & Noy (2003) also noticed positive effects on the gain of weight of chicks of high levels of protein in the pre-initial diet from 1 to 7 days of age. Rostagno (2005) recommends 2950 kcal of EMA for chicks with 1 to 7 days of age. Nascimento et al. 2005 noticed that the levels that are used by the industries varied between 2904 and 3040 kcal of EM, however, they did not check the effects of different levels of EM on the performance of chicks from 1 to 7 days of age. Levels of sodium.

Large variations have been seen in the literature regarding the nutritional recommendations for sodium. The NRC (1994) recommends the level of 0.20% for the period from 1 to 21 days. Murakami (2000) determined for this phase 0.250%, while Rostagno et al. (2000 and 2005) recommend 0.224% for the period of 1 to 7 days and 0.216% from 8 to 21 days. These values are much lower than the requirements determined for the phase from 1 to 7 days by authors such as Britton (1992), cited by Penz & Vieira (1998), which recommends 0.39% for the first week. Maiorka et al. (1998) suggest the level of 0.40% and highlight that this level does not present a negative effect on the quality of the bed; Vieira et al. (2000) recommend 0.36% for the first 7 days and Vieira et al (2003) recommend from 0.38 to 0.40% of sodium during the first week.

Chicks that were kept fasting exhibited low activity of Na+, K+ ATPase. The supply of sodium-rich diets stimulated the activity of this enzyme; however, diets that are poor in sodium depress the activity even more. The low absorptions of glucose and methionine that are noticed right after the eclosion are not only related to the hydrophobic nature of the vitelinic residue present in the intestine, but also to the low concentration of

sodium, as it has an important role in the absorption of these nutrients that have as their mechanism of absorption carriers that depend on sodium (Sklan & Noy, 2000).

The ingestion of water is directly dependent on the age of the bird and on the level of sodium in the diet. The increase of sodium in the diet during the first weeks of life of the birds increases the consumption of water and may compromise the quality of the bed, this effect being more pronounced during the third week. According to Penz & Vieira (1998), even though there is a production of excretion with greater level of humidity in the first seven days of life, the amount of excretion is small, and would have little effect on. the quality of the bed until day 21. Even though the relationship between the ions sodium, chlorine and potassium is important - the electrolytic balance, the experimental results are still contradictory, there being recommendations of 160 to 340 meq/kg of diet, the results, however, showing a trend towards a central value of 240 meq in the majority of the works (Martins, 2003; Vieira, 2004; Vieira 2005 and Borges, 2002). Grain size (granulometry) in the ration.

The rations used for broiler chicken are supplied either in the form of a powder, or in pellets, or extruded, or crushed. In case it is used in the powder form during the pre-initial phase, special attention must be given to the size of the grains of the ingredients, in order to allow for an adequate consumption and digestibility of the ingredients. On the other hand, there are clear evidences that the newly born broiler chicken have a preference towards crushed or pelletized rations.

According to Penz Jr. & Maiorka (1996), the nutritionists have a preference towards fine and uniformly ground ingredients, with the expectation that these are more easily digested by the enzymes that are present in the gastrointestinal tract. However, very fine particles usually adhere to the beak of the birds, reducing the consumption and increasing the losses, thus affecting the performance.

The determination of the grain size (granulometry) through its geometric diameter (DGM) seems to be the most efficient method in the characterization of the particles. Nir et al. (1994c) call the attention for the fact that the average geometric diameters of the rations may exhibit similar values; however, they may be composed by very different particles. Thus, in order to characterize the granulometry in an adequate way, it is necessary,

besides establishing the DGM, to establish the standard geometric deviation (DPG - Desvio Padrao Geomέtricό) of the particles, when a small value of DPG indicates greater uniformity. Nir et al. (1994c) performed a series of experiments seeking to identify the most adequate geometry for the birds in the pre-initial and initial phases. Table 20 contains a summary of the main recommendations of the authors for these phases. Similar values of grain size - 1050 to 1230 mm - for the pre-initial rations of broiler chicken were recommended by Toledo et al. (2001).

Table 20 - Average size of the particles in the diet or in the corn, recommended for broiler chicks in the pre-initial and initial phases.

Source Age (days) Grain Size (mm) Product

Nir et al. (1994a) I to 7 ns Diet

Nir et al. (1994a) l to 21 1.113 to 1.230 Diet

Nir et al. (1994c) l to 7 0.769 Corn

Nir et al. (1994c) 7 to 21 0.769 Corn

For trying to explain why the smallest particles do not improve the performance of the birds, research is being held with the purpose of relating the grain size to changes in the gastrointestinal tract of the birds.

