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. 2023 Dec 18;103(3):103382. doi: 10.1016/j.psj.2023.103382

Effects of dietary methionine supplementation on the growth performance, immune responses, antioxidant capacity, and subsequent development of layer chicks

Jiatu Zhang *,†,1, Shunju Geng *,1, Yahao Zhu *, Lan Li *, Lihong Zhao *,†, Qiugang Ma ⁎,†, Shimeng Huang ⁎,†,2
PMCID: PMC10792981  PMID: 38176373

Abstract

Deficiencies or excesses of dietary amino acids, and especially of methionine (Met), in laying hens can lead to abnormal protein anabolism and oxidative stress, which affect methylation and cause cellular dysfunction. This study investigated the effects of dietary methionine (Met) levels on growth performance, metabolism, immune response, antioxidant capacity, and the subsequent development of laying hens. A total of 384 healthy 1-day-old Hyline Grey chicks of similar body weight were randomly allocated to be fed diets containing 0.31%, 0.38%, 0.43% (control group), or 0.54% Met for 6 wk, with 6 replicates of 16 chicks in each. The growth performance of the chicks was then followed until 20 wk old. The results showed dietary supplementation with 0.43% or 0.54% Met significantly increased their mean daily body weight gain, final weight, and Met intake. However, the feed:gain (F/G) decreased linearly with increasing Met supplementation, from 0.31 to 0.54% Met. Met supplementation increased the serum albumin, IgM, and total glutathione concentrations of 14-day-old chicks. In contrast, the serum alkaline phosphatase activity and hydroxyl radical concentration tended to decrease with increasing Met supplementation. In addition, the highest serum concentrations of IL-10, T-SOD, and GSH-PX were in the 0.54% Met-fed group. At 42 d of age, the serum ALB, IL-10, T-SOD, GSH-PX, T-AOC, and T-GSH were correlated with dietary Met levels. Finally, Met supplementation reduced the serum concentrations of ALP, IL-1β, IgA, IgG, hydrogen peroxide, and hydroxyl radicals. Thus, the inclusion of 0.43% or 0.54% Met in the diet helps chicks achieve superior performance during the brooding period and subsequently. In conclusion, Met doses of 0.43 to 0.54% could enhance the growth performance, protein utilization efficiency, antioxidant capacity, and immune responses of layer chicks, and to promote more desirable subsequent development during the brooding period.

Key words: Met, layer chick, growth performance, antioxidant capacity, serum biochemistry

INTRODUCTION

Throughout the life cycles of both humans and animals, the early years are crucial (Lilburn and Loeffler, 2015; Taha-Abdelaziz et al., 2018; Liu et al., 2022; Xu et al., 2022). Importantly, early nutrition is closely associated with health and physical status in the later stages of life (Ratsika et al., 2021; Abdel-Moneim et al., 2022; Haiden and Haschke, 2023; Jin et al., 2023). Amino acids play important roles in the regulation of food intake and nutrient metabolism (Wu, 2009), as well as being the building blocks of proteins (Wu, 2010; Wu et al., 2014). Given that they are involved in nearly all cellular functions, amino acids are essential to life (Wu et al., 2014; Li et al., 2023), and they are therefore key components of human and animal diets, in the forms of dietary protein or supplements.

Sulfur-containing amino acids (SAAs) are functional amino acids that affect protein structure, metabolism, oxidative stress status, and immune responses (Grimble, 2006; Li et al., 2021). They include methionine (Met) and cysteine, which have important roles in animal health and development (Martínez et al., 2017; Colovic et al., 2018; Elango, 2020b; Teng et al., 2023). In addition, a deficiency of Met has negative effects on the growth performance, carcass traits, meat quality, and oxidative status of breast muscle in poultry (Sekiz et al., 1975; Wen et al., 2017; Barekatain and Kluenemann, 2023). Previous studies have shown that dietary Met levels higher than those recommended by the National Research Council (NRC) in 1984 improve weight gain, feed conversion efficiency (Wen et al., 2017), and growth performance of broilers throughout rearing (Majdeddin et al., 2019). However, the effects of appropriate dietary Met levels during the brooding period on the growth performance, antioxidant capacity, and development of layer chicks have not been fully described.

Sulfur-containing amino acids are amino acids that contain sulfhydryl groups and play crucial roles in protein structure, metabolism, immunity, and oxidation (Brosnan and Brosnan, 2006; Stipanuk, 2020). In corn–soybean diets, Met is an important limiting SAA (Stipanuk, 2020). It plays an active role in protein synthesis, immune function, antioxidant activity (Elango, 2020b; Teng et al., 2023), and skeletal development (Castro et al., 2020). However, imbalances in diets containing Met may alter feed intake and/or cause defects in digestion and absorption (Fagundes et al., 2020). When Met is introduced into broiler diets in appropriate amounts, it reduces oxidative stress and improves protein utilization, resulting in lower production costs (Ullrich et al., 2019; Millecam et al., 2021). Interestingly, a dietary Met level higher than that required for growth performance improves the antioxidant status in liver and muscle of broilers exposed to heat stress (Zeitz et al., 2020). Conversely, a large number of previous studies have shown that Met deficiency has adverse effects on the growth performance, amino acid metabolism, and intestinal development of laying hens, and may affect their subsequent development during the growing period (Liu et al., 2022). Therefore, supplementation of the diets of laying hens with an appropriate level of Met during their early life has significant effects on their subsequent production. However, few studies have evaluated the effects of a range of levels of dietary Met supplementation on the subsequent growth and development of layer chicks. In the current study, the focus was on exploring the impact of dietary Met supplementation on the growth performance and antioxidant capacity in chicks, as well as its influence on their subsequent development.

