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Use of Betaine in Post-Hatch Feed for Broiler Chicks

ABSTRACT

Post-hatch delayed placement damages the physical and physiological development of broiler chicks. This study was designed to find adequate levels of betaine inclusion in pre-hatching and pre-starter feed, in order to minimize the negative effects of post-hatch delayed placement on broiler chicks. Newly-hatched chicks were allotted in a completely randomized design, with five treatments, five replicates of ten birds each. Five dietetic levels of betaine (control, 0.070, 0.130, 0.200 and 0.260%) were used in the pre-starter ration, offered to the chicks in the transporting box and during the pre-starter phase. Performance, yolk sac retraction, plasma glucose concentration, weight and histomorphometry of the small intestine were evaluated, after 24 hours of feed access and at 7 days of age. A metabolic trial was performed from seven to ten days of age. Betaine supplementation linearly influenced the chick’s ileum crypt depth after 24 hours in the transportation box. There was a quadratic effect, with an increase in feed intake up to 0.152% betaine supplementation. There was an improvement in the quality of the jejunum with betaine supplementation above 0.1%. The metabolizable coefficient of the ether extract was improved until reaching 0.163% of betaine supplementation. Betaine supplementation around 0.150% increases feed intake, the use of ether extract and interferes with the intestinal villi of chicks at seven days of age.

Keywords:
Delayed placement; glucose; histomorphometry; poultry

INTRODUCTION

The post-hatch period is considered a critical time for broiler chicks. It is a phase of adaptation to the external environment to the egg and specially to feed coming from exogenous sources. What makes this period even more difficult is the fasting that these chicks undergo.

Because of standard incubator procedures, chicks may not have access to feed for approximately 48 hours, from their hatching until their placement, which could mean they do not have access to feed for two or three days (Bhuiyan et al., 2011Bhuiyan MM, Gao F, Chee SH, Iji PA. Minimising weight loss in new broiler hatchlings through early feeding of simple sugars. Animal Poultry Science 2011;51(11):1002-1007.). Delayed intake of feed can hinder physiological and immunological development, reducing the overall productivity of chick-rearing (Van Den Brand et al., 2010).

Specific formulas were recommended for the post-hatch and pre-starter phases diets of broiler chicks. These formulas include the presence of trophic agents that can attenuate the effects of post-hatch delayed placement, to increase precocity in the physical and physiological development of broiler chicks.

Among these trophic agents is betaine. According to Ahmed et al. (2018Ahmed MMM, Ismail ZSH, Abdel-Wareth AAA. Application of betaine as feed additives in poultry nutrition - a review. Journal of Experimental and Applied Animal Sciences 2018;2:266-272.), betaine is not present in large quantities in poultry feedstuffs such as corn and soybean. Dietary betaine supplementation has many benefits for the productive performance and health status of poultry. The primary function of betaine is as a donor source of methyl groups, for example, methylation of DNA, RNA and lipid cell membranes to the synthesis of methionine, carnitine and creatine. In broiler diets with methionine deficiency, betaine supplementation improved the metabolizability of ether extract and protein (El-Husseiny et al., 2007El-Husseiny OM, Abo-El-Ella MA, Abd-Elsamee MO, Ab-Elfattah MM. Response of broilers performance to dietary betaine and folic acid at different methionine levels. International Journal of Poultry Science 2007;6:515-523.).

Also, betaine is an organic osmolyte. Unlike inorganic salts, osmoregulatory substances are soluble and remain in high concentrations inside cells without impairing cell metabolism (Kempf & Bremer, 1998Kempf B, Bremer E. Uptake and synthesis of compatible solutes as microbial stress responses to highosmolality environments. Archives of Microbiology 1998;170:319-330.). This mechanism is very important for intestinal cells to maintain the exchange of water and solutes during digestion and absorption of nutrients. There are several molecules considered organic osmolytes, such as proline, taurine, glutamine and glycine, but betaine is considered the most effective (Jahn et al., 2006Jahn MP, Cavagni GM, Souza DK, Kucharski LC. Osmotic effect of choline and glycine betaine on the gills and hepatopancreas of the Chasmagnathus granulata crab submitted to hyperosmotic stress. Journal of Experimental Marine Biology and Ecology 2006;334:1-9.). Betaine also has the function of preventing dehydration of bacteria, such as enterobacteria and lactobacilli, in hyperosmotic environments (Li et al., 2015Li C, Sun J, Qi X, Liu L. NaCl stress impact on the key enzymes in glycolysis from Lactobacillus bulgaricus during freeze-drying. Brazilian Journal of Microbiology 2015;46(4):1193-1199.). Therefore, betaine supplementation in the broiler diet appears to be an alternative to improve mucosa development in the first post-hatch week, which is also a challenging time for the chick.