Nir et al. (1994b) made an evaluation of the influence of the grain size on the gastrointestinal tract of broiler chicks in the period of 7 to 21 days, fed with rations having coarse, medium and fine grain sizes. Changes in the weight of the gizzard, of the duodenum, of the abdominal fat and of the pH of the digestive content of the gizzard and of the intestine were found, as showed in table 21. The changes that were noticed in the gastrointestinal tract suggest that grain size may interfere at the rate of passage. Large particles reduce the velocity of the passage, improving the digestion of the diet; whereas fine particles, due to their rapid transit, would lead to an atrophy of the gizzard, a slight hypertrophy and a low pH of the intestine, what at the end would affect the appetite of the birds and consequently their performance. The presence of very fine particles in the diets cause the formation of a paste and the accumulation in the beak, increasing the consumption of water and the loss of diet at the water fountain (Nir et al., 1994b).

Table 21. Effect of grain size on the gastrointestinal tract of broiler chicks in the period of 7 to 21 days of age.

Grain Size

Parameters

Fine Medium Coarse

Relative weight (g/100g of living weight)

Gizzard 2.22 c 2.80 b 3.13 a

Duodenum 1.25 a 0.89 b 1.07 b

Abdominal Fat 0.71 b 0.7S b 0.99 a pH of the gizzard 3.57 a 2.77 b 2.91 b pH of the intestine 5.97 b 6.23 a 6.35 a

Lόpes & BaiaO (2000) concluded that, for powdered rations, the coarsest grain size is the most indicated for the ingredients, as the increase in the diameter of the particle causes an increase in the weight of the gizzard. This effect is due to the higher activity of the muscles and the larger volume of diet present in this organ. Kilburn & Edwards (2004) checked that diets containing soybean particles of 1.239 mm of DGM provided higher gain of weight to the birds as compared to the diets that contained soybean particles with 0.891 mm of DGM.

Physical form of the ration.

Nir et al. (1990) observed that when the birds can choose freely between the several physical forms of the diet, they prefer the pelletized diet and exhibit a better performance, a fact which can be explained by a reduction in the energy that is needed for their maintenance, due to the smallest amount of movement that they have to make during the consumption of the pelletized diet. Agreeing with this affirmative, Leeson & Summers (1997) reported that for the consumption of one same amount of diet, the birds that consumed it in the form of a powder spent a time that was three times larger as compared to the birds that received the diet in the form of pellets. Reece et al. (1986), working with broiler chicks from 1 to 21 days of age, used two physical forms of the diet - powdered and crushed, and concluded that the birds that received the crushed diet had a larger gain of weight as compared to those that received the powdered diet. The authors noticed that this difference in the gain of weight was maintained until the 47th day. Zanoto et al. (1996) evaluated two physical forms of diet - powdered and

crushed - for broiler chicks from 1 to 21 days. The crushed diet provided a higher body weight, a larger consumption of the diet and a better conversion at the 21 days of age, this difference in the gain persisted until the end of the experimental period.

Souza et al. (2004) evaluated different physical forms of pre-initial diets and noticed that the birds has a worse performance when they received the diet in the for of a powder, the best results being reached with the use of a diet in the crushed form, as showed in table 22.

Table 22 - Performance of broiler chicken at the 7 days of age fed according to the physical form of the ration.

Parameters Parameters

Gain of Weight CA. Consumption of Ration

Physical form of the ration

Crushed 156.73 a 1.302 c 204 b

Micro-pelletized 149.77 a 1.374 b 206 b

Powdered 125.18 c 1.766 a 221 a

_____

The quality of the ready ration is also an important factor. The monitoring of the resistance of the pellet and of the presence of fines is a capital issue for the gains obtained in the food conversion not to be lost, mainly if the ration is delivered in the bulk state by trucks.

Nutrition technologies for the first week. Non conventional

An alternative to the use of pre-initial diets is the use of nutritive solutions that can be fed to the birds via intubation at the incubatory or through the drinking water. Working with the second option, Toledo (2002) concluded that the use o nutritive solutions administered to the birds to the seventh day in the drinking water, associated to the powdered diet, improved significantly the weight of the birds at the 21 days, and this difference was maintained through the 40 days of age. In another experiment, the same author tested the association of the nutritive solution with the use of the pelletized diet, and noticed that the

solution was effective in the improvement of the gain of weight of the birds in the period of 1 to 21 days, as showed in 23.

Table 23 - Effect of the supply of the nutritive solution via drinking water - 7 days - on the performance of the broiler chicken receiving powdered and pelletized diets.