MATERIALS AND METHODS

Animal Ethics

The study was approved by the Animal Protection and Utilization Committee of China Agricultural University (AW82202202-1-1, Beijing, China), and was conducted in accordance with the Guidelines for the Use of Experimental Animals of the Ministry of Science and Technology (Beijing, China).

Birds, Diets, and Management

A total of 384 healthy 1-day-old Hyline Grey chicks with similar body weight (BW) were provided by Beijing Deqingyuan Agriculture Technology Co. Ltd. (Beijing, China) and randomly allocated to 4 treatment groups, with 6 replicates and 16 birds per replicate. The groups were fed pelleted diets containing 0.31%, 0.38%, 0.43% (control group), or 0.54% Met (measured values) from 1-day-old until they were 6 wk old. The birds were housed in stainless steel chicken cages (37 × 65 × 62 cm) that were equipped with nipple drinkers and external access devices for feed and water throughout the experimental period. During rearing, the room temperature, humidity, and artificial light provision were strictly controlled, and a vaccination program was followed, according to the management guidelines for Hyline Grey chicks. At the end of the diet feeding period, the birds were transferred to the standard diet and followed until they were 20 wk old. The diets were formulated with the nutrient contents recommended by the NRC in 2012 (Table 1).

Table 1.

The composition and nutrient levels of diets.

Items Met level/%
0.31 0.38 0.43 0.54
Ingredients
Corn 60.44 60.44 60.44 60.44
Soybean meal 32.10 32.10 32.10 32.10
Limestone 1.20 1.20 1.20 1.20
CaHPO4 1.70 1.70 1.70 1.70
NaCl 0.30 0.30 0.30 0.30
Vitamin premixa 0.04 0.04 0.04 0.04
Trace element premixb 0.30 0.30 0.30 0.30
DL-Methionine - 0.09 0.18 0.27
Lysine (65%) 0.11 0.11 0.11 0.11
Threonine 0.03 0.03 0.03 0.03
Choline chloride 0.10 0.10 0.10 0.10
Soybean oil 2.30 2.30 2.30 2.30
Zeolite powder 1.38 1.29 1.20 1.11
Total 100.00 100.00 100.00 100.00
Nutrient levels
Crude protein (%) 18.86 18.95 19.04 19.13
ME/(MJ/kg) 12.18 12.18 12.18 12.18
Ca (%) 0.94 0.94 0.94 0.94
Total P (%) 0.66 0.66 0.66 0.66
Non-phytate phosphorus (%) 0.41 0.41 0.41 0.41
Met (%) 0.31 (0.31) 0.4 (0.38) 0.49 (0.43) 0.58 (0.54)
Met + Cys c (%) 0.67 (0.66) 0.76 (0.74) 0.85 (0.79) 0.94 (0.91)
Lys (%) 1.09 1.09 1.09 1.09
Try (%) 0.22 0.22 0.22 0.22
Thr (%) 0.72 0.72 0.72 0.72
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a

Vitamin premix provided the following per kg of diet: vitamin A, 8,000 IU; vitamin D3, 3,600 IU, vitamin E, 21 IU, vitamin B1, 3 mg, vitamin B2, 10.2 mg, vitamin K3, 4.2 mg, vitamin B6, 5.4mg, vitamin B12, 0.024 mg, calcium pantothenate, 15 mg, folic acid, 0.9 mg, biotin, 0.15 mg, nicotinic acid, 45 mg.

b

Trace element premix provided the following per kg of diet: Zn, 83 mg, Fe, 66 mg, Mn, 80 mg, Cu, 6.8 mg, Se, 0.3 mg, I, 1 mg.

c

The dietary SAA levels were calculated values, and their analyzed values were in brackets. 0.43% Met as the control group.

Growth Performance

The BW of each chick at 1-day-old was recorded as the initial body weight (IBW) and the feed intake of each was calculated at the end of each week. The chicks were weighed after fasting at the end of the 2nd, 4th, and 6th weeks of the feeding period, and their average daily gain (ADG), average daily feed intake (ADFI), feed:gain (F/G), and tibial length (TL) were recorded. Further measurements were then made from 6 wk old until 20 wk old: BM and TL were measured when the chicks were 8, 10, 12, 14, 16, 18, and 20 wk old.

Blood Collection and Analysis

Blood samples were collected after 2 and 6 wk of diet-feeding. Two birds were randomly selected from each replicate group for blood sampling from a wing vein, then the samples were centrifuged at 3,000 rpm for 15 min, and serum samples were placed into 1.5-mL centrifuge tubes and stored at −20°C (Wang et al., 2019). The serum concentrations of urea, uric acid (UA), total protein (TP), albumin (ALB), and glucose (GLU), and the activities of lactate dehydrogenase (LD) and alkaline phosphatase (ALP) were measured using a Hitachi 7600 automatic serum biochemistry analyzer (Tokyo, Japan). The serum globulin (GLB) concentration and the albumin-to-globulin ratio (A/G) were calculated using the total protein and albumin concentrations, according to the instructions provided by the manufacturer. In addition, the serum concentrations/activities of interleukin 1β (IL-1β), interleukin 6 (IL-6), tumor necrosis factor α (TNF-α), interleukin 10 (IL-10), immunoglobulin A (IgA), immunoglobulin M (IgM), immunoglobulin G (IgG), total superoxide dismutase (T-SOD), malondialdehyde (MDA), glutathione peroxidase (GSH-PX), catalase (CAT), total glutathione (T-GSH), total antioxidant capacity (T-AOC), hydrogen peroxide (H2O2), and hydroxyl radicals (·OH) were measured using the corresponding kits (Nanjing Jiancheng, Nanjing, China).

