World's Poultry Science Journal

ISSN: (Print) (Online) Journal homepage: https://www.tandfonline.com/loi/twps20

Physiological role of Arginine in growth

performance, gut health and immune response in
broilers: a review

F. Hassan, M. A. Arshad, S. Hassan, R. M. Bilal, M. Saeed & M. S. Rehman

To cite this article: F. Hassan, M. A. Arshad, S. Hassan, R. M. Bilal, M. Saeed & M. S.

Rehman (2021) Physiological role of Arginine in growth performance, gut health and
immune response in broilers: a review, World's Poultry Science Journal, 77:3, 517-537, DOI:
10.1080/00439339.2021.1925198

To link to this article: https://doi.org/10.1080/00439339.2021.1925198

Published online: 24 May 2021.

Submit your article to this journal

Article views: 565

View related articles

View Crossmark data

Citing articles: 3 View citing articles

Full Terms & Conditions of access and use can be found at

https://www.tandfonline.com/action/journalInformation?journalCode=twps20
WORLD’S POULTRY SCIENCE JOURNAL
2021, VOL. 77, NO. 3, 517–537
https://doi.org/10.1080/00439339.2021.1925198

Physiological role of Arginine in growth performance, gut

health and immune response in broilers: a review
a
F. Hassan *, M. A. Arshada*, S. Hassana, R. M. Bilalb, M. Saeed c
and M. S. Rehmana
a
Institute of Animal and Dairy Sciences, Faculty of Animal Husbandry, University of Agriculture, Faisalabad,
Pakistan; bCollege of Veterinary and Animal Sciences, The Islamia University of Bahawalpur, Bahawalpur,
Pakistan; cFaculty of Animal Production and Technology, Cholistan University of Veterinary and Animal
Sciences, Bahawalpur, Pakistan

SUMMARY KEYWORDS
Arginine (Arg) is one of the metabolically versatile essential amino Arginine; feed additive;
acids. Dietary supplementation with Arg has been shown to stimu­ growth; gut health;
late the secretion of insulin-like growth factor while improving immunity; super dosing
growth performance and feed efficiency in broilers. In addition to
its role in protein synthesis, Arg is also a precursor of nitric oxide,
creatine, and polyamines. It also modulates lipid metabolism by
reducing total body fat accumulation to improve meat quality and
antioxidant defence. Moreover, it is considered as an essential amino
acid for chicks due to the absence of a functional urea cycle in birds
at an early age. Arginine plays a crucial role in metabolic pathways
associated with immune-competence and growth. Dietary supple­
mentation of Arg at 12 to 15 g/kg of diet resulted in linear increase in
body weight gain from 7.5 to 17% in broilers fed soybean meal-
based diets. Arg acts as a key vasodilator that opposes the onset of
pulmonary hypertension in broilers and is particularly beneficial
under high altitude and hypoxia conditions. Dietary Arg supplemen­
tation reduces ascites-related mortality under low ambient tempera­
tures and attenuates adverse effects of heat stress and high stock
density. Moreover, in ovo feeding of Arg increased levels of secretory
immunoglobulin A revealing its potential to modulate immune bar­
rier function leading to enhanced overall immunity and intestinal
health of birds. The NRC recommended levels of Arg are 1.25%,
1.10%, and 1.00% (of the broiler diet from 1–3, 4–6 and 7–8 weeks,
respectively). However, other studies have reported 101, 103 and
107% of NRC recommendation of Arg for maximum feed efficiency,
growth performance, and optimal immune function, from 1–3, 4–6
and 7–8 weeks, respectively, under thermoneutral conditions. This
review provides insights into the optimal supplementation of Arg
above NRC recommendations to improve growth performance, meat
quality, and immunity of broilers.

Introduction
Amino acids are the building blocks of protein and are essential for development of body
tissues and immune organs. Deficiency of amino acids can lead to reduced growth

CONTACT M. S. Rehman shsaifurrehman@yahoo.com Institute of Animal and Dairy Sciences, Faculty of Animal
Husbandry, University of Agriculture, Faisalabad 38040, Pakistan
*These authors contributed equally to this work
© 2021 World’s Poultry Science Association.
518 F. HASSAN ET AL.

performance and stress on the immune system (Rubin et al. 2007; Birmani et al. 2019).
Therefore, optimal nutritional strategies are recommended to positively affect the
immune system. It is well established that dietary protein and amino acid levels can
modulate the immune system (Abdukalykova and Ruiz-Feria 2006; Ruiz-Feria 2009).
Adequate quantity and quality of dietary protein is required for growth performance and
proper cell-mediated immunity, as well as secretory immunoglobulin, antibody and
cytokine production (Li et al. 2007; Birmani et al. 2019). However, protein deficiency
ultimately reduces the antibody production and the development of antibody-producing
cells in response to T-dependent antigens (Woodward 1998).
Synthetic amino acids are used in broiler diets to obtain optimum performance.
Arginine is an essential amino acid that is required in many important biological and
physiological functions in the body (Ball, Urschel, and Pencharz 2007; Castro and Kim
2020). It is vital for optimal growth performance and supplies creatine in the body
(Fouad et al. 2012; Khajali and Wideman 2010). Arginine also improves digestive
functions in birds and mammals by reducing intestinal permeability due to its ability
to synthesise nitric oxide (NO) (Zhang et al. 2017; Castro et al. 2020). Moreover, it is
involved in the healing of gastrointestinal ulceration (Wideman et al. 2007; Bortoluzzi,
Rochell, and Applegate 2017). Many adult mammals can synthesise Arg according to
their body requirements, but chickens cannot synthesise Arg due to the absence of
carbamoyl phosphate synthase-I (a key enzyme involved in the synthesis of Arg) and
lower activities of ornithine transcarbamoylase and hepatic arginase (Khajali, Faraji, and
Basoo 2013). Therefore, the chicken body requires an adequate amount of Arg from
dietary sources. Hence, a sufficient amount of Arg must be present in the feed of chickens
to maintain protein accretion, immunological and physiological functions (Khajali and
Wideman 2010). Dietary Arg also promotes growth rate in chickens (Ball, Urschel, and
Pencharz 2007), as it increases protein synthesis and secretion of hormones such as
insulin, glucagon, and growth hormone which may increase feed intake and synthesis of
ornithine which further leads to enhanced synthesis of DNA and cell proliferation
(Kwak, Austic, and Dietert 1999).
The National Research Council (NRC) recommends 12.5 and 11.0 g Arg/kg of the diet
from 1–3 and 4–6 weeks, respectively (NRC 1994). Arginine supplementation beyond the
NRC recommendation has been shown to improve growth performance in broilers
(Fernandes et al. 2009; Khajali et al. 2011). Khajali, Faraji, and Basoo (2013) showed
that growth performance was improved (P < 0.05) in broilers fed diets having 15 g Arg/kg
diet. Bulbul et al. (2013) observed higher body weight and better feed conversion ratio
(FCR) in broilers fed a diet having L- Arg at 110% of Arg requirement without affecting
the feed intake. Moreover, higher (P < 0.05) breast muscle weight and higher (P < 0.05)
proportion of thigh and heart have been reported in birds fed a diet containing 0.35% Arg
(Labadan, Hsu, and Austic 2001; Khajali et al. 2011). Extensive studies have reported
a promising effect of L- Arg on poultry production (Tables 1–Tables 3).