Experiment ^' 1 - Powdered

Experiment 2 - Pelletized Ration Ration

Nutritive Solution

Food

Gain of weight (g) Gain of weight (g) Conversion g/g

1 - 21 days 1 - 40 days 1 - 12 days 1 - 12 days

Without 640 2139 275 1.102 With 671 2174 290 0.975

Another option that is being studied to guarantee the adequate nutrition of the newly born birds is pre-lodging feeding, that is, the supply of the food to the chicks while they are still in the incubator, with the purpose of maximizing the gain of weight, once from the incubator to the chicken farm, in the average, the chicks spend about 10 to 36 hours, a time that is needed for the practices of vaccination, separation according to the sex, and the transportation itself. Noy & Pinchasov (1993) made an experiment where one group of chicks received, at the incubator, via intubation, a nutritive solution. The birds that had been intubated with the nutritive solution and that had received immediate access to the diet exhibited better performance, as showed in table 24.

Table 24 - Effect of the supply of nutritive solution via intubation, made at the incubator, on the performance of the broiler chicken in the period from 1 to 40 days.

Group Nutritive Solution Fasting (24 hours) Gain of Weight (g)

1 Yes No 2032

2 Yes Yes 1912

3 No No 1915

4 No Yes 1805

Noy & Sklan (1999) noticed that the supply of solid, semi-solid or liquid diets, immediately after the eclosion, provided a larger gain of weight than the animals that were maintained fasting until their lodging, after 34 hours. The oral administration of 0.4 mL of nutritive solution was capable of allowing a gain of weight similar to the treatment with food at will. They also noticed that feeding right after the eclosion, still in the incubator, provided an increase in the percentage of breast in the carcass.

Toledo (2003) evaluated the effects of the feeding of female broiler chicks with a humid diet - crushed or pelletized diet + 20% of water - placed inside transportation boxes (2g/bird). The birds remained for approximately 20 hours inside the transportation boxes and were transferred to the pre-experimental shed where it was determined the performance to the 10th day of life. The chicks that were fed with the humid diet in the boxes showed better productivity than the control group - which was fasting, as showed on table 25.

Table 25 - Effect of the supply of a humid diet to broiler chicks during the permanence in the transportation boxes on the performance at the 10th day.

Diets in box (20 hours) Weight gain (g) Dietary conversion (g/g)

Control (fasting) ϊ7ϊ 1,419

Wet crushed 187 1,330

Micropelletized (wet) 180 1,351

Toledo (2003).

At last, feeding in egg is no longer a very distant reality, only missing a greater volume of research, in the area to consolidate already promising results. Tako et al. (2004) and Uni et al (2005) showed that it was possible with administration in egg of carbohydrates and Beta-hydroxy-beta-methylbutyrate to stimulate the embryo's intestinal development so being to improve the chick's weight gain and increase the chest muscle percentage in the carcass. Recommendations of special ingredients in pre-initial diets. Recommendations about using food in pre-initial diets of meat producing chickens are: reduction of corn and soy bran and inclusion of high-nutritional value ingredients - high digestibility - such as glucose, blood plasma, fish flour, corn gluten bran and soybean protean concentrate or soybean isolated. Wheat, rye and barley must be avoided

because they increase intestinal viscosity and reduce digestibility or nutrients. High levels of oil are not recommended, but incorporation of 1 to 2% of oil may be beneficial because it reduces pulverulence of the food.

The use of blood plasma has been studied as a mechanism to help intestinal immunity by the presence of antibodies in its composition, in addition to having a high protein contents, rich in essential high-digestibility amino acids, essential to the bird's muscular development. . Results of experiments conducted with blood plasma in bird nourishment show beneficial effects of using this ingredient on bird performance (Rostagno et al. 2004). However, statistical differences have shown only in higher ages - 21 and 42 days - and with continued use of the ingredient.

Sources o easily digested carbohydrates are also important. However, corn in grain is already rich in high-digestibility carbohydrates. As commented before, chicks after hatching may not have capability to digest with efficiency large amounts of maltose and disaccharides - the main carbohydrates formed in starch digestion process, and therefore supply of promptly available monosaccharide for absorption is an option that has demonstrated good results, they are high-cost ingredients (Batal & Parsons, 2004).

Phospholipids and cholesterol esters represent about 1 third of the yolk sac lipids, the remainder being represented by triglycerides. The synthesis in the cholesterol organism and phospholipids requires considerable energy spending, and, both are important components in the composition of cell membranes in addition to acting in lipid transport processes in the circulation. Under the biochemical point of view, it would be extremely ineffective to catabolize phospholipids and cholesterol, in order to re-synthesize them later.