The objective of the present study was to ascertain the appropriate levels of betaine inclusion in post-hatch and pre-starter feed, to minimize the negative effects of post-hatch delayed placement and ensuring intestinal integrity and nutritional support for these chicks.

MATERIALS AND METHODS

This research on animals was conducted according to the institutional committee on animal use under protocol nº 008/13. A total of 1,500 Cobb 500 male chicks originated from eggs produced by breeder hens the same age and incubated in the same machine were used.

The treatments consisted of five supplementation levels of Betaine HCl 95% (72% de betaine): Control (no supplementation); 0.070, 0.130, 0.200 and 0.260% inclusion per tonne of pre-starter feed. The chicks were distributed in the transportation boxes in a completely randomized design in five treatments and five replicates of 60 birds each.

The product was added “over the top” replacing starch in a basal pre-starter feed (Table 1) formulated with corn and soybean meal, following the Rostagno et al. (2011) nutritional recommendations.

Table 1
Composition and values calculated (%) for the basal diet.

After hatch, neonates were vaccinated and sexed, following the standard proceedings of the hatchery, and then they were transported to the aviary. All proceedings conducted with the chicks, from hatching until housing, lasted 24 hours.

During this 24-hour period, the chicks received 3 g/bird of pre-starter feed and did not receive water, in order to simulate what happens in the hatchery. After 24 hours of experimental feed intake, ten chicks per replicate (50 chicks per treatment), according to the average plot weight, were transferred to galvanized steel cages equipped with linear feeders and water dispensers. The chicks continued to receive the same experimental feeds in the cages until they reached seven days of age.

Water and feed were offered ad libitum over the whole rearing period, with a 24-hour light cycle. The maximum and minimum environmental temperatures were monitored twice a day (8:00 AM and 5:00 PM) using thermometers within the shed. The mean maximum and minimum temperature during the experiment were 33.47°C and 25.48°C, respectively.

Before the chicks went to the transportation box, 15 chicks were separated as controls and the data on average body weight 44.54g, average yolk sac weight 5.87g, average intestine weight 1.87g and blood glucose 348.2 were collected. After 24 hours in the transportation box, five chicks per treatment were chosen at random. They were evaluated by body weighing, weighing of the yolk sac and intestine, collection of fragments from the duodenum, jejunum and ileum, and measurement of glucose concentration. The rest of the chicks were housed in the batteries according to the treatments. At seven days of age, another five broilers per treatment were separated and body weight and relative weight of yolk sac and total intestine, collected from the duodenum jejunum and ileum fragments for histomorphometric measurements and blood samples for glucose concentration determination were evaluated.

For performance evaluation at seven days of age, weight gain was calculated as the difference between the final and initial weights. Feed intake was calculated as the difference between the feed supplied and the leftovers at the end of each phase. Feed conversion was obtained through the ratio of feed intake/weight gain. Livability was assessed through the formula V (%) = 100 - mortality.

Glucose concentration was assessed through analysis within thirty seconds after drawing blood, using portable glucose metering equipment (Accu-Chek Active Roche®)

Five chicks from each treatment were euthanized to evaluate intestinal development and yolk sac retraction. The yolk sac and intestines (small and large) were then weighed on a scale precision of 0.0001 g.