Statistical Analysis

The data are expressed as mean and standard error of mean and were analyzed using general linear model analysis and Duncan's multiple comparison method in SPSS v.18.0 (IBM, Inc., Armonk, NY). SAS v.8.0 (Cary, NC) was used for linear and quadratic comparative analyses of the effects of dietary Met level on the birds during the brooding stage and subsequently, and quadratic curves or fold lines were fitted to the data with significant regularity during the analysis. P < 0.05 was regarded as indicating statistical significance.

RESULTS

Effect of Dietary Met Level on the Growth Performance of Hyline Grey Chicks

As shown in Table 2, there were significant effects of Met intake on ADG, F/G, and FBW. Increasing Met in the brood diet significantly increased ADG and FBW (P < 0.05), and F/G decreased linearly with increasing dietary Met content (P < 0.05). The ADG and FBW were significantly higher in the 0.43% and 0.54% Met groups than in the 0.31% Met group (P < 0.05), and 0.43 to 0.54% Met was associated with lower F/G and the most uniform body weight at 6 wk old versus the control group (0.31% Met).

Table 2.

Effect of dietary Met levels on growth performance of chicks at 42 d of age.

Items 0.31% Met 0.38% Met 0.43% Met 0.54% Met SEM P-value Linear Quadratic
IBW (g) 41.91ab 41.81b 42.01a 41.87ab 0.049 0.046 0.678 0.65
ADFI (g) 21.47 22.43 22.15 22.72 0.61 0.527 0.22 0.76
ADG (g) 9.08c 9.86b 10.24ab 10.62a 0.14 <0.001 < 0.001 0.16
F/G 2.31a 2.24ab 2.15b 2.12b 0.04 0.016 0.002 0.68
FBW (g) 423.26c 455.94b 472.29ab 488.97a 5.82 <0.001 <0.001 0.18
SAA intake (g/d) 0.14d 0.17c 0.19b 0.21a 0.005 <0.001 <0.001 0.54
6-wk-old weight uniformity 71.40% 78.60% 86.20% 92.60% - - - -
Uniformity of tibial length at 6 wk 100.00% 100.00% 100.00% 96.30% - - - -
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Abbreviations: ADFI, average daily feed intake; ADG, average daily gain; F/G, feed/body mass ratio; FBW, final body weight; IBW, initial body weight; Met, methionine; SAA, sulfur-containing amino acid; SEM, standard error of the means.

a–d

Means in the same row with different letters differ significantly, P < 0.05.

0.43% Met as the control group.

Effect of Dietary Met Content on the Serum Proteins of Hyline Grey Chicks

The diets had significant effects on the serum ALB concentration and ALP activity at 14 days old. The inclusion of Met at 0.43% or 0.54% in the diet increased the ALB concentration (P < 0.001), and 0.54% Met significantly increased A/G compared versus the control group (0.31% Met) (P < 0.05). However, the feeding of 0.54% Met resulted in significantly lower ALP activity than 0.31% Met (Figure 1).

Figure 1.

Figure 1

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Effect of Met levels on protein metabolism indicators in the serum of chicks at 14 d of age. Concentration of these indices TP (A), ALB (B), GLB (C), ALB/GLB (D), LD (E), UA (F), UREA (G), GLU (H), and ALP (I) in the serum at 14 d of age. 0.43% Met as the control group. Data were presented as means ± SEM (n = 6 per group). Statistical significance was determined using one-way ANOVA, followed by Turkey test. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001.

High dietary Met levels (0.43% Met and 0.54% Met) also significantly affected the serum protein concentrations of ALB and A/G, and the ALP activity, at 42 days old (Figure 4). In addition, high Met levels in the diet significantly increased the serum ALB concentration and A/G ratio (P < 0.05) and significantly reduced the serum ALP activity (P < 0.001).

Figure 4.

Figure 4

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Effect of Met levels on protein metabolism indicators in the serum of chicks at 42 d of age. Concentration of these indices TP (A), ALB (B), GLB (C), ALB/GLB (D), LD (E), UA (F), UREA (G), GLU (H), and ALP (I) in the serum at 42 d of age. 0.43% Met as the control group. Data were presented as means ± SEM (n = 6 per group). Statistical significance was determined using one-way ANOVA, followed by Turkey test. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001.

Effect of Met on Serum Indices of Immunity in Hyline Grey Chicks

The Met content of the diet had significant effects on the serum concentrations of TNF-α and IgM at 14 days old. High levels of Met (0.43% and 0.54%) in the diet significantly increased the IgM concentration (P < 0.05). Moreover, the TNF-α concentration of the 0.38% Met group was significantly lower than that of the control (0.31% Met) and 0.43% Met groups (P < 0.05). The highest IL-1β concentrations were measured in the 0.38% Met group, and the lowest concentrations of the pro-inflammatory mediators IL-1β and IL-6 were present in the 0.54% Met group. Furthermore, the highest serum concentrations of the anti-inflammatory cytokine IL-10 were present in the 0.54% Met group (Figure 2).

Figure 2.

Figure 2

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Effect of Met levels on immunity related indicators in the serum of chicks at 14 d of age. Concentration of these indices IL-1β (A), IL-6 (B), TNF-α (C), IL-10 (D), IgA (E), IgM (F), and IgG (G) in the serum at 14 d of age. 0.43% Met as the control group. Data were presented as means ± SEM (n = 6 per group). Statistical significance was determined using one-way ANOVA, followed by Turkey test. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001.