The energy released by consuming all triglycerides released by the yolk sac on the first day - considering a 100% efficiency in theirs utilization - would be approximately 9 kcal, lower therefore than the estimated 11 kcal as a maintenance en energy requirement for the first day of the bird's life only, therefore, even triglyceride reserves must be seen as help in maintaining the chick' s life and not as the main food in the post-hatching period.

In the newly-hatched chick's liver, over 80% of lipids are found under the form of cholesterol esters. Cholesterol esterification may be interpreted in a relatively non-toxic manner to store great amounts of cholesterol for using in lipid transport in embryogenesis.

After hatching, cholesterol may be used as a membrane structural component or further functionally in lipid transport in the circulation.

The remaining 20% of lipids found in the liver 4% are triglycerides and 14% phospholipids, both rich in arachidonic and docosahexaenoic acid (DHA), which have their synthesis in the yolk sac membrane cells from other fatty acids. Arachidonic and DHA acids may modify the metabolism of eicosanoid acids, thus affecting immune and anti- inflammatory responses in the neonate. Further, DHA acid is the most used fatty acid in forming the central nervous system and the embryo's retina, having, therefore, greater value for this purpose than supplying energy to the embryo.

At last we can assume that during the evolutionary process, newly-hatched chicks normally having easy access to exogenous foods, it has provided the check with specific macromolecule reserves that would hardly be found in nature, as is the case of specific fatty acids - arachidonic and DHA, cholesterol and phospholipids present in the egg.

The egg incubation process lasts approximately 21 days. This time is, however, influenced by factors such as egg storage period and room temperature in the pre-incubation period, incubation conditions — especially temperature and relative humidity in incubators, genetic lineages and matrix age, etc.

Due to these factors, not seldom variations greater than 24 hours between the chick births of the same flock are observed in incubators, and this variation can easily reach

48 hours. This means greater staying time in the birthplace of eggs that are hatched earlier.

Invariably, these chicks will suffer, in lesser or greater degree, a dehydration process that starts 2 hours after eggs are hatched.

Such fact can be further aggravated if the incubation process, low relative humidity periods occurred for long periods, or if chicks come from young matrixes, thus having less weight when they are born.

From leaving the birthplace to the placement, the chicks are further classified, vaccinated, sexed and housed in transport crates. It is desirable that after housed in the crates, the birds be sent to the poultry farm on the same day, however, not seldom are cases verified in which the birds stay overnight in incubators to be delivered on the following day.

The distance from the incubator to poultry houses, the quality of roads, and, the quality of the delivery truck may further determine a higher time than 10 hours during transportation.

All these delays may imply that the birds may be housed about 48 to 72 hours after their birth. During this period the chicks lose weight by using nutrients from the yolk sac, by digestive and renal excretions, and dehydration.

Lodging the chicks with access to water and food must be as quick as possible, respecting the minimum necessary time in the birthplace. Weight loss and dehydration caused by fasting, even if minimum, may cause an increase in the death rate, intestinal mucous development delay, causing lesser efficiency in digesting and absorbing nutrients. Old handling recommendations mentioned fasting as a practice for performance improvement, because it would favor a faster reabsorption of the residual yolk sac. Such practice has proved to be inadequate since it has been widely demonstrated that it is exogenous food ingestion that accelerates utilization of the residual yolk sac.

Noy & Sklan (2000) verified that chicks deprived of exogenous food for 48 hours after hatching suffer weight reduction. However, during these 48 hours the small intestine weight increases 60% in chicks devoid of food and 200% in chicks provided with food. Controlled experiments have shown that supplying a nourishing solution in watering places ahs increase the performance of meat producing chickens in comparison that birds that have not received this supplement. The use of liquid-nourishment supplements in this final phase of incubation is, however considered as a risk to biosafety, by many incubators, in addition to establishing a need for adaptation of current equipment - birthplaces.

In the period immediately after hatching, the chick's intestine weight increases more quickly than its body weight as a whole. This rapid development process reaches a peak around 6 to 8 days for the intestine; however, other digestive tract organs such as the pancreas and the gizzard do not present the same pace of growth as to their relative weight. Noy & Sklan (1997) verified that weight increase in chicks only occurs 36 to 48 hours after having access to the diet. Growth start may be advanced by the precocity of access to the diet.

