1. Introduction
Recent advances in plant breeding increase differences in the chemical composition of grain between varieties within each cereal species [
1,
2]. In the case of wheat and maize grain, which are most often used in poultry nutrition, individual variability may differ not only in protein, starch, or fiber content but also in nutrient digestibility and metabolizable energy content [
3]. Unfortunately, this fact is not widely considered during diet formulation for animals and very small varietal differences are assumed.
A large number of new cereal varieties makes it necessary to develop simple procedures for determining their nutritional value without time-consuming and expensive studies on animals. This opportunity is provided by regression equations used for estimating digestible nutrient and metabolizable energy content based on laboratory data. The improvement of analytical procedures has contributed to the elaboration of rapid and efficient methods for the determination of nutrients, in particular the cell wall content of plant feeds [
4]. The value of metabolizable energy in cereal grain is negatively correlated to crude fiber content; however, it is believed that this kind of fiber determination is not sufficiently described within the energy value for mixtures containing cereal grain [
5,
6]. In this context, the effect of crude, detergent, or dietary fiber content, including soluble and insoluble fractions, on energy utilization in poultry, has not been adequately studied.
The equations for estimating the energy value of feeds for poultry are based on digestible nutrient content and fail to account for unique differences of carbohydrate structure between cereal varieties; however, some research [
7,
8,
9] indicates a significant role of cell wall structural components in the estimation of the apparent metabolizable energy (AME
N) value of feeds. To more accurately predict the nutritive effect of fiber from raw materials, a better characterization of fiber fractions, their degradation in the chicken, and their physiological effects are required. Therefore, we hypothesized that the inclusion of different fiber fraction content into regression equations may significantly improve the efficiency and accuracy of metabolizable energy prediction. Additionally, replacing data related to digestible nutrients with crude nutrients, especially with crude, detergent, or dietary fiber content, may facilitate the prediction procedure. The study aimed to determine the effect of nutrient content, with special emphasis on carbohydrates, on nutrient digestibility, nitrogen balance, and apparent metabolizable energy content (AME
N) of wheat and maize grain for broiler chickens.
2. Materials and Methods
The study was conducted at the Department of Animal Nutrition and Biotechnology and Fisheries of the University of Agriculture in Krakow (Poland). Grain from 6 varieties of spring wheat, 3 varieties of winter wheat, and 9 varieties of maize (flint, semi flint, semi dent, and dent) were selected and prepared (dried and ground) for further procedures (
Table 1). Plants were grown in the same year in the experimental farm Małopolska Plant Raising HBP Ltd. near Kraków located in south-eastern Poland. The crops were grown on Haplic Phaeozem formed from loess and classified as very good wheat complex soil. The physicochemical properties of the soil were appropriate for the habitat requirements of both species. For each wheat and maize variety, chemical analysis of grain were performed in 4 different representative samples originating from 4 different experimental crop plots.
The in vivo digestibility and balance trials were carried out separately for each variety of wheat and maize grain with 90 Ross 308 chickens, 42 to 49 days of age (2 × 9 groups with 5 birds per group in 2 replicates each; n = 80 for wheat and maize grain, separately). Before the digestibility trials, chickens were kept in pens and fed first the standard broiler starter diet (from 0 to 14 d of life) and then (from 14 to 42 d of life) they were kept in individual cages and fed diets based on the evaluated wheat or maize cultivars and containing (g/kg):
wheat 732.6, soybean meal 150, fish meal 80, monocalcium phosphate 5.8, limestone 18, sodium chloride 3.0, L-lysine 2.6, DL-methionine 3.0, and vitamin-mineral premix 5;
maize 708.5, soybean meal 180, fish meal 80, monocalcium phosphate 5, limestone 11, L-lysine 4.7, DL-methionine 2.8, vitamin-mineral premix 5, to satisfy nutrient requirements of broilers.
Nutritive values of mixtures within each cereal species were similar and contained an average 11.31 and 10.88 AMEN, 20.2% and 20.8% CP, and 1.19% and 1.18% Lys, respectively, for wheat and maize.