The dietary Met content also had significant effects (Figure 5) on the serum concentrations of IL-1β, IL-6, TNF-α, IL-10, IgA, and IgG in the 42-day-old layers. The feeding of diets containing 0.43% or 0.54% Met was associated with significantly lower IL-1β, IL-6, and TNF-α concentrations (P < 0.05), but a significantly higher concentration of the anti-inflammatory cytokine IL-10 (P < 0.05).

Figure 5.

Figure 5

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Effect of Met levels on immunity-related indicators in the serum of chicks at 42 d of age. Concentration of these indices IL-1β (A), IL-6 (B), TNF-α (C), IL-10 (D), IgA (E), IgM (F), and IgG (G) in the serum at 42 d of age. 0.43% Met as the control group. Data were presented as means ± SEM (n = 6 per group). Statistical significance was determined using one-way ANOVA, followed by Turkey test. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001.

Effects of Met on Serum Indices of Oxidative Stress in Hyline Grey Chicks

Significant effects of Met on the serum indices of oxidative status, including T-SOD, T-GSH, H2O2, and ·OH, were observed in 14-day-old chicks. Met at 0.43% or 0.54% in the diet significantly increased the activity of the antioxidants T-SOD and T-GSH (P < 0.05) and improved the antioxidant status of the chicks by significantly reducing the H2O2 and ·OH concentrations (P < 0.05) (Figure 3). The dietary Met content also had a significant effect (Figure 6) on the serum concentrations of T-SOD, MDA, GSH-PX, T-AOC, T-GSH, H2O2, and ·OH in 42-day-old chicks. Diets containing 0.43% or 0.54% Met significantly increased the activities of T-SOD, GSH-PX, T-AOC, and T-GSH (P < 0.05), while significantly reducing the serum ·OH concentration (P < 0.05). Moreover, the serum MDA concentration of the 0.43% Met group was significantly lower than that of the 0.38% Met group (P < 0.05). Finally, the H2O2 concentration was significantly lower in the 0.54% Met group then in the control group (0.31% Met) (P < 0.05).

Figure 3.

Figure 3

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Effect of Met levels on oxidative stress indicators in the serum of chicks at 14 d of age. Concentration of these indices T-SOD (A), MDA (B), GSH-PX (C), T-AOC (D), CAT (E), T-GSH (F), H2O2 (G), and .OH (H) in the serum at 14 d of age. 0.43% Met as the control group. Data were presented as means ± SEM (n = 6 per group). Statistical significance was determined using one-way ANOVA, followed by Turkey test. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001.

Figure 6.

Figure 6

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Effect of Met levels on oxidative stress indicators in the serum of chicks at 42 d of age. Concentration of these indices IL-1β (A), IL-6 (B), TNF-α (C), IL-10 (D), IgA (E), IgM (F), and IgG (G) in the serum at 42 d of age. 0.43% Met as the control group. Data were presented as means ± SEM (n = 6 per group). Statistical significance was determined using one-way ANOVA, followed by Turkey test. * P ≤ 0.05, ** P ≤ 0.01, *** P ≤ 0.001.

Effect of Met on the Growth Performance of Hyline Grey Hens

As the dietary Met content was increased during the brooding period, there were linear increases in BM and TL at wk 8 (P < 0.05). At 15 wk old, the TL was significantly higher in the 0.38% Met group than in the other 3 treatment groups (P < 0.05); and at 20 wk old, the TL was significantly higher in the 0.43% Met group than in the other 3 groups (P < 0.05) (Figure 7).

Figure 7.

Figure 7

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Effect of Met levels on subsequent growth performance of laying hens. Means with different superscripts within a column differ significantly (P < 0.05). SEM: standard error of the means. Met, methionine. 0.43% Met as the control group.

DISCUSSION

In the present study, the effects of various levels of dietary Met content during the brooding period were compared, focusing on the growth and development, protein metabolism, immune status, oxidative stress status, and subsequent growth performance of chicks. Met is the main limiting amino acid in corn-soya meal-based laying hen diets (Rama Rao et al., 2019; Beheshti Moghadam et al., 2021), and provides organic sulfur elements. Met has been shown to be indispensable for protein synthesis, the growth and development, and the immune function of animals, and either an excess or a deficiency of Met affects the growth performance of poultry (Elango, 2020a; Savino et al., 2022). In the present study, Met supplementation improved the growth performance, immune status, and antioxidant capacity of chicks. Furthermore, the study revealed that the feeding of diets containing relatively high levels of Met during the early stages of growth have sustained effects on the growth of poultry. These findings provide a theoretical basis for and a new perspective on the feeding of chicks to ensure their optimal growth performance as laying hens, which should improve the economic efficiency of egg production.

It is well known that laying hens during the brooding period have greater nutritional requirements than growing layers (Macelline et al., 2021). Thus, chickens require adequate nutritional support during the brooding period to ensure optimal growth performance. The growth performance of laying hens is assessed using a range of indices, such as BM, ADG, ADFI, and F/G. Supplementation of poultry diets with Met significantly increases feed intake, promotes the growth and development of laying hens, and leads to better feed conversion rates. Conversely, a deficiency in dietary Met causes a marked reduction in feed intake, and therefore relatively poor growth performance (Castro et al., 2020). The study revealed that the addition of Met to the diet for 6 wk has positive effects on the ADG, F/G, and FBW of layers during the brooding period, but no effects on IBM or ADFI. The ADG and FBW of the 0.43% and 0.54% Met groups were significantly higher than those of the control group (0.31% Met). The group receiving 0.54% Met exhibited a lower F/G ratio and more uniform body weights at 6 wk of age compared to the control group. These results imply that the level of Met supplementation used in the study, although higher than the NRC-recommended values, does not have a negative effect on the growth performance of female chicks, but instead promotes their growth and development during the brooding period. However, the data did not show a significant decreasing trend, and therefore additional trials are needed to investigate this issue further.