One chick per replicate was sampled for histomorphometric measurements of the small intestine. Two-centimeter fragments were collected from the duodenum, jejunum and ileum, and were fixed in buffered 10% formaldehyde for 24 hours. These samples were then subjected to standard histological procedures to produce histological slides. The sections were stained with hematoxylin and eosin (Hu et al., 2012Hu CH, Gu LY, Luan ZS, Song J, Zhu K. Effects of montmorillonite-zinc oxide hybrid on performance, diarrhea, intestinal per­meability and morphology of weanling pigs. Animal Feed Science and Technology 2012;177(1-2):108-115.; Sousa et al., 2015Sousa DC, Oliveira NL, Santos ET, Guzzi A, Dourado LR, Ferreira GJ. Caracterização morfológica do trato gastrointestinal de frangos de corte da li­nhagem Cobb 500(r). Pesquisa Veterinária Brasileira 2015;35(Supl 1):61-68.). The heights of ten villi and the depths of ten crypts were measured in each section.

A metabolic trial was conducted between seven and ten days of age, using the total excreta collection method. Excreta were collected twice a day (8:00 AM and 5:00 PM), stored in plastic bags (which were identified per replicate) and kept in a freezer. At the end of the experimental period, the amount of feed intake and excreta produced were determined. The samples of excreta and experimental feed were used to determine the dry matter, nitrogen and ether extract content, as described by Silva & Queiroz (2002Silva DJ, Queiroz AC. Análise de alimentos: métodos químicos e biológicos. 3rd ed. Viçosa: Editora UFV; 2002.).

The results from bromatological analyses were used to calculate coefficients of metabolizability (MC) for dry matter (MCDM), nitrogen (MCN) and ether extract (MCEE). Nitrogen balance (NB) and ether extract balance (EEB) were also calculated by Sakomura & Rostagno (2007Sakomura NK, Rostagno HS. Métodos de pesquisa em nutrição de monogástricos. Jaboticabal: FUNEP; 2007.).

All data analysis was evaluated for normality by the Shapiro-Wilk test. Polynomial regression analysis was performed concerning betaine concentrations tested (p<0.05).

RESULTS AND DISCUSSION

When mean live weights of the chicks before and 24 hours after the experimental feed and the amount of feed intake were analyzed (Table 2), it was observed that the birds did not consume a significant amount of feed and therefore experienced mean weight loss of 2.52 g over the 24 hours inside the transportation boxes. The fact that the chicks did not have access to water during this period may contribute to weight loss. The reduction of live weight of the chicks during this phase occurs due to higher use of energy reserves from the yolk sac, digestive and renal excretions, and dehydration (Leu et al., 2002Leu WMK, Cotta JTB, Oliveira AIG, Rodrigues PB. Desempenho de frangos submetidos à restrição alimentar na fase Zinicial em diferentes sistemas de criação. Ciência e Agrotecnologia 2002;26(3):610-617.; Carvalho et al., 2013Carvalho LSS, Machado CA, Fagundes NS, Litz FH, de Abreu Fernandes E. Desenvolvimento biométrico e desempenho de frangos de corte submetidos a diferentes períodos de jejum pós-eclosão. Brazilian Journal of Veterinary Research and Animal Science 2013;50(4):300-306.). The treatments in this experiment were not sufficient to reduce these effects (p>0,05).

Table 2
Mean weight loss and feed intake of pre-starter diet supplemented with betaine of newly hatched chicks, before and after a 24-hour waiting period in transportation boxes.

The live weight of the chicks, retraction of the yolk sac, small intestine weight and glucose levels (Table 3) were not affected by betaine supplementation, probably due to the low feed intake in the transportation box. Different results were observed by Bhanja et al. (2009Bhanja SK, Anjali Devi C, Panda AK, Shyam Sunder G. Effect of post-hatch feed deprivation on yolk-sac utilization and performance of young broiler chickens. Asian Australasian Journal Animal Science 2009;2(2):1174-1179.), who verified more significant reduction of yolk sacs in birds fed immediately after hatch, thus suggesting that feed intake might increase the mechanical activity of the intestine, such that the yolk sac would be absorbed more rapidly.

Fasting chicks go through a critical gluconeogenic metabolism period, with increases in ketosis and dehydration. Quick feed supplying could minimize these effects by altering the glycemic pattern of these birds (Reis, 2018Reis TL. Early nutrition of broiler chicks. Ciência Animal 2018;28(1):82-97.). Gluconeogenesis decreases as the plasma glucose concentrations increase through feed intake. Our results showed that chickens are relatively hyperglycemic compared to mammals. Hyperglycemia is independent of feed intake. In the experiment conducted by Zhao et al. (2014Zhao X, Sumners LH, Gilbert ER, Siegel PB, Zhang W, Cline M. Delayed feeding after hatch caused compensatory increases in blood glucose concentration in fed chicks from low but not high body weight lines. Poultry Science 2014;93:617-624.), the authors observed blood concentrations higher than 200 mg/dL at hatch (before feed intake).