Arg chemistry and metabolism

Arginine possesses a 4-carbon straight-chain and distal end, capped by a complex
guanidine group (Figure 1).
Table 1. Effect of Arg supplementation on growth performance of broiler.
Level Used Results
Kidd et al. (2001) 100 and 120% of NRC Broiler diet supplemented with 0.2% Arg additional to NRC (1994) standards improved production performance under
ideal environment
Khajali et al. (2011) 0, 0.2 and 0.4% They observed higher body weight gain and better FCR for birds fed canola meal-based diet with Arg at 0.4%
Basoo et al. (2012) 0.88, 0.99, 1.1, 1.21, 1.32 and 1.43% Birds fed diet containing Arg at 1.21% resulted in higher weight gain and better FCR
Al-Daraji and Salih (2012) 0, 0.02, 0.04 and 0.06% Higher feed intake, higher body weight gain and better FCR of broiler birds by supplementation of Arg at 0.06% in their
diet
Murakami et al. (2012) 1.39, 1.49, 1.59, 1.69 and 1.79% They observed improved live weight and better FCR of birds received diet containing Arg at 1.79% without increasing the
feed intake of birds during starter phase
Mejia et al. (2012) Arg: Lys ratios (0.95 and 1.24%) Feed efficiency was improved in the birds fed diet containing Arg:Lys ratio (1.24%). However, body weight gain was
unaffected by extra Arg supplementation
Khajali, Faraji, and Basoo 10 or 15 g/Kg 15 g/Kg recommended for optimum growth performance
(2013)
Fernandes et al. (2009) 0 and 1% Arg supplementation had no effect on feed intake, weight gain and FCR in first week
Laika and Jahanian (2017) 100, 105 and 110% No effect on feed intake, weight gain but improved FCR
Nayak et al. (2016) (0.5% Arg) Weight of day-old chicks, weight gain post-hatching, BFEI and BFPR were better in Arg fed in ovo groups
Gao et al. (2017a) 0.5, 1 or 2% Hatchability was decreased but body weight was linearly increased at 21 day.
Jahanian and Khalifeh- 90, 100 or 110% of NRC level of Arg or FCR Improved by increasing Arg up to 100% of NRC recommended level, although feed intake was not influenced by
Gholi (2018) Met dietary treatments
Rochell et al. (2017) 0.74 and 1.23% Body weight gain was improved of birds received diet containing 1.23% Arg
Yu et al. (2018b) 1.19, 1.44, 1.69, 1.94 or 2.19% Linearly improved weight gain and insulin-like growth factor-I (IGF-I)
Zampiga et al. (2018) Arg:Lys as 1.15, 1.15, 1.16 and 1.171 FCR was improved in supplemented groups than control at day 12, 22, 33 and in the overall period
Xu et al. (2018) 0.45, 0.90, 1.35, and 1.80% Weight gain and FCR were improved quadratically during the entire period
Sirathonpong et al. (2019) Arg:Lys ratios (0.85 and 1.26) Weight gain, FCR and insulin growth factor-I were improved
Subramaniyan et al. 100, 1000, and 2500 µg/100 µL/egg Hatchability, body weights and survival rates were better in those chicks injected Arg at 14th day of incubation
(2019)
Liu et al. (2019) 9.8, 14.7, 19.1 or 23.4 g/kg Weight gain and feed intake were quadratically increased while linearly decreased FCR
Toghyani et al. (2019) 35 (Arg), 25 (Thr) and 35 + 25 mg/egg Better feed intake and body weight in supplemented after hatching
(Arg + Thr)
WORLD’S POULTRY SCIENCE JOURNAL
519
 520

Table 2. Effect of Arg supplementation on carcase characteristics of broiler.