A number of serum biochemical parameters reflect the metabolism, nutritional status, and the physiological and pathological changes taking place in the body. Dietary amino acid deficiencies negatively affect egg production, egg quality, and serum biochemical indices (Geng et al., 2021), and a deficiency of Met affects protein synthesis (Elango, 2020a; Liu et al., 2022; Teng et al., 2023). When animals are healthy, protein synthesis increases, and the serum TP and ALB concentrations also tend to increase (Ma et al., 2021). UA is an end product of catabolism (Chen et al., 2018), and the serum UA concentration reflects the health status of the animal. Ma et al. showed that high dietary Met levels (0.31–0.47%) are associated with greater amino acid utilization and alterations in the serum UA concentration (Ma et al., 2021). As the dietary level of methionine (Met) increased, corresponding increases in the serum concentrations/activities of TP, ALB, and ALP were observed. This is consistent with the above findings and implies that amino acid intake influences protein synthesis. However, no clear trends were observed in UA concentration, potentially due to the interaction of higher levels of Met with other amino acids or their influence on the pathways of amino acid utilization. Because the interactions between the various amino acids have not been thoroughly studied, further research is needed to better understand these.

A lack of Met in the diet affects immunity (Wu et al., 2013; Bian et al., 2020). For example, Ma et al. showed that hens fed a relatively large amount of Met were metabolically active and showed superior immune responses (Ma et al., 2021). Immunoglobulins play an important role in humoral immunity (Ulmer-Franco et al., 2012; Megha and Mohanan, 2021), and in the present study, Met supplementation increased the serum concentrations of the anti-inflammatory cytokine IL-10, as well as those of IgA and IgM in 14-day-old chicks, which suggests that Met may have a positive effect on their immune status. Interestingly, at 42 days old, the serum concentration of IL-10 was still higher in birds fed a high-Met diet, and the IgA and IgM concentrations negatively correlated with dietary Met content, implying that improves immune function. In contrast, the serum concentrations of the pro-inflammatory cytokines IL-1β and IL-6 negatively correlated with the Met level in the diet. However, dietary Met did not affect the serum concentrations of urea, UA, TNF-α, or IgG, which the speculation is that this may be related to the level of metabolism of these substances.

The imbalance between the production and elimination of reactive oxygen species (ROS) is an important cause of oxidative stress (Sies and Jones, 2020; Wang et al., 2020; Xu et al., 2022). Reactive oxygen species can cause damage to cellular components, such as DNA, proteins, and carbohydrates, resulting in tissue damage (Halliwell, 2007; Sies and Jones, 2020). Previous studies have shown that supplementation of the diet with specific amounts of DL-methionine increases the SOD and T-AOC activities and reduces the glutathione concentration (Reda et al., 2020). Met is an essential component of the cellular antioxidant system and is vital for the maintenance of normal cell function and health (Colovic et al., 2018). In the present study, Met increased the activities/concentrations of T-SOD, GSH-PX, T-AOC, and T-GSH, indicating that it improves the antioxidant capacity of the birds, consistent with the results of previous studies. Many previous studies have shown that hydrogen peroxide acts as an intracellular and intercellular signaling molecule and affects the entire process from embryonic development to cell death (Stone and Yang, 2006; Palomero and Jackson, 2010; Rice, 2011). The study revealed that the serum H2O2 concentration of the chicks tended to decrease with increasing Met concentration at both 14 and 42 days old, suggesting that increasing the dietary Met concentration might affect intracellular and intercellular signaling in the organism. Hydroxyl radicals, which are potent oxidizing agents, can kill red blood cells and cause the degradation of DNA, cell membranes, and polysaccharide compounds (Dizdaroglu and Jaruga, 2012; Andrés et al., 2023). The study revealed that increasing dietary methionine (Met) levels significantly reduced serum hydroxyl radical (·OH) concentrations in chicks at both 14 and 42 d of age, indicating potential health benefits. It has been reported that chronic inflammatory diseases involve biomolecular damage caused by ROS, including ·OH (Komaki et al., 2019), and therefore this effect of Met supplementation would be expected to reduce inflammation and biomolecular damage, and thereby facilitate healthy growth and development.

The brooding period is crucial for the subsequent growth and development of layers (Koelkebeck et al., 1987). As an illustration, dietary supplementation with an appropriate level of Met significantly improves growth performance, amino acid metabolism, and intestinal development, as well as later egg production (Liu et al., 2022). In the present study, linear increases in BM and TL were found as the dietary level of Met was increased in 8-wk-old chicks. In addition, at 15 wk old, the TL was significantly higher in the 0.38% Met group than in the other groups, whereas at 20 wk, the TL was significantly higher in the 0.43% Met group. In addition, the dietary Met level influenced subsequent body weight uniformity to some extent. Interestingly, the 0.54% Met group had more uniform body weights at 8 to 15 wk, while at 20 wk old, the 0.38% Met group had slightly higher uniformity. This suggests that feeding 0.43 to 0.54% Met-containing diets from early in life influences the subsequent growth and development of the hens, possibly because the early supplementation of Met facilitates sufficient nutrient accumulation, but the mechanism involved requires further investigation.

In conclusion, it has been demonstrated that the inclusion of 0.43 to 0.54% Met in the diet of Hyline Grey chicks 0.43 to 0.54% Met during the brooding period significantly improved their growth performance, indicated by higher ADG and FBW and lower F/G. Interestingly, the results from the present study also demonstrated that dietary 0.43 to 0.54% Met level had beneficial effects on metabolism and antioxidant systems. However, future studies should aim to identify the mechanisms whereby Met affects the physiology of these chicks during the brooding period.