Table 3
Evaluation of yolk sac retraction, intestine weight and blood glucose of chicks fed a pre-starter diet supplemented with betaine, after a 24-hour waiting period in transportation boxes.

(Table 4) presents the results regarding the development of intestinal villi in the chicks fed pre-starter diet supplemented with betaine. Betaine supplementation influenced linearly (p=0,027) the ileum crypt depth (Y=32.6342 + 0.0401451X, R²=0.85).

Table 4
Evaluation of the villi development (villi height, crypt depth and villus/crypt ratio) in the intestine of chicks fed a pre-starter diet supplemented with betaine after a 24-hour waiting period in transportation boxes.

Increasing of crypt depth implies higher rates of cells proliferation and renovation of the intestinal epithelium, thereby increasing the number of villi since these structures originate from the lower part of the crypt (Fernandes et al., 2017Fernandes JIM, Rorig A, Gottardo ET, Schmidt JM, Burin Júnior AM, Fulber LM. Post-hatch diets supplemented with sources of fat and added taurine and glycine on the intestinal morphology and performance of broilers at 1 to 21 days. Arquivo Brasileiro de Medicina Veterinária e Zootecnia 2017;69(1):198-204.). The lumen of the intestine is always hyperosmotic when compared to blood plasma, so osmoregulatory substances are beneficial for intestinal cells (Mongin, 1976Mongin P. Ionic constituents and osmolality of the small intestinal fluids of the laying hen. British Poultry Science 1976;17:383-392.). Betaine has the function of controlling osmotic pressure in intestinal cells (Kettunen et al., 2001Kettunen H, Peuranen S, Tiihonen K. Betaine aids in the osmoregulation of duodenal epithelium of broiler chicks, and affects the movement of water across the small intestinal epithelium in vitro. Comparative Biochemistry and Physiology Part A: Molecular & Integrative Physiology 2001;129:595-603.). This osmoprotective function must have helped the linear increase in the depth of the Ileum crypt with betaine supplementation.

Chick feed intake was altered by betaine supplementation in the pre-starter diet (Table 5). There was a quadratic effect, with an increase in feed intake until a supplementation of 0.152% betaine was reached (Y = 168.242 + 0.12448X - 0.409295x², R²= 0.62), from that value the feed intake was reduced, but did not interfere in weight at seven days. A similar result was found by Teixeira et al. (2006Teixeira M, Niang TMS, Gomes AVC, Lopes CWG. Efeito do uso da betaína na biologia e morfologia dos estádios evolutivos de Eimeria acervulina em frangos de corte infectados experimentalmente com oocistos esporulados. Revista Brasileira de Parasitologia Veterinária 2006;15(4):193-198.), studying levels of betaine inclusion (0.00; 0.05; 0.10 and 0.15%), who reported that birds that received 0.10% betaine had lower feed intake at seven days of age. However, this reduction in consumption did not affect weight gain. However, betaine supplementation in the feed did not interfere with the chick’s feed intake at 7 days (Pereira et al., 2010Pereira PWZ, Menten JFM, Racanicci AMC, Traldi AB, Silva CS, Rizzo PV. Avaliação de complexo enzimático e betaína natural em rações para frangos de corte criados em aviário comercial. Revista Brasileira de Zootecnia 2010;39(10):2230-2236.), but improved the final weight and conversion at 14 days (El-Shinnawy, 2015El-Shinnawy AM. Effect of betaine supplementation to methionine adequate diet on growth performance, carcass characteristics, some blood parameters and economic efficiency of broilers. Journal of Animal and Poultry Production 2015;6(1):27-41.) and feed conversion at 21 days (Amerah & Ravindran, 2015Amerah AM, Ravindran V. Effect of coccidia challenge and natural betaine supplementation on performance, nutrient utilization, and intestinal lesion scores of broiler chickens fed suboptimal level of dietary methionine. Poultry Science 2015;94(4):673-680.).