F. HASSAN ET AL.

Reference Level Used Results

Khajali et al. (2011) 0, 0.2 and 0.4% Birds fed Arg supplemented diet had higher carcase, breast and thigh yield. However, liver and heart weight were not
affected by Arg supplementation
Basoo et al. (2012) 0.88, 0.99, 1.1, 1.21, 1.32 and 1.43% Birds fed diet containing Arg at 1.32% resulted in higher breast yield
Mejia et al. (2012) Arg to Lys ratios (0.95 and 1.24%) Carcase, breast yield and abdominal fat were not affected by different Arg to Lys ratios
Bulbul et al., (2013) 90, 100, 110, 120 and 130% of NRC Relative weight of spleen and bursa were higher in birds fed diet containing 120 and 130% of Arg compared to control
group during starter phase
Fernandes et al. 0 and 1% Birds received diet containing Arg @ of 1% had higher intestine weight, crypt depth than control group
(2009)
Yang et al. (2016) 0, 8.5 and 17% Birds fed diet having Arg at 17 mg/kg had higher (p < 0.05) proventriculus and duodenum weight as compared to control
group
Laika and Jahanian 100, 105 and 110% Increased the carcase % and liver weight
(2017)
Gao et al. (2017a) 0.5, 1 and 2% Weight of organs was linearly increased at 21 day
Youvalari et al. CuSO4 (0, 125 & 250 ppm) and Arg (0, 0.1 Increased the breast and thigh meat but decreased the proportion of abdominal fat, lung weight
(2017) and 0.2%)
Alabi et al. (2018) 167 and 334 mg/L in the drinking water Congestion and inflammation of liver and kidney
Yu et al. (2018b) 1.19, 1.44, 1.69, 1.94 or 2.19% Linearly improved the liver weight
Zampiga et al. Arg:Lys as 1.15, 1.15, 1.16 and 1.171 Meat quality attributes, FPD occurrence and breast myopathies were remained unchanged
(2018)
Sirathonpong et al. Arg:Lys ratios Breast meat (1.26 level) and carcase yield (1.05 level) were improved
(2019) (0.85 and 1.26)
Table 3. Effect of Arg supplementation on immune response of broiler.
Reference Level Used Results
Munir et al. (2009) 2% above NRC Stimulated humoral and cell-mediated immunity in chickens vaccinated and challenged against hydropericardium
syndrome virus
Khajali et al. (2011) 0, 0.2 and 0.4% Arg supplemented diet had higher plasma NO concentration than control
Basoo et al. (2012) 0.88, 0.99, 1.1, 1.21, 1.32 and 1.43% Birds fed diet containing Arg at 1.21% resulted in higher NO level
Tan et al. (2014b) 0.99, 1.39, 1.76, 2.13 and 2.53% Increasing dietary Arg concentrations linearly and quadratically reduced the percentage of circulating B cells
Guo et al. (2015) 0, 4.5, 9, 13.5 and 18 g/kg Increasing the level of Arg from 0 to 18 g/kg in broiler diet had higher plasma NO level
Yang et al. (2016) 0, 8.5 and 17% Birds fed diet containing Arg at 8.5 mg/kg had lower triglyceride and cholesterol levels as compared to control.
Hu et al. (2016) 10, 15, 20 and 25 g/Kg Supplementation of Arg (>20.0 g/Kg) suppressed innate immunity and enhances the antioxidant activity in broiler
chickens
Kodambashi Emami GAA as 0.6 or 1.2 &/or Arg as 0.86 or Alleviated the effect of cold temperature by reduction in right ventricle to total ventricle ratio and blood haematocrit
et al. (2017) 1.72 g/Kg
Jahanian and 90, 100 or 110% of NRC level of Arg or Met Decrease in serum uric acid and increase in titre against IB and IBD
Khalifeh-Gholi
(2018)
Gao et al. (2017a) 0.5, 1 or 2% Ghrelin and glucagon-like peptide 2 concentration were increased
Gao et al. (2017b) 1% Serum iNOS activity, contents of IL-2, IL-4, NO and IgA as well as the weights of lymphoid organs in Arg group were
improved
Animashahun et al. 0, 167 or 333 mg/L Except mean corpuscular haemoglobin concentration all haematological parameters were improved
(2018)
Zampiga et al. (2018) Arg:Lys as 1.15, 1.15, 1.16 and 1.171 Higher Arg and leucine, but lower acetoacetate, glutamate, proline and adenosine concentration in plasma
Xu et al. (2018) 0.45, 0.90, 1.35, and 1.80% Serum insulin-like growth factors-1, insulin, IgA and IFN-γ as well as thymus weigh, Lymphocyte proliferation and
antibody titres against ND were improved
Toghyani et al. (2019) 35 (Arg), 25 (Thr) and 35 + 25 mg/egg (Arg Supplementation improved antibody titre against SRBC. No improvement against influenza and ND viruses. Alone Arg
+ Thr) increased bursa and spleen weights. Alone Thr increased serum albumin and glucose concentration
Subramaniyan et al. 100, 1000, and 2500 µg/100 µL/egg They observed better SGPT, SGOT, myoblast determination protein and IgM on 14th day of incubation
(2019)
Burin et al. (2019) MnSO4 or Mn with amino acid complex They observed better macrophage phagocytic activity but no difference in macrophage phagocytic activity
and Arg (1.12 or 1.20 dig.)
Liu et al. (2019) 9.8, 14.7, 19.1 and 23.4 g/kg Serum IgM and IgG contents were linearly decreased, and mRNA expression of bursal IgM and IgG were downregulated
WORLD’S POULTRY SCIENCE JOURNAL
521
522 F. HASSAN ET AL.

Figure 1. The molecular structure of Arg.

Absorption of dietary Arg occurs mainly through the intestinal epithelial layer either
by sodium-dependent or sodium-independent mechanisms, the latter being more effec­
tive (Rueda, Michelangeli, and Gonzalez-Mujica 2003; Brake and Balnave 1995).
Arginine metabolism is associated with the urea cycle in mammals (ureotelic animals).
Mammals use the enzymes of their urea cycle to synthesise L- Arg from ornithine,
ammonia, and the amino-nitrogen of aspartate (Figure 2). However, poultry lack the
key enzyme carbamoyl phosphate synthase-I and also have lower activities of hepatic
arginase and ornithine transcarbamoylase (Tamir and Ratner 1963a). Therefore, poultry
are unable to synthesise Arg and are highly dependent on dietary Arg. Accordingly, in the
broiler, Arg cannot be replaced by dietary ornithine whereas it can be replaced by dietary

Figure 2. Metabolism of Arg in chicken.

 WORLD’S POULTRY SCIENCE JOURNAL 523

citrulline to some extent. However, this sparing effect of citrulline over Arg was not
enough to support the need for Arg by the bird (Tamir and Ratner 1963b).