The benefit of this consumption advance is shown to be more pronounced in weight after 7 and 10 days of age, being that obtained advantage is maintained up to slaughtering. Required maintenance energy for the chick in the first 24 hours has been estimated at approximately 11 kcal (112 kcal. W 0.75). Assuming that the entire yolk sac residual contents released in the first 24 hours will be used as a source of energy with 100%

of efficiency, we would only have 9.4 kcal. Thus, without an additional nutrient supply will go into a negative energy balance and will certainly lose weight (Dibner et al. 2005)

In addition to weight loss observed in chicks submitted to fasting over 72 hours after birth, a high death rate can be observed in the flock in the first week of lodging, and low vitality of surviving chicks.

Development of digestive tract organs of chicks after hatching has suffered a strong influence of selection processes. High priority must be given to digestive, circulatory and respiratory systems, however it is supposed that nutrient partition for development of organs and tissues related to the immune response has been impaired. Depriving the chick of food right after hatching brings on reduction in the

Bursa weight, sharper than body mass loss itself. Food supply on the 3 ^rd post-hatching day does not correct this loss, the Bursa persisting with smaller size at least up to 21 days of age.

Dibner et al. (1998) demonstrated that chicks fed with a hydrated nutritional supplement presented high proliferation of lymphocytes in the Bursa 3 days after hatching. To the contrary, chicks kept in fasting presented absence of lymphocytes, demonstrating that the yolk sac residual contents present in the chick after hatching does not serve as a substitute to exogenous nourishment.

In order to soften the problem with lodging birds that already arrive weakened at poultry poultry houses, the barns should be suitably prepared at least 24 hours in advance and specific measures must be adopted to improve performance of flocks.

- The barn must be previously heated, cleaned and disinfected.

- The bed to receive weakened flocks of chicks should be new

- The barn should have good sealing air currents from getting in

- Supply drinking fountains and feeding places in sufficient amount - Water temperature may not be less than 18 ⁰ C, and if possible, a soluble poly vitamin should be added (e.g., soluble)

- Offered fed should preferable be special, appropriate for the post-lodging immediate phase. The use of offered feet on sheets of paper, helps the birds to begin their feeding more quickly, also preventing the chicks from ingesting bed particles; - The chicks should be lodged with maximum care, mainly in their handling, but quickly.

- Keep full attention, monitoring the internal barn environment and behavior of birds, especially in the first week of life, immediately reporting abnormalities to the technical responsible that serves you.

- Provide the barn with adequate lighting and ventilation - air renewal. The best strategy to maximize chick growth is certainly to provide them with food as quickly as possible. Nutrient ingestion stimulates the development of digestive organs, the immune system and utilization of yolk residue. Feeding in the first hour of life has unquestionable advantages that persist chicks are slaughtered.

In general, complete adaptation of the chick's digestive tract and metabolism to using a diet rich in carbohydrates and relatively poor in fat in replacement to using the yolk sac residue, lasts between 3 to 4 days.

The chick's digestive tract in the immediate pre-hatching moment has only the yolk sac residue available to perform digestion and absorption processes. This substrate is rich in fats and proteins and almost absent in carbohydrates, therefore with a high hydrophobic characteristic. Feeding at the post-hatch moment for being rich in carbohydrates, in addition to having little hydrophobic characteristic, needs digestive enzymes before unnecessary as amylase, maltase and others. In this phase, it is indispensable to use a special diet in order to maximize bird performance.

This special diet containing high-digestibility ingredients, aimed at facilitating transition from embryonic metabolism to post-hatching metabolism, by using adequate nutritional levels, sizes of particles and appropriate physical shape are not luxury spending, but rather a powerful technology that is found on the market today.

Currently, the herein applicant is the only company in the market to make a ready, crushed feed, specifically developed for the first four days of life, which will stimulate bird growth, development of the immune system and chicks' vitality increase, and also to offer an appropriate product to be supplied in the chick's transportation box to poultry poultry houses in the proportion of 5 grams per bird.

Therefore, the best strategy to maximize chick growth is to provide them with food as soon as possible. Ingestion of nutrients stimulate the development of immune system digestive organs and using the yolk sac residue. Feeding in the first hours of life has unquestionable advantages that persist until slaughter.

Egg nutrition still has ahead of it a huge of research so that its potential may be used the maximum.

The study of ingredient nutritional value for chicks is another field that must be quite developed, in order to be able to expand the current data base so that we will have more safety in formulating diets for this phase.

The basic invention form consists of a palletized feed and crushed with a particle size close to 2 mm, where feeds are removed using sieves. In another modality of the invention, the feed may be presented in micropelletized, microextruded or pressed forms .

The following researches have been used in developing the product or testing finished products :

Nutritional composition of foods for chicks from 1 to 4 days.