ACKNOWLEDGMENTS

Acknowledgment is given to all technicians at the experimental animal facility of China Agricultural University for their diligent daily care of the laying hens involved in this study.

This research was funded by the National Key Research and Development Programs of China (2022YFD1300503) and a Special Fund for China Agricultural Research System program (grant no. CARS-40-K08).

Author Contributions: The authors’ responsibilities were as follows: JTZ, SMH, QGM and SJG designed the study. JTZ, SJG, YHZ and SMH conducted the experiments and draft the manuscript. JTZ, SJG, YHZ, LL, QXL, YFL, and SMH polished the manuscript and finished the submission. JTZ, SJG, LL, SMH, YHZ and QGM guided to analyze the experimental data. SMH, LHZ, and QGM helped with revisiting and reviewing the manuscript. All authors read and approved the final manuscript.

Consent for Publication: All authors of this work concur with this submission, and the data presented have not been previously reported nor are they under consideration for publication elsewhere.

DISCLOSURES

The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

References

  1. Abdel-Moneim A.E., Shehata A.M., Paswan V.K. Editorial: Early life programming in poultry: recent insights and interventional approaches. Front. Vet. Sci. 2022;9 doi: 10.3389/fvets.2022.1105653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Andrés C.M.C., Lastra J.M.P., Juan C.A., Plou F.J., Pérez-Lebeña E. Chemical insights into oxidative and nitrative modifications of DNA. Int. J. Mol. Sci. 2023;24:15240. doi: 10.3390/ijms242015240. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barekatain R., Kluenemann M. Intestinal permeability, microbiota composition, and expression of genes related to intestinal barrier function of broiler chickens fed different methionine sources supplemented at varying concentrations. Poult. Sci. 2023;102 doi: 10.1016/j.psj.2023.102656. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Beheshti Moghadam M.H., Aziza A.E., Cherian G. Choline and methionine supplementation in layer hens fed flaxseed: effects on hen production performance, egg fatty acid composition, tocopherol content, and oxidative stability. Poult. Sci. 2021;100 doi: 10.1016/j.psj.2021.101299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bian Y., Li W., Kremer D.M., Sajjakulnukit P., Li S., Crespo J., Nwosu Z.C., Zhang L., Czerwonka A., Pawłowska A., Xia H., Li J., Liao P., Yu J., Vatan L., Szeliga W., Wei S., Grove S., Liu J.R., Mclean K., Cieslik M., Chinnaiyan A.M., Zgodziński W., Wallner G., Wertel I., Okła K., Kryczek I., Lyssiotis C.A., Zou W. Cancer slc43a2 alters t cell methionine metabolism and histone methylation. Nature. 2020;585:277–282. doi: 10.1038/s41586-020-2682-1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brosnan J.T., Brosnan M.E. The sulfur-containing amino acids: an overview. J. Nutr. 2006;136:1636s–1640s. doi: 10.1093/jn/136.6.1636S. [DOI] [PubMed] [Google Scholar]
  7. Castro F.L.S., Kim Y., Xu H., Kim W.K. The effect of total sulfur amino acid levels on growth performance and bone metabolism in pullets under heat stress. Poult. Sci. 2020;99:5783–5791. doi: 10.1016/j.psj.2020.06.081. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chen N.N., Liu B., Xiong P.W., Guo Y., He J.N., Hou C.C., Ma L.X., Yu D.Y. Safety evaluation of zinc methionine in laying hens: Effects on laying performance, clinical blood parameters, organ development, and histopathology. Poult. Sci. 2018;97:1120–1126. doi: 10.3382/ps/pex400. [DOI] [PubMed] [Google Scholar]
  9. Colovic M.B., Vasic V.M., Djuric D.M., Krstic D.Z. Sulphur-containing amino acids: protective role against free radicals and heavy metals. Curr. Med. Chem. 2018;25:324–335. doi: 10.2174/0929867324666170609075434. [DOI] [PubMed] [Google Scholar]
  10. Dizdaroglu M., Jaruga P. Mechanisms of free radical-induced damage to DNA. Free Radic. Res. 2012;46:382–419. doi: 10.3109/10715762.2011.653969. [DOI] [PubMed] [Google Scholar]
  11. Elango R. Methionine nutrition and metabolism: insights from animal studies to inform human nutrition. J. Nutr. 2020;150:2518s–2523s. doi: 10.1093/jn/nxaa155. [DOI] [PubMed] [Google Scholar]
  12. Elango R. Methionine nutrition and metabolism: insights from animal studies to inform human nutrition. J. Nutr. 2020;150:2518S–2523S. doi: 10.1093/jn/nxaa155. [DOI] [PubMed] [Google Scholar]
  13. Fagundes N.S., Milfort M.C., Williams S.M., Da Costa M.J., Fuller A.L., Menten J.F., Rekaya R., Aggrey S.E. Dietary methionine level alters growth, digestibility, and gene expression of amino acid transporters in meat-type chickens. Poult. Sci. 2020;99:67–75. doi: 10.3382/ps/pez588. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Geng S., Huang S., Ma Q., Li F., Gao Y., Zhao L., Zhang J. Alterations and correlations of the gut microbiome, performance, egg quality, and serum biochemical indexes in laying hens with low-protein amino acid-deficient diets. ACS Omega. 2021;6:13094–13104. doi: 10.1021/acsomega.1c00739. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Grimble R.F. The effects of sulfur amino acid intake on immune function in humans. J. Nutr. 2006;136:1660s–1665s. doi: 10.1093/jn/136.6.1660S. [DOI] [PubMed] [Google Scholar]