Dietary requirements of Arg for broilers

Arginine requirement is highly variable under different dietary protein sources in
broilers (Ball, Urschel, and Pencharz 2007). Before the NRC recommendation, the Arg
requirement was reported as being 1.10 − 1.28% of the diet for optimal growth and 0.96
to 1.28% of the diet for maximum feed efficiency (Cuca and Jensen 1990). The NRC
(1994) recommendation for Arg requirement is 1.25, 1.10, and 1.00% of the diet from
1–3, 4–6 and 6–8 weeks of broiler age, respectively. However, in various studies (Brake
and Balnave 1995; Brake 1998) Arg requirement was shown to increase during hot
weather. This is attributed mainly to the depression of both sodium-dependent and
sodium-independent mechanisms for uptake of Arg through the gut (Khajali and
Wideman 2010). Jahanian (2009) suggested an Arg requirement of 101, 103, and 107%
of NRC recommended values for maximum feed efficiency, growth performance, and
optimal immune functions, respectively, under thermo-neutral conditions. An increase
in the dietary supply of Arg and lysine (Lys) is beneficial for weight gain in broilers
during stressful conditions imposed by crowding (Srinongkote, Smriga, and Toride
2004). Enhanced muscle development was also observed in broilers during the starter
phase in response to Arg supplementation at levels above the NRC recommendations
(Fernandes et al. 2009). Recently, Castro et al. (2019) reported maximum weight gain,
lean deposition, and better FCR in Ross 308 broilers as a result of 99.68, 100.39, and
98.27% levels of Arg recommendation, respectively.

Dietary supplementation of Arg in broilers

Effect of Arg supplementation on growth performance
In ovo provision of an appropriate Arg solution has been shown to regulate early
nutrition supply and subsequent growth development in chickens (Gao et al. 2017b).
Moreover, in ovo feeding of Arg (1% Arg solution into the amnion) at 17.5 days of
incubation improved the development of digestive organs, duodenal morphology, gas­
trointestinal hormones, and mucosal enzyme activities of broiler embryos and hatchl­
ings. It subsequently led to better growth performance of chicks during the first-week
post-hatch (Gao et al. 2017a). Subramaniyan et al. (2019) reported that in ovo injection of
Arg (100, 1000, and 2500 µg/100 µL/egg) improved hatchability, body weights, and
survival rates in chicks injected with Arg on the 14th day of incubation as compared to
the 8th and 18th days. Moreover, in ovo supplementation of Arg and threonine
(35 + 25 mg/egg) improved feed intake and body weight in chicks after hatching as
compared with the unsupplemented control (Toghyani et al. 2019). Gao et al. (2017a)
reported a decrease in hatchability in response to in-ovo feeding of Arg (0.5, 1 or 2%) into
the incubating eggs of broiler breeders, but body weight was linearly increased at 21 days.
In ovo feeding of Arg has revealed far-reaching effects on the growth of broilers
throughout the rearing period leading to improved body weight gain, economy index,
and broiler feed price ratio (Nayak et al. 2016).
524 F. HASSAN ET AL.

Chamruspollert, Pesti, and Bakalli (2002) found a total Arg requirement for newly
hatched broiler chicks as 1.26% for body weight gain and 1.27% for feed conversion.
Extensive studies (Table 1) have been conducted regarding the supplementation of
dietary Arg in broilers to evaluate growth and immune response. In the majority of the
studies, a positive response of Arg supplementation on growth performance has been
reported, although to a variable extent which may be attributed to different diets, birds,
management, and environmental variations. Higher body weight gain and better FCR in
broiler chickens were observed in response to supplementation of Arg at 0.06% (Al-
Daraji and Salih 2012), 0.2% (Kidd et al. 2001), and 0.35% (Labadan, Hsu, and Austic
2001), above NRC values. Broilers fed Arg at 130% of the NRC recommendation showed
better growth performance (Khajali, Faraji, and Basoo 2013). When Eimeria challenged
broilers were fed diets supplemented with 100, 105, and 110% of the recommended Arg,
there was no effect on feed intake and weight gain; however, FCR was improved during
the starter and grower phases (Laika and Jahanian 2017). The maximum NRC recom­
mended level of Arg is 12.8 g/kg of diet; however recent studies have shown a quadratic
increase in weight gain and feed intake with Arg supplementation up to 14.7 g/kg. Levels
higher than 14.7 g/kg did not affect these traits (Liu et al. 2019) and a linear decrease in
FCR was observed in broilers as the concentration of Arg increased (L- Arg as 9.8, 14.7,
19.1 or 23.4 g/kg) in the basal diet.
Likewise, supplementation of Arg at 1.21% (Basoo et al. 2012) and 1.39 to 1.79%
(Murakami et al. 2012) improved growth performance without affecting the feed intake
of broilers (Table 1). Similarly, higher body weight gain and better FCR were observed
with a canola meal-based diet supplemented with 0.4% Arg (Khajali et al. 2011). When
Arg deficient diets were supplemented with 0.45, 0.90, 1.35, and 1.80% Arg, there was
a quadratic increase in weight gain and feed efficiency (Xu et al. 2018).
The effects of supplementation of Arg in combination with other essential amino acids
like Lys, methionine (Met), and threonine (Thr) have been evaluated in many studies.
Improved feed efficiency was observed in birds fed a diet containing higher Lys: Arg ratio
(110%) with no effect on body weight gain (Mejia et al. 2012). However, higher Lys: Arg
ratio resulted in higher weight gain in heat-stressed broilers without affecting FCR and
feed intake (Balnave and Brake 2001). Higher dietary Arg:Lys ratio (1.26) enhanced
secretion of insulin-like growth factor-I (IGF-I) in broilers leading to improved weight
gain and FCR (Sirathonpong et al. 2019). Jahanian and Khalifeh-Gholi (2018) reported
that, if the diet had a sufficient level of methionine, then Arg supplementation (90, 100,
and 110% of the NRC) improved the feed efficiency. Evaluation of different digestible
Arg:Lys ratios (1.15, 1.15, 1.16 and 1.17) in feed resulted in decreased FCR at day 12, 22,
33; however, weight gain was increased with Arg supplementation only at 33 days
compared to the control group (Zampiga et al. 2018). In ovo feeding of Arg has been
shown to regulate early energy metabolism in broilers after hatching as it significantly
increased glycogen and glucose levels in the liver and pectoral muscles; it also elevated the
plasma glucose and insulin levels while upregulating the expression of phosphoenolpyr­
uvate carboxykinase (PEPCK) and fructose1,6-bisphosphatase (FBP) mRNA (Yu et al.
2018a).
 WORLD’S POULTRY SCIENCE JOURNAL 525