The product nutritional composition was developed through a series of metabolism tests aimed at studying the values of metabolizable energy and digestible nutrients of ingredients for chicks from 1 to 4 days old. Nutritional needs of chicks from 1 to 4 days old were determined based on nutritional values of ingredients determined in 1 to 4 day-old chicks - previous experiment - using for such experimental models of chick performance response to growing doses of nutrients in the diet.

Determination of the nutritional value of foods for 1 to 4-day old chicks The product nutritional composition was developed through a series of metabolism tests aimed at studying metabolizable energy values and digestible nutrients of ingredients for 1 to 4-day old chicks. Using controlled food methodology, and total excrete collection, several ingredients had their nutritional values determined. For each ingredient 6 cages containing 12 chicks each were used, which received feed consisting of a reference diet containing from 10 to 40% of the test ingredient. Concurrently, the test feed itself and a group of fasting birds were submitted to total excrete collection in order to determine the nutritional value of the test diet and endogenous losses, in order to allow determination of test ingredient nutritional value.

EMVn and digestible amino acid values. Values for chicks from 1 to 4 day old are found in the tables below:

Experiment 2

Fasting period evaluation and of different types of feeds, under performance and immunological characteristics of meat producing chickens.

5 The purpose of this experiment was to evaluate the immunological aspect of birds that consumed the nutritional formulation of this invention. Material and Methods:

150 Cobb 500 lineage meat producing chickens originating from 37-week old matrixes were used. These birds were lodged with an average weight of 37.5 grams. For this 0 experiment the birds were only vaccinated against Marek disease in the incubator.

These birds were separated into three treatments, being that in the first one the birds consumed 10 days of a standard bran pre-initial diet; in the second the birds received the nutritional formulation of this invention - 5g consumption - being supplemented with a bran pre-initial diet up to 10 days; in the third treatment the birds remained in alimentary fast for 5 36 hours prior to lodging and right after the lodging a standard bran pre-initial diet was supplied.

After the lodging four birds of each treatment were daily slaughtered up to the twentieth day, aimed at collecting cecal tonsils for checking germinative centers. On the fifth and fifteenth days of age 20 birds of each treatment were slaughtered to collect Fabricious 0 Bursa aimed at evaluating the percentage of lymphocytes per diagnosis per image.

On the twentieth day the remaining birds were vaccinated for New Castle disease via ocular, being submitted to blood collection at 40 days of age. This material was submitted to a histopathology laboratory for evaluation of vaccinal titles by serology through Elisa test.

Results were analyzed by SAEG (2000) program following an entirely randomized delineation, averages being compared by Tukey test at 5% of likelihood. Obtained Results:

TABLE 1. Weight gain of one to 21 days old meat producing chickens.

Treatment GP 1 A 7 GP 7 A 21

10 days of Bran Feed 175.4b 629.5b

Nutritional Formulation 180.1a 647.5a

36 hrs of Fasting + Bran 160.4c 544.4c TABLE 2. Bursa weight and lymphocyte percentage of 5-day old meat producing chickens.

Treatment Bursa weight, g % Lymphocytes

10 days of Bran Feed 0.1562 ^a 31.92b

Nutritional Formulation 0.1813 ^a 33.67 ^a

36 hrs of Fasting + Bran 0.1025b 29.69c

TABLE 3. Bursa weight and lymphocyte percentage of 15-day old meat producing chickens

Treatment Bursa weight, g % Lymphocytes

10 days of Bran Feed 0.95S8 ^a 42.50 ^a

Nutritional Formulation 1.1444 ^a 41.41 ^s

36 hrs of Fasting + Bran 0.7262b 39.10 ^a

TABLE 4. Elisa Test for New Castle Disease obtained with 40-day old meat producing chickens.

^" Treatment AMT GMT CV7%

10 days of Bran Feed 2875 1967 90.9

Nutritional Formulation 3226 2156 82.5

36 hrs of Fasting + Bran 2610 1896 115.7

Where: AMT = Mean Arithmetic Title and GMT = Mean Geometrical Title

Conclusions.

- The alimentary fast compromised the birds' performance and immunologic aspects. The birds that received the nutritional formulation presented greater weight gain compared to birds fed with the bran diet only - At 5 days of age, the birds that received the nutritional formulation of this invention presented a higher percentage of lymphocytes in bursal follicles, mainly at 5 days of age;

- The birds that received the nutritional formulation of this invention presented a higher amount of germinative centers and at more precocious ages, demonstrating advances maturation of the immune system; - HI titles for New Castle were higher and the antibody variation coefficient was lower in birds that received the nutritional formulation of this invention, demonstrating a better uniformity in vaccinal response and better response of the immune system to induced challenges. Results of the new experiment Nutritional formulation evaluation of the invention under the performance and morphometry of meat producing chicken organ.