During the digestibility trials, chickens were kept individually in metabolic cages with free access to water. Grain was fed ad libitum as coarsely ground meal which could pass through a 4.0 mm screen sieve in 2000 rpm speed in hammer mill. Both pre-treatment and data collection periods lasted 4 days (in total, 8 days per trial). Excreta were collected twice a day from trays placed under each cage and stored at −18 °C. All methods and procedures in this study were approved and followed the recommendations of the Local Ethics Committee in Krakow (Poland).
Prior to chemical analysis, air-dried samples of wheat and maize grain were ground to pass through a 1 mm sieve with a Pulverisette 15 Laboratory Cutting Mill (Fritsh GMBH, Idar-Oberstein, Germany) and analyzed for content of dry matter (DM), ash, crude protein (CP), ether extracts (EE), and crude fiber (CF) using standard analytical procedures (procedure nos. 934.01, 942.05, 976.05, 920.39, and 962.09, for DM, ash, CP, EE, and CF, respectively; [
10]. Neutral detergent fiber determined with heat-stable amylase (aNDF) [
11], acid detergent fiber (ADF) [
10]; official method 973.18 and acid detergent lignin (ADL) [
4] were determined using an Ankom220 Fiber Analyzer (Ankom Technology, NY, USA). The starch content was determined by an enzymatic method [
12]. The same procedures were used for chemical analyses of the excreta. Gross energy (GE) content was determined using a bomb calorimeter (KL-10, PRECYZJA, Bydgoszcz, Poland). The content of dietary fiber (soluble—SDF and insoluble—IDF) was determined based on 991.43 AOAC [
10] procedure [
13]. Water-soluble carbohydrates were analyzed spectrophotometrically using the color reaction with anthrone in concentrated, purified H
2SO
4. Prior to measurements, the samples were deproteinized using zinc acetate Zn(CHCOO)
2·H
2O 275.12 g/L
−1 water and potassium ferrocyanide K
4Fe(CN
6) 3H
2O 171.99 g/L
−1 water. Extinction was measured at a wavelength of λ = 620 nm [
14]. The proportion of amylose and amylopectin in starch was determined following the method described by Morrison et al. [
15].
The results related to the chemical composition of feeds and excreta were used to calculate the coefficients of dry matter, organic matter, ether extracts, and N-free extract apparent digestibility. The apparent crude protein digestibility was calculated using the alpha-amino nitrogen (N-α-NH
2) method [
16] modified by Barteczko et al. [
17]. This method is based on the determination of alfa-amino groups in the feces from undigested feed proteins. The first stage is the hydrolysis of feed protein and feces in hydrochloric acid (HCl). Subsequently, the feces samples are subjected to pressure distillation to remove the non-protein nitrogen fraction (mainly ammonia-NH
3), which can positively react with ninhydrin and thus affect the results. Hydrolysates of the distillation feces and the corresponding feed have undergone a reaction during which free amino groups formed a colored complex with ninhydrin. Next, the extinction of fecal samples and corresponding feed samples were measured at the wavelength of 570 nm.
Nitrogen balance (BN), called also as “N retained”, was calculated according to the formula:
The metabolizable energy value corrected to zero nitrogen balance (i.e., to the nitrogen equilibrium of the birds; AME
N) was calculated according to the formula:
The regression equations for the provision of AME
N content were estimated using multiple forward stepwise regression [
18]. The significance of model parameters was analyzed using Student’s t-test. The coefficient of determination (R
2), the standard deviation of the difference between actual and estimated values (RSD), and mean estimation error (S) were also considered. The results of chemical analysis and digestibility trials were analyzed statistically by one-way ANOVA and Tukey’s range test [
18]. The main experimental factor for wheat grains included the differences between winter and spring varieties while for maize grains included the differences between early, medium early, and medium late forms of varieties. The significance level was set at
p ≤ 0.05. Tendencies were discussed at 0.05 <
p < 0.10 unless otherwise stated. All data are reported as least squares means with a pooled standard error of the means.