Effect of Arg supplementation on carcase characteristics

Beneficial effects of Arg supplementation on carcase characteristics in broilers have been
reported in many studies (Table 2). Dietary supplementation of Arg (17 mg/kg) increased
the relative weights of proventriculus and duodenum in broilers (Yang et al. 2016).
Similarly, relative weights of spleen and bursa were higher in birds fed a diet containing
120 and 130% of Arg recommended by the NRC compared to the control group during the
starter phase (Bulbul et al. 2013). Arginine supplementation (at 1.32%) was shown to
increase breast yield in broilers (Basoo et al. 2012). Supplemental Arg above the recom­
mended NRC value (0, 0.2 and 0.4% L- Arg) in canola-based diets resulted in higher
carcase, breast, and thigh yields (Khajali et al. 2011). Fernandes et al. (2009) reported that
birds receiving a diet containing 1.0% of Arg had higher intestinal weight and crypt depth.
The abdominal fat: live body weight ratio in poultry may be used to monitor the total
deposition of body fat, since this trait is highly associated with live body weight and fat
accretion in avian species (Becker et al. 1979). Dietary supplementation of L-Arg mini­
mised the abdominal fat content by decreasing the synthesis of fatty acids (Figure 3).
Breast muscle yield was improved in response to higher dietary Arg: Lys ratio (1.26) in
broilers (Sirathonpong et al. 2019); however, the optimum dietary Arg: Lys ratio to
improve carcase yield in the grower phase was 1.05 in this study. Arginine supplementa­
tion has also been shown to increase liver weight linearly in laying hens, which can also
influence the overall metabolic efficiency of birds (Yu et al. 2018b). Moreover, in ovo
injection of L- Arg (0.5, 1 or 2%) in broiler breeders linearly increased the relative weight
of digestive organs at d 21 (Gao et al. 2017a).

Figure 3. Effect of L-Arg on deposition of protein and fat in the body.

526 F. HASSAN ET AL.

Boilers fed diets containing supplemental copper (Cu) and Arg (0.2%) showed pro­
portionally higher breast and thigh meat but lower relative abdominal fat and lung weight
(Youvalari et al. 2017). Despite the desirable effects on organ weight, Arg protected
against a decrease in the relative weight of liver and carcase percentage in Eimeria
challenged broilers (Laika and Jahanian 2017). These findings may partly explain the
desirable effects of Arg supplementation observed on overall growth performance and
feed efficiency in birds.
Excessive supplementation of Arg, at levels higher than the recommended, has shown
undesirable effects on the physiology and morphology of birds. Provision of Arg at
167 mg/L through drinking water resulted in the congestion of the vascular vessels in
the liver and kidneys in birds as compared with those receiving no supplementary L- Arg.
However, birds supplemented with 334 mg/L Arg exhibited destruction of the tubules
and glomeruli in the kidney and periportal mononuclear inflammatory infiltration in the
liver possibly due to a high production of NO (Alabi et al. 2018). Dietary supplementa­
tion of different digestible Arg: Lys ratios (1.15, 1.15, 1.16 and 1.171) did not influence the
meat quality attributes, foot pad dermatitis (FPD) occurrence, and breast myopathies in
broilers (Zampiga et al. 2018).

Effect of Arg supplementation on immune response and related physiological

parameters
Arginine is a metabolically versatile amino acid that can influence the immune function
of birds under challenging conditions. A group of enzymes (NO synthases) is responsible
for the conversion of Arg into citrulline and NO (Förstermann et al. 1991), as a sole route
for NO synthesis (Fernandes and Murakami 2010). Nitric oxide performs the crucial
function of cytotoxic effects on immune activated cells and regulates immune function
along with several other important functions in the body (Hibbs et al. 1988). In cocci­
diosis challenged broilers, depletion of Arg was observed due to higher expression of
inducible NO synthase (iNOS) to reduce the Eimeria proliferation (Tan et al. 2014a).
Reduction in dietary Arg (40%) resulted in decreased NO levels in the plasma of healthy
birds, but it was not observed in the infected birds, indicating high metabolic prioritisa­
tion of Arg for the synthesis of NO during coccidiosis (Rochell et al. 2017).
Many studies have explored the immunomodulatory effects of Arg in broilers (Table
3). Improvement in immune response after a Salmonella Typhimurium challenge in
young broiler chicks, in response to supplementation of Arg and vitamin E together, has
been reported (Liu et al. 2014). This combination also improved bacterial resistance
against other pathogens in commercial growing conditions. As stated above, in ovo
feeding of Arg solution is the most effective technology for regulating early nutrient
supply and immune function in birds (Gao et al. 2017b). The main immune-modulatory
effects of Arg are attributed to its ability to synthesise NO as many studies have reported
an increase in the levels of NO in birds following supplementation with Arg (Ruiz-Feria
2009). Arginine has shown synergistic effects when supplemented with other antioxi­
dants like vitamin C and E. For example, 1% Arg along with vitamin C and E resulted in
increased NO synthesis and decreased pulmonary hypertension (Ruiz-Feria and
Abdukalykova 2009). However, supplementation of Arg alone at 1.21% also resulted in
higher NO levels in broilers (Basoo et al. 2012). Arginine supplementation is particularly
 WORLD’S POULTRY SCIENCE JOURNAL 527