  16. Haiden N., Haschke F. Early nutrition must be safe and should have positive impacts on long-term health. Nutrients. 2023;15:2645. doi: 10.3390/nu15122645. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Halliwell B. Biochemistry of oxidative stress. Biochem. Soc. Trans. 2007;35:1147–1150. doi: 10.1042/BST0351147. [DOI] [PubMed] [Google Scholar]
  18. Jin S., He L., Yang C., He X., Chen H., Feng Y., Tang W., Li J., Liu D., Li T. Crosstalk between trace elements and t-cell immunity during early-life health in pigs. Sci. China Life Sci. 2023;66:1994–2005. doi: 10.1007/s11427-022-2339-0. [DOI] [PubMed] [Google Scholar]
  19. Koelkebeck K.W., Amoss M.S., Jr., Cain J.R. Production, physiological, and behavioral responses of laying hens in different management environments. Poult. Sci. 1987;66:397–407. doi: 10.3382/ps.0660397. [DOI] [PubMed] [Google Scholar]
  20. Komaki Y., Simpson A.M., Choe J.K., Pinney M.M., Herschlag D., Chuang Y.H., Mitch W.A. Serum electrolytes can promote hydroxyl radical-initiated biomolecular damage from inflammation. Free Radic. Biol. Med. 2019;141:475–482. doi: 10.1016/j.freeradbiomed.2019.07.023. [DOI] [PubMed] [Google Scholar]
  21. Li P., He W., Wu G. Composition of amino acids in foodstuffs for humans and animals. Adv. Exp. Med. Biol. 2021;1332:189–210. doi: 10.1007/978-3-030-74180-8_11. [DOI] [PubMed] [Google Scholar]
  22. Li Q., Chen J., Liu J., Lin T., Liu X., Zhang S., Yue X., Zhang X., Zeng X., Ren M., Guan W., Zhang S. Leucine and arginine enhance milk fat and milk protein synthesis via the casr/g(i)/mtorc1 and casr/g(q)/mtorc1 pathways. Eur. J. Nutr. 2023;62:2873–2890. doi: 10.1007/s00394-023-03197-7. [DOI] [PubMed] [Google Scholar]
  23. Lilburn M.S., Loeffler S. Early intestinal growth and development in poultry. Poult. Sci. 2015;94:1569–1576. doi: 10.3382/ps/pev104. [DOI] [PubMed] [Google Scholar]
  24. Liu Y., Wang D., Zhao L., Zhang J., Huang S., Ma Q. Effect of methionine deficiency on the growth performance, serum amino acids concentrations, gut microbiota and subsequent laying performance of layer chicks. Front. Vet. Sci. 2022;9:878107. doi: 10.3389/fvets.2022.878107. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Ma M., Geng S., Liu M., Zhao L., Zhang J., Huang S., Ma Q. Effects of different methionine levels in low protein diets on production performance, reproductive system, metabolism, and gut microbiota in laying hens. Front. Nutr. 2021;8 doi: 10.3389/fnut.2021.739676. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Macelline S.P., Toghyani M., Chrystal P.V., Selle P.H., Liu S.Y. Amino acid requirements for laying hens: a comprehensive review. Poult. Sci. 2021;100 doi: 10.1016/j.psj.2021.101036. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Majdeddin M., Golian A., Kermanshahi H., Michiels J., De Smet S. Effects of methionine and guanidinoacetic acid supplementation on performance and energy metabolites in breast muscle of male broiler chickens fed corn-soybean diets. Br. Poult. Sci. 2019;60:554–563. doi: 10.1080/00071668.2019.1631447. [DOI] [PubMed] [Google Scholar]
  28. Martínez Y., Li X., Liu G., Bin P., Yan W., Más D., Valdivié M., Hu C.A., Ren W., Yin Y. The role of methionine on metabolism, oxidative stress, and diseases. Amino Acids. 2017;49:2091–2098. doi: 10.1007/s00726-017-2494-2. [DOI] [PubMed] [Google Scholar]
  29. Megha K.B., Mohanan P.V. Role of immunoglobulin and antibodies in disease management. Int. J. Biol. Macromol. 2021;169:28–38. doi: 10.1016/j.ijbiomac.2020.12.073. [DOI] [PubMed] [Google Scholar]
  30. Millecam J., Khan D.R., Dedeurwaerder A., Saremi B. Optimal methionine plus cystine requirements in diets supplemented with l-methionine in starter, grower, and finisher broilers. Poult. Sci. 2021;100:910–917. doi: 10.1016/j.psj.2020.11.023. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Palomero J., Jackson M.J. Redox regulation in skeletal muscle during contractile activity and aging. J. Anim. Sci. 2010;88:1307–1313. doi: 10.2527/jas.2009-2436. [DOI] [PubMed] [Google Scholar]
  32. Rama Rao S., Paul S.S., Raju M., Prakash B., Srilatha T. Effect of incremental levels of dietary total sulfur amino acids on performance of white leghorn laying hens fed diets containing guar (cyamopsis tetragonoloba) meal. J. Sci. Food Agric. 2019;99:3086–3096. doi: 10.1002/jsfa.9522. [DOI] [PubMed] [Google Scholar]