Table 5
Performance in the pre-starter phase (1 to 7 days) among chicks that fed diets supplemented with betaine.

The current experiment evaluated only the pre-starter phase. Therefore, a considerable amount of betaine may be preferentially applied by enterocytes to stimulate intestinal development and function in chicks rather than improve growth performance during the starter period. Thus, these effects may affect the overall performance of broilers in the future.

Betaine supplementation did not influence the retraction of the yolk sac, the absolute intestine weight and the plasma glucose concentration at seven days of age (Table 6). During the first-week after hatching, the intestinal weight of these chicks increases dramatically when compared to the total body weight (Uni et al., 2003Uni Z, Tako E, Gal-Garber O, Sklan D. Morphological, molecular and functional changes in the chicken small intestine of the late-term embryo, Poultry Science 2003;82:1747-1754.). Therefore, the effects of substances beneficial to the development of the intestine are more prominent in the first-week of the broiler. Tissues that rely on betaine as an osmolyte include the leukocytes, kidney, liver, brain and intestines (Klasing et al., 2002Klasing KC, Adler KI, Remus JC, Calvert CC. Dietary betaine increases intraepithelial lymphocytes in the duodenum of coccidian-infected chicks and increases functional properties of phagocytes. The Journal of Nutrition 2002;132:2274-2282.). In the current study, there was no effect of betaine supplementation on intestinal weight. Regardless of the treatments, the results showed that the intestine increased during the first-week post-hatch, whereas the yolk sac weight decreased rapidly, which is indicative of the maturation of digestive.

Even with stress in the transportation box and reduced feed intake, glucose concentration in chicks at seven days was also not influenced by betaine supplementation. The negative effect of stress on the central nervous system can reduce the metabolic rate and feed intake. This stress reaction process generates higher energy expenditure by the body and osmotic imbalance in the cells. Therefore, betaine supplementation can reestablish the osmotic balance in cells, mitigating the deleterious effects of stress (Klasing et al. 2002Klasing KC, Adler KI, Remus JC, Calvert CC. Dietary betaine increases intraepithelial lymphocytes in the duodenum of coccidian-infected chicks and increases functional properties of phagocytes. The Journal of Nutrition 2002;132:2274-2282.). Layers created in heat stress showed a higher concentration of glucose when supplemented with 0.10% betaine (220 mg / dL) compared to the control (208 mg / dL) (Attia et al. 2016Attia YA, Aelh EH, Abedalla AA, Berika MA, Alharthi MA, Kucuk O, et al. Laying performance, digestibility and plasma hormones in laying hens exposed to chronic heat stress as affected by betaine, vitamin C, and/or vitamin E supplementation. SpringerPlus 2016;5:1619-1631.). Contrary to these results, Konca et al. (2008Konca Y, Kirkpinar F, Mert S, Yayalak E. Effects of betaine on performance, carcass, bone and blood characteristics of broiler during natural summer temperatures. Journal of Animal and Veterinary Advances 2008;7(8):930- 937.), Sayed & Downing (2011Sayed MAM, Downing J. The ef-fects of water replacement by oral rehydration fluids with or without betaine supplementation on performance, acid-base balance, and water retention of heat-stressed broiler chickens. Poultry Science 2011;90:157-167.) and El-Shinnawy (2015El-Shinnawy AM. Effect of betaine supplementation to methionine adequate diet on growth performance, carcass characteristics, some blood parameters and economic efficiency of broilers. Journal of Animal and Poultry Production 2015;6(1):27-41.) evaluated betaine supplementation in the diet of broilers and found no difference in serum glucose levels.

Table 6
Evaluation of yolk sac retraction and intestinal growth among chicks that were given pre-starter feed supplemented with betaine, at the age of seven days.

Betaine supplementation influenced jejunum villus/crypt ratio (Table 7), presenting quadratic effect (p=0.049) with minimum point in 0.087% of supplementation (Y=5.20261 - 0.0074454X + 0.00425619X², R²=0.74). This fact demonstrated an improvement in the integrity of the jejunum with supplementation above 0.1% betaine.

Table 7
Evaluation of intestinal villi development in chicks fed a pre-starter diets supplemented with betaine at seven days of age.