useful in poultry farming at higher altitudes as it can maintain the plasma NO level to
relax vascular smooth muscle and lower the resistance to pulmonary blood flow
(Wideman et al. 2007). Moreover, Arg also alleviated the adverse effects of cold tem­
perature by reduction in the right ventricle to total ventricle ratio and blood haematocrit
at the 35th day of age in broilers (Kodambashi Emami et al. 2017).
Arginine increased the synthesis of interleukin and proliferation of T lymphocytes in
broilers (Yeh et al. 2002; Wu et al. 2009). Moreover, Arg can significantly affect
haematological parameters in birds indicating a crucial role in metabolic and physio­
logical haemostasis of the body (Figure 4). Birds fed diets containing Arg at 8.5 mg/kg
had lower triglyceride and cholesterol levels (Yang et al. 2016). Supplementation of Arg
at 167 mg/L of drinking water in Shika Brown chickens significantly improved hae­
moglobin, red blood cell, and packed cell volume (Animashahun et al. 2018). In ovo
injection (100 µg/100 µL/egg) of Arg exhibited a desirable effect on serum glutamate
pyruvate transaminase (SGPT), serum glutamate oxaloacetate transaminase (SGOT),
myoblast determination protein, and IgM in chicken embryos (Subramaniyan et al.
2019). Supplementation of 2 times tryptophan and 2.5 times more Arg than the NRC
recommendation resulted in higher serum concentration of glucose, total protein,
albumin, and creatine kinase in broilers (Emadi et al. 2011). Furthermore, in-ovo
injection of Arg and threonine improved antibody titre against sheep red blood cells
(SRBC) although no improvement was observed against influenza and Newcastle
disease viruses (Toghyani et al. 2019). However, Arg injected alone increased bursa
and spleen relative weights in birds (Toghyani et al. 2019). In the presence of
a sufficient level of dietary methionine, Arg supplementation significantly improved
immune function by elevating the antibody titres against infectious bronchitis and
infectious bursal disease viruses while decreasing serum uric acid levels (Jahanian and

Figure 4. Overall benefits of Arg supplementation in broiler chicken.

528 F. HASSAN ET AL.

Khalifeh-Gholi 2018). These findings indicate potential health-promoting effects of Arg

and its synergism with other amino acids in the bird’s diet. Arginine has also shown
synergism with trace minerals like Mn. A combination of Mn-amino acid complex and
Arg (Dig. 1.12 or 1.20%) resulted in higher levels of IgM in Salmonella Enteritidis
challenged broilers (Burin et al. 2019).
In ovo injection of 1% Arg solution in broiler breeders resulted in a significant
increase in the concentration of ghrelin and glucagon-like peptide 2 (Gao et al. 2017a);
it also improved inducible iNOS, contents of interleukin-4 (IL-4), NO and IgA, as well
as the weights of lymphoid organs and immunological barrier functions in the intest­
inal mucosa (Gao et al. 2017b). Arginine supplementation resulted in a quadratic
decrease (P < 0.05) in the expression of interleukin-1b (IL-1b), IL-18 mRNA and
interferon-γ (IFN-γ) mRNA in the spleen (Hu et al. 2016). On the other hand, increase
in dietary Arg linearly enhanced the expression of lymphotactin mRNA and toll-like
receptor 15 (TLR15) mRNA and macrophage inflammatory protein-1β (MIP-1β) in the
caecal tonsils. Moreover, Arg also linearly increased the activities of glutathione
peroxidase, catalase, and lysozyme in the liver. However, the liver malondialdehyde
and superoxide dismutase activities were not affected by the Arg supplementation (Hu
et al. 2016). Arginine suppressed the TLR 4 pathway and reduced the CD14+ cell
percentage in the spleen to attenuate systemic inflammation while reducing pro-
inflammatory cytokines in lipopolysaccharide challenged broilers (Tan et al. 2014b).
Arginine improved the immune response and modulated the circulating T cells to
potentially alleviate the immunosuppression caused by vaccination against infectious
bursal disease (Tan et al. 2015). Bortoluzzi, Rochell, and Applegate (2017) have
extensively reviewed the potential effects of essential amino acids including Arg on
the gut development, health, and integrity of intestinal mucosa and activation of the
innate immune system in broiler chickens.
Generally, Arg supplementation has been shown to increase the immunoglobulin in
birds, but very high levels (above 14.7 g/kg of diet) linearly decrease the serum IgM
and IgG contents with increasing Arg levels (Liu et al. 2019). Thus, broilers offered
19.1 or 23.4 g/kg Arg had lower serum IgG or IgM than those fed 9.8 g/kg Arg, which
was mainly attributed to the downregulation of mRNA expression of bursal IgM and
IgG. Supplementation of Arg deficient diets with Arg (0.45, 0.90, 1.35, and 1.80%)
resulted in higher serum concentrations of IGF-I, insulin, IgA, and IFN-γ as well as
high thymus weight in broiler chickens (Xu et al. 2018). Moreover, it also showed
linear and quadratic improvement in lymphocyte proliferation and antibody titres
against Newcastle disease. These findings indicate immune-modulatory effects of Arg
supplementation through mediating different immune-related pathways including pro
and anti-inflammatory cytokines.

Effect of Arg supplementation on gut health

Arginine is a major precursor in the body of polyamines, which play a crucial role in
intestinal development especially during early age (Loser et al. 1999). This may be the
main reason behind the positive effects of dietary Arg on the intestinal morphology of
broiler chicks (Murakami et al. 2012). Polyamines are responsible for cellular prolifera­
tion, migration, and apoptosis in the intestine (Ruemmele et al. 1999). As a precursor of
 WORLD’S POULTRY SCIENCE JOURNAL 529