The purpose of this experiment was to evaluate, in two tests, the performance and morphometric parameters, in post-hatching phase, packed in transport cases with different diets in pre-initial phase. Two experimental tests were conducted using for it 416 meat producing chicks from a commercial lineage - Cobb 500. in both tests the birds were distributed in a factorial delineation 2 - birds fed or not with 5 grams of the nutritional formulation of this invention followed by a bran feed, following normal consumption recommendations of each diet.

These birds were directly purchased in a poultry farm, transportation begun right after vaccination against Marek, and where the animals were provided with the nutritional formulation. From bird removal period to lodging in cages 24 hours were respected, average period of the incubator-barn period in normal conditions - counting the wait in the birthplace, processing period in incubator, bird rest, shipping and transportation.

After the 7 ^th day of age all birds consumed an initial and conventional growth diet.

In the first test the performance - weight gain, feed consumption and food conversion rate were evaluated - of 192 birds distributed, following an entirely randomized

delineation, in 32 cages - 8 repetitions / treatment - and the birds and feed being weighed in the lodging, 7, 14, 21, 28 and 35 days of age.

While in the second test morphometry - live bird weight, heart, liver small and large intestine, pancreas and bursa were evaluated - of 224 birds distributed, following an entirely randomized delineation, in 32 cages - 8 repetitions / treatment. One bird of each repetition was chosen and sacrificed on the 7 ^th , 14 ^th , 21 ^st , 28 ^th and 35 days of age.

Results we ^' re analyzed by SAEG (2000) program following a factorial analysis of 2 birds fed or not with the nutritional formulation of this invention in transportation case phase, and 2 birds fed with the nutritional formulation of this invention and bran feed, following normal consumption recommendations of each diet, averages being compared by Tukey test at 5% likelihood. Live Weight TREAT LIVE WEIGHT - G

DAYl DAY 7 DAY 14* DAY 22 DAY 28 DAY 35 CASE

With Nutritional

Formulation 40.01a 170.20a 468.84a 997.89a 1490.44 2112.96a

Without

Nutritional

Formulation 39.36b 162.60b 458.05b 974.33b 1464.07 2064.09b

CV, % 1^263 3,952 4,710 5,465 4,354 4,264

^" *PV 22 days - CASE P=0.063

Weight Gain

TREAT WEIGHT GAIN - G

14 TO 7 TO 22 TO

1 TO 7 7 TO 14 22 22 28 28 TO 35 22 TO 35 1 TO 35

CASE

With Nutritional

Formulation 130.21a 298.64 529.05 827.69 492.55 622.52 1115.06 2072.96a

Without

Nutritional 123.24b 295.46 516.28 811.74 489.74 600.02 1089.75 2024.73b

Formulation

CV, % 5,126 5,946 8,616 5,409 10,739 11,329 8,163 4,340

Feed Consumption

TREAT FEED CONSUMPTION - G

I TO 7 7 TO 14 22 7 TO 22 28 28 TO 35 22 TO 35 1 TO 35

CASE

With Nutritional

Formulation 133.41a 367.68a 794.10 1161.78a 815.02 1072.87 1887.88 3183.08

Without

Nutritional

Formulation 12,41b 356.50b 764.83 1121.32b 817.21 1093.28 1910.48 3158.22

CV, % 5 _; ,881 5,094 7,787 6,235 9,330 1 1,820 6,452 3 ,786

Food Conversion

TREAT FOOD CONVERSION - G/G

1 TO 7 7 TO 14 14 TO 22 7 TO 22 22 TO 28 28 TO 35 22 TO 35 1 TO 35

CASE

With Nutritional

Formulation 1.029 1.233 1.503 1.404a 1.657 1.731b 1.697b 1.537b

Without

Nutritional

Formulation 1 .027 1.207 1.483 1.382b 1 .676 1.838a 1.760a 1 .562a

CV, % 4 ,549 3,432 3,952 2,858 7 ,187 11,47 7,936 4 ,371

*CA 1 TO 35 days - CASE P=0.064

Morphometry: Day 3

RELATIVE MORPHOMETRY TO LIVE WEIGHT - DAY 03. %

RR RR

PV, PR PR FIG ₅ PR ID, PR IG, PR S. PR

TREAT MOEL, PANC,

G COR, % % % % Vit, % CAR, %

% %

CASE

With

Nutritional

Formulation 83.75 0.70 10.06 0.45 3.95 10.02 1.91 0.89 72.03

Without

Nutritional

Formulation 80.60 0.68 10.63 0.42 3.86 9.83 2.09 0.83 71.66

CV, % 12.136 10.342 13.282 19.138 21.494 18.582 22.118 49.650 5.080

Cor=heart; moel = gizzard; panc=pancreas; fig - liver; id=Small Intestine; ig=Large intestine; s.vit=yolk sac; car=carcass