In pigs, betaine decreased the energy expenditure of intestinal cells due to its osmoregulatory effect (Siljander-Rasi et al., 2003Siljander-Rasi H, Peuranen S, Tiihonen K, Virtanen E, Kettunen H, Alaviuhkola T, et al. Effect of equi-molar dietary betaine and choline addition on performance, carcass quality and physiological parameters of pigs. Animal Science Journal 2003;76:55-62.). The most significant effects of betaine have been reported when broilers are challenged by pathogens that affect villi, as shown by Klasing et al. (2002Klasing KC, Adler KI, Remus JC, Calvert CC. Dietary betaine increases intraepithelial lymphocytes in the duodenum of coccidian-infected chicks and increases functional properties of phagocytes. The Journal of Nutrition 2002;132:2274-2282.), who after seven days of inoculating broilers with Eimeira, found that those fed 0.10% betaine presented higher villi.

The results from the metabolic trial demonstrated that there were no differences in betaine concentrations, except for the ether extract metabolizability coefficient (Table 8) that was improved until reaching 0.163% supplementation (Y= 54.3717 + 200.9654X - 671.5212X², R²= 0.47).

Table 8
Nitrogen balance (NB), ether extract balance (EEB), nitrogen metabolizability coefficient (NMC), ether extract metabolizability coefficient (EEMC) and dry matter metabolizability coefficient (DMMC) of chicks fed a pre-starter diet supplemented with betaine, from seven to ten days of age.

Betaine has the function of donating methyl groups mainly for homocysteine that will provide the formation of methionine, therefore it is involved in protein and energy metabolism (Eklund et al., 2005Eklund M, Bauer E, Wamatu J, Mosenthin R. Potential nutritional and physiological functions of betaine in livestock. Nutrition Research Reviews 2005;18:31-48.). In broiler diets with methionine deficiency, betaine supplementation improved the metabolizability of ether extract and protein (El-Husseiny et al., 2007El-Husseiny OM, Abo-El-Ella MA, Abd-Elsamee MO, Ab-Elfattah MM. Response of broilers performance to dietary betaine and folic acid at different methionine levels. International Journal of Poultry Science 2007;6:515-523.).

Betaine for being osmoregulatory helps to maintain the integrity of the intestinal epithelium, which is important for improving the absorption and digestibility of nutrients. Betaine supplementation has shown an improvement in the use of some nutrients in birds of different ages. Amerah & Ravindran (2015Amerah AM, Ravindran V. Effect of coccidia challenge and natural betaine supplementation on performance, nutrient utilization, and intestinal lesion scores of broiler chickens fed suboptimal level of dietary methionine. Poultry Science 2015;94(4):673-680.) with supplementation of 0.096% betaine for 21-day-old chickens found better digestibility of starch, ash and ten amino acids; Ratriyanto et al. (2014Ratriyanto A, Indreswari R, Sunarto S. Effects of protein levels and supplementation of methyl group donor on nutrient digestibility and performance of broiler chickens in the tropics. International Journal of Poultry Science 2014;13:575-581.), with 0.14% inclusion of betaine found a higher percentage of dry matter, protein and crude fiber in chickens aged 39 days and Attia et al. (2016Attia YA, Aelh EH, Abedalla AA, Berika MA, Alharthi MA, Kucuk O, et al. Laying performance, digestibility and plasma hormones in laying hens exposed to chronic heat stress as affected by betaine, vitamin C, and/or vitamin E supplementation. SpringerPlus 2016;5:1619-1631.) challenged layers to heat stress and observed improvement in utilization of protein with betaine supplementation.

In the current study, this osmoprotective effect was not directly evaluated, but there was an improvement in the use of the ether extract. Possibly, chicks that were fed diets containing betaine were favored by the ability of this substance to act as an osmolyte, maintaining the integrity of the intestinal mucosa, improving digestibility and the absorption capacity of nutrients.

CONCLUSIONS

Betaine supplementation around 0.150% increases feed intake, the use of ether extract and interferes with the intestinal villi of chicks at seven days of age.

REFERENCES

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Publication Dates

  • Publication in this collection
    18 Dec 2020
  • Date of issue
    2020

History

  • Received
    04 June 2020
  • Accepted
    05 Sept 2020
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