polyamines, Arg mediates the trophic effects on mitosis in the intestinal region to
subsequently enhance villus size and number (Uni, Ganot, and Sklan 1998). Dietary
Arg linearly increased the mucosal density, which may be attributed to the indirect effect
of polyamines or direct effects of Arg on goblet cells or enterocyte replication; however, it
is not fully clear yet (Tan et al. 2014a). Tan et al. (2014a) reported potential of L-Arg to
reduce the mucosal disruption of intestine possibly through mediating TLR4 and mTOR
complex 1 pathways in broilers challenged with coccidiosis.
Arginine also affects the development and morphology of the gastrointestinal tract
(GIT) in chickens (Table 4). Being an essential amino acid, dietary Arg has been used to
improve the development of GIT in chicken (Yuan et al. 2015; Gao et al. 2018; Jha et al.
2019). Dietary supplementation of Arg at 1.94 and 2.19% levels improved the length and
relative weight of the small intestine, respectively (Yu et al. 2018b). Moreover, it also
affected the villus height of the ileum and morphology of duodenal mucosa in chicks.
Boilers fed diets with 0.2% Arg had a higher proportion of duodenum and jejunum
(Youvalari et al. 2017). Arginine supplementation also increased the jejunal villus surface
area (Kodambashi Emami et al. 2017). It also significantly improved the villi height to
crypt depth ratio in the intestine of Eimeria challenged birds fed with a basal diet
supplemented with 100, 105, and 110% of the NRC recommendations of Arg (Laika
and Jahanian 2017).
Dietary supplementation of L-Arg has been shown to protect the integrity of mucosal
membranes of the GIT by enhancing the innate immune response, nutrient absorption
and gut barrier function while reducing the intestinal colonisation of Clostridium per­
fringens in necrotic enteritis-challenged broilers (Zhang et al. 2017). Moreover, dietary L-
Arg (0.3%) relieved the gut injury and normalised the ileal microbiota of Clostridium
perfringens-challenged broilers. Activities of digestive enzymes, sucrose, maltase, and
alkaline phosphatase, in the jejunum were increased by in ovo feeding of 1% Arg solution.
It also increased the mRNA expressions of jejunal sensing receptors like taste receptor
type 1 (members 1 and 3), and G protein-coupled receptor class C, group 6, subtype
A (GPRC6A) in broilers (Gao et al. 2017a). These receptors are found in the cell
membrane, where they recognise substances, thereby functioning as mediators of signals
across the cellular membranes and respond to physiologically important substances such
as hormones and neurotransmitters (Clemmensen et al. 2014). GPRC6A is also involved
in regulation of inflammation, metabolism, and endocrine functions. A recent study has
shown that GPRC6A can directly regulate hepatic metabolism to control systemic energy
homoeostasis (Pi et al. 2020).
Furthermore, in-ovo supplementation of Arg also significantly increased the
activity of iNOS, IL-4, IL-2, and secretory immunoglobulin A (sIgA) contents.
Moreover, it substantially enhanced the mRNA expressions of TLR-4 and TLR-2
in the intestinal mucosa (Gao et al. 2017b). Gut health is a key for the optimum
performance of birds as it directly affects the absorption and utilisation of dietary
nutrients along with a crucial role in the immune response. Substantial evidence
regarding modulation of gut development, morphology and health advocate the
optimum supplementation of dietary Arg in poultry. Moreover, Arg is an ideal
candidate to be used for the early nutrition programming of birds as a strategy to
modulate gut health and immune response to harvest benefits of better health and
production in the commercial poultry industry.
 530
F. HASSAN ET AL.

Table 4. Effect of Arg supplementation on gut health of broiler.

Reference Level Used Results
Khajali et al. (2011) 0, 0.2 and 0.4% Villus height, width and surface area of duodenum, jejenum and ileum were not affected by Arg supplementation
Murakami et al. (2012) 1.39, 1.49, 1.59, 1.69 and 1.79% Small intestine length and weight was not affected by treatments
Tan et al. (2014a) 11.1, 13.3 and 20.2 g/kg Higher Arg concentration increased jejunal villus height and linearly increased jejunal crypt depth
Fernandes et al. (2009) 0 and 1% Arg supplementation had higher small intestine length and crypt depth of duodenum. However, small intestine weight and
villus height were not affected by treatments
Laika and Jahanian 100, 105 and 110% Improved the villi height to crypt depth ratio in intestine
(2017)
Gao et al. (2017a) 0.5, 1 or 2% Some mRNA expressions of jejunal sensing receptors and activities of digestive enzymes, sucrose, maltase and alkaline
phosphatase, in jejunum were increased
Gao et al. (2017b) 1% Activity of iNOS, contents of IL-4 and IL-2) and sIgA, mRNA expressions of toll-like receptor-4 and toll-like receptor-2 in
intestinal mucosa were increased
Youvalari et al. (2017) CuSO4 (0, 125 & 250 ppm) and Arg (0, Enhanced the proportion of duodenum and jejunum
0.1 and 0.2%)
Kodambashi Emami GAA as 0.6 or 1.2 &/or Arg as 0.86 or Higher jejunal villus surface area
et al. (2017) 1.72 g/Kg
Gao et al. (2018) 1% In ovo feeding of L- Arg increased villus height and decreased crypt depth in duodenum of broiler embryos and post-hatch
hatchlings
Yu et al. (2018b) 1.19, 1.44, 1.69, 1.94 or 2.19% Length and relative weight of the small intestine, villus height of the ileum and morphology of duodenal mucosa were
improved
 WORLD’S POULTRY SCIENCE JOURNAL 531

Conclusion
Arginine is an important essential amino acid with a major role in the growth and
immune functions of birds due to its ability to produce NO, creatine, and polyamines.
Being a trophic amino acid, Arg possesses an excellent ability to modulate gut develop­
ment and enhance intestinal recovery after infection by substantially enhancing epithelial
turnover rates. Moreover, L-Arg is an ideal candidate for early nutritional programming
(in ovo and post-hatch) in poultry to enhance gut development and to improve overall
health and performance of poultry. It is concluded that optimum dietary supplementa­
tion of L-Arg above NRC recommended levels should be considered in broiler produc­
tion keeping in view the physiological stage and immune status of the birds.

Acknowledgments
All the authours aknowledge the help of Mr. Khobaib Hameed regarding literature search and data
collection.

Disclosure of potential conflicts of interest

No potential conflict of interest was reported by the author(s).