Day 7

RELATIVE MORPHOMETRY TO LIVE WEIGHT - DAY 07. % __ _ _

PV, PR PR FIG ₅ PR ID, PR IG ₅ PR

TREAT MOEL, PANC, Bursa,

G COR, % % % % CAR ₅ %

% % %

CASE

With

Nutritional

Formulation 173.55 0.82a 8.04 0.59 4.39 11.72 1.64 0.15a 72.65

Without

Nutritional

Formulation 165.42 0.79b 8.01 0.61 4.24 12.08 1.61 0.13b 72.55

CV, % 9.832 9.334 10.992 18.199 9.809 9.060 17.031 22.570 2.186

Cor=heart; moel = gizzard; panc=pancreas; fig - liver; id=Small Intestine; ig=Large intestine; s.vit=yolk sac; car=carcass Day 14

RELATIVE MORPHOMETRY TO LIVE WEIGHT - DAY 14, % __ _ . _

PV, PR PR FIG ₅ PR ID, PR IG, PR

TREAT MOEL, PANC, Bursa,

G COR, % % % % CAR, %

% % %

CASE

With

Nutritional

Formulation 466.95 0.82 5.51 0.43 3.80 8.34 1.27b 0.20 79.62

Without

Nutritional

Formulation 45.43 0.79 5.72 0.43 3.73 8.40 1.36a 0.20 79.36

CV, % 11.225 10.928 13.506 18.905 11.392 14.445 25.146 21.496 2.762

Cor=heart; moel = gizzard; panc=pancreas; fig = liver; id=Small Intestine; ig=Large intestine; s.vit=yolk sac; car=carcass

Day 22

RELATIVE MORPHOMETRY TO LIVE WEIGHT - DAY 22, %

TREAT

CASE

With

Nutritional

Formulation 947.17 0.70 4.33 0.34 3.13 6.16a 0.98a 0.28 84.07 Without Nutritional

Formulation 924.97 0.69 4.27 0.31 3.05 5.72b 0.90b 0.28 84.79 CV, % 8.082 16.198 10.290 11.107 9.263 10.018 21.643 28.566 1.321

Cor=heart; moel = gizzard; panc=pancreas; fig = liver; id=Small Intestine; ig=Large intestine; s.vit=yolk sac; car=carcass Day 35

RELATIVE MORPHOMETRY TO LIVE WEIGHT - DAY 35, %

TREAT

_____

With 2182.08 0.50 3.19 0.22 2.19 4.40 0.95 0.24 88.31

Nutritional

Formulation

Without

Nutritional

Formulation 2084.38 0.50 3.37 0.22 2.20 4.19 0.89 0.22 88.40

CV, % 7.520 9.691 15.332 14.501 8.884 14.398 17.701 31.869 1.214

Cor=heart; moel = gizzard; panc=pancreas; fig = liver; id=Small Intestine; ig=Large intestine; s.vit=yolk sac; car=carcass ASCITIC RATE:

ASCITIC RATE AT 35 DAYS OF AGE

TREAT RELATIVE WEIGHT TO RIGHT RELATIVE WEIGHT TO LEFT VENTRICLE - ASCITIC RATE VENTRICLE

CASE

Conclusions

Using the nutritional formulation provided greater weight gain and better food conversion of birds at the end of the first week of life, such result and persisted up to 35 days of age.

The heart and Bursa relative weights were greater at the end of the first week in birds that received the nutritional formulation in the transport case. At 35 days of age the birds fed with the nutritional formulation presented a lower relative weight of the right ventricle, thus reducing the risk of ascites in birds, remembering that according to Aureliano 200O ₅ the birds can be considered ascetic when the right ventricle relative weight is higher by 40% in relation to the heart weight.

BIBLIOGRAPHIC REFERENCES:

Araύjo, L.F.; Nutricao Pόs-Eclosao: Aspectos Teόricos e Praticos. In: Anais do II Simpόsio sobre nutricao de aves e suinos. CBNA. Cascavel - PR. 2004. Batal, A.B.; Parsons, C. M. Effect of age on nutrient digestibility in chicks fed different types.

Poultry Science 81:400-407. 2002.

Batal, A. B.; Parsons, C. M. Utilization of different soy products as affected by age in chicks.

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