  33. Ratsika A., Codagnone M.C., O’mahony S., Stanton C., Cryan J.F. Priming for life: early life nutrition and the microbiota-gut-brain axis. Nutrients. 2021;13:423. doi: 10.3390/nu13020423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Reda F.M., Swelum A.A., Hussein E.O.S., Elnesr S.S., Alhimaidi A.R., Alagawany M. Effects of varying dietary dl-methionine levels on productive and reproductive performance, egg quality, and blood biochemical parameters of quail breeders. Animals (Basel) 2020;10 doi: 10.3390/ani10101839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Rice M.E. H2O2: a dynamic neuromodulator. Neuroscientist. 2011;17:389–406. doi: 10.1177/1073858411404531. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Savino R.J., Kempisty B., Mozdziak P. The potential of a protein model synthesized absent of methionine. Molecules. 2022;27:3679. doi: 10.3390/molecules27123679. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Sekiz S.S., Scott M.L., Nesheim M.C. The effect of methionine deficiency on body weight, food and energy utilization in the chick. Poult. Sci. 1975;54:1184–1188. doi: 10.3382/ps.0541184. [DOI] [PubMed] [Google Scholar]
  38. Sies H., Jones D.P. Reactive oxygen species (ros) as pleiotropic physiological signalling agents. Nat. Rev. Mol. Cell Biol. 2020;21:363–383. doi: 10.1038/s41580-020-0230-3. [DOI] [PubMed] [Google Scholar]
  39. Stipanuk M.H. Metabolism of sulfur-containing amino acids: how the body copes with excess methionine, cysteine, and sulfide. J. Nutr. 2020;150:2494s–2505s. doi: 10.1093/jn/nxaa094. [DOI] [PubMed] [Google Scholar]
  40. Stone J.R., Yang S. Hydrogen peroxide: a signaling messenger. Antioxid. Redox Signal. 2006;8:243–270. doi: 10.1089/ars.2006.8.243. [DOI] [PubMed] [Google Scholar]
  41. Taha-Abdelaziz K., Hodgins D.C., Lammers A., Alkie T.N., Sharif S. Effects of early feeding and dietary interventions on development of lymphoid organs and immune competence in neonatal chickens: a review. Vet. Immunol. Immunopathol. 2018;201:1–11. doi: 10.1016/j.vetimm.2018.05.001. [DOI] [PubMed] [Google Scholar]
  42. Teng P.Y., Liu G., Choi J., Yadav S., Wei F., Kim W.K. Effects of levels of methionine supplementations in forms of l- or dl-methionine on the performance, intestinal development, immune response, and antioxidant system in broilers challenged with Eimeria spp. Poult. Sci. 2023;102 doi: 10.1016/j.psj.2023.102586. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Ullrich C., Langeheine M., Brehm R., Taube V., Rosillo Galera M., Rohn K., Popp J., Visscher C. Influence of different methionine sources on performance and slaughter characteristics of broilers. Animals (Basel) 2019;9:984. doi: 10.3390/ani9110984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Ulmer-Franco A.M., Cherian G., Quezada N., Fasenko G.M., Mcmullen L.M. Hatching egg and newly hatched chick yolk sac total igy content at 3 broiler breeder flock ages. Poult. Sci. 2012;91:758–764. doi: 10.3382/ps.2011-01757. [DOI] [PubMed] [Google Scholar]
  45. Wang Y., Chen Y., Zhang X., Lu Y., Chen H. New insights in intestinal oxidative stress damage and the health intervention effects of nutrients: A review. J. Funct. Foods. 2020;75 [Google Scholar]
  46. Wang S.-H., Wang W.-W., Zhang H.-J., Wang J., Chen Y., Wu S.-G., Qi G.-H. Conjugated linoleic acid regulates lipid metabolism through the expression of selected hepatic genes in laying hens. Poult. Sci. 2019;98:4632–4639. doi: 10.3382/ps/pez161. [DOI] [PubMed] [Google Scholar]
  47. Wen C., Jiang X.Y., Ding L.R., Wang T., Zhou Y.M. Effects of dietary methionine on growth performance, meat quality and oxidative status of breast muscle in fast- and slow-growing broilers. Poult. Sci. 2017;96:1707–1714. doi: 10.3382/ps/pew432. [DOI] [PubMed] [Google Scholar]
  48. Wu B., Cui H., Peng X., Fang J., Cui W., Liu X. Pathology of bursae of fabricius in methionine-deficient broiler chickens. Nutrients. 2013;5:877–886. doi: 10.3390/nu5030877. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Wu G. Amino acids: metabolism, functions, and nutrition. Amino Acids. 2009;37:1–17. doi: 10.1007/s00726-009-0269-0. [DOI] [PubMed] [Google Scholar]
  50. Wu G. Functional amino acids in growth, reproduction, and health. Adv. Nutr. 2010;1:31–37. doi: 10.3945/an.110.1008. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Wu G., Bazer F.W., Dai Z., Li D., Wang J., Wu Z. Amino acid nutrition in animals: protein synthesis and beyond. Annu. Rev. Anim. Biosci. 2014;2:387–417. doi: 10.1146/annurev-animal-022513-114113. [DOI] [PubMed] [Google Scholar]
  52. Xu X., Wei Y., Hua H., Jing X., Zhu H., Xiao K., Zhao J., Liu Y. Polyphenols sourced from ilex latifolia thunb. Relieve intestinal injury via modulating ferroptosis in weanling piglets under oxidative stress. Antioxidants (Basel) 2022;11:966. doi: 10.3390/antiox11050966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Zeitz J.O., Fleischmann A., Ehbrecht T., Most E., Friedrichs S., Whelan R., Gessner D.K., Failing K., Lütjohann D., Eder K. Effects of supplementation of dl-methionine on tissue and plasma antioxidant status during heat-induced oxidative stress in broilers. Poult. Sci. 2020;99:6837–6847. doi: 10.1016/j.psj.2020.08.082. [DOI] [PMC free article] [PubMed] [Google Scholar]

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