Notes on contributors
Dr. F. Hassan, received his BSc. (Hons) Animal Husbandry in 2001 from University of
Agriculture, Faisalabad with distinction (Gold Medal). After that he secured his MSc (Hons) in
2004 and PhD degree in 2012 in the field of Animal Breeding and Genetics from same university.
Meanwhile, He joined the University of Agriculture, Faisalabad as Lecturer in July, 2004 and then
promoted as Assistant Professor in 2019. Dr. Hassan has published more than 40 articles in peer
reviewed well known international journals. Recently, Dr Hassan has completed two-year post­
doctrate (2018-2020) under ‘Talented Young Scientist’ fellowship of the Ministry of Science and
Technology, China. Dr Hassan has performed excellent research work on poultry nutrition,
breeding and genetic improvement. He has worked as a research scientist in two mega projects
focusing on development of indigenous chicken breeds in Pakistan to improve performance of
rural poultry and their genetic conservation. Moreover, Dr. Hassan has done research on mole­
cular nutrition of birds with a focus to understand the molecular effects of nutrients on different
physiological networks and expression of major genes.
Mr. M. A. Arshad received his M.Sc. (Hons) in Animal Nutrition from University of Agriculture
Faisalabad (Pakistan) in 2019. His key areas of research are poultry nutrition and economical
feeding strategies in broiler chicken. He has performed the studies regarding the supplementation
of bile acids and lipase enzyme and and their subsequent effects on the digestibility and meat
quality of the broiler chicken. From 2019 until present, he is working as Production Manager at
Haji Sharif Feeds, Faisalabad, Pakistan and supervises the feed formulation and quality control
departments.
Mr. S. Hassan received his MSc.(Hons) in Animal Nutrition from University of Agriculture
Faisalabad Pakistan in 2016. His research interests mainly involve dietary supplementation of
trace minerals in broiler diets to get optimum performance of birds. Moreover, he is working on
his PhD dissertation focusing the role of nano-particles of trace elements in poultry nutrition.
Dr. R. M. Bilal received his M.Sc (Hons) Animal Nutrition from the University of Agriculture
Faisalabad Pakistan in 2000 and later on earned his PhD in Poultry Nutrition from the same
532 F. HASSAN ET AL.

University in 2016 with a major work on layer phase feeding. Currently he is working as a Lecturer
in Animal Nutrition in Islamia University Bahawalpur, Pakistan. His main research interests are
the 1) Search of new feed stuffs instead of traditional conventional sources 2) gut health of boiler 3)
Immunity of poultry birds. He has produced more than ten research papers on different aspects of
poultry science and nutrition.
Dr. M. Saeed is presently working as Assistant Professor in the Department of Poultry Science,
Faculty of Animal Production & Technology, Cholistan University of Veterinary and Animal
Sciences, Bahawalpur –Pakistan. He secured two Silver Medals in Bachelor and M.Phil.
Dr. Muhammad Saeed has earned PhD in Poultry Nutrition with distinction (Chinese
Government award) in 2018 from North West Agriculture and Forestry University, Yangling,
Shaanxi China. In his Doctorate studies, he studied the role of L-Theanine dietary supplementa­
tion in avian nutrition and his PhD work he published peer reviewed research papers in well-
known International scientific journals (International Journal of Molecular Sciences, SCI 4.02 IF,
Poultry Science, SCI 2.02IF and Biomedicine & Pharmacotherapy SCI 3.7IF). Dr. Muhammad
Saeed has also presented his scientific work in different international conferences worldwide
including UK, Italy, Germany, Thailand, Malaysia and China. His research interests mainly
include phytogenic additives in Avian Nutrition with special emphasis on using herbs as anti­
biotics replacer in poultry Industry.
M. S. Rehman received his BSc. (Hons) Animal Husbandry in 2000 from University of
Agriculture, Faisalabad with distinction (Bronze Medal). Dr. Rehman secured his PhD degree in
Animal Breeding and Genetics in 2009 from the University of Agriculture, Faisalabad,
PAKISTAN. Meanwhile, He joined the University of Agriculture, Faisalabad as Lecturer in 2002
and then promoted as Assistant Professor in 2009. Dr. Rehman has published more than 25 articles
in peer reviewed well known international journals. Dr Rehman joined the Iowa State University
of Science and Technology, Ames, Iowa (USA) as a postdoctoral fellow during 2011-2012.
Dr Rehman has performed excellent research work on various aspects of animal nutrition,
breeding and genetic characterisation of farm animals. Moreover, Dr. Rehman has done research
on the effect of different feed additives on performance of ruminants and poultry.

ORCID
F. Hassan http://orcid.org/0000-0002-8781-0510
M. Saeed http://orcid.org/0000-0001-5048-5753

References
Abdukalykova, S., and C. A. Ruiz-Feria. 2006. “Arginine and Vitamin E Improve the Cellular and
Humoral Immune Response of Broiler Chickens.” International Journal of Poultry Science 5 (2):
121–127. doi:10.3923/ijps.2006.121.127.
Alabi, O., A. Shoyombo, R. Animashahun, S. Olawoye, J. Abdulazeez, O. Faduhunsi, and
D. Oladehinbo. 2018. “Effects of L-Arginine Supplementation of Drinking Water on the Kidney
and Liver of Sasso Chickens.” International Journal of Livestock Production 9 (7): 160–164.
doi:10.5897/IJLP2018.0472.
Al-Daraji, H. J., and A. M. Salih. 2012. “Effect of Dietary L-arginine on Productive Performance of
Broiler Chickens.” Pakistan Journal of Nutrition 11 (3): 252–257. doi:10.3923/pjn.2012.252.257.
Animashahun, R., O. Alabi, A. Shoyombo, M. Ayeni, A. Adesina, and A. Bolaji. 2018. “Effect of
Oral Supplementation of L-Arginine on the Hematological Indices of Shika Brown Chickens.”
In Proceeding of 43rd Annual Conference of the Nigerian Society for Animal Production, FUT
Owerri, March 18th – 22nd.
 WORLD’S POULTRY SCIENCE JOURNAL 533

Ball, R. O., K. L. Urschel, and P. B. Pencharz. 2007. “Nutritional Consequences of Interspecies

Differences in Arginine and Lysine Metabolism.” The Journal of Nutrition 137 (6): 1626S–1641S.
doi:10.1093/jn/137.6.1626S.
Balnave, D., and J. Brake. 2001. “Different Responses of Broilers at Low, High, or Cyclic
Moderate-high Temperatures to Dietary Sodium Bicarbonate Supplementation Due to
Differences in Dietary Formulation.” Australian Journal of Agricultural Research 52 (6):
609–613. doi:10.1071/AR00136.
Basoo, H., F. Khajali, E. A. Khoshoui, M. Faraji, and R. F. Wideman. 2012. “Re-evaluation of
Arginine Requirements for Broilers Exposed to Hypobaric Condition during the 3-to 6-week
Period.” The Journal of Poultry Science 49 (4): 303–307. doi:10.2141/jpsa.0110133.
Becker, W. A., J. V. Spencer, L. W. Mirosh, and J. A. Verstrate. 1979. “Prediction of Fat and Fat
Free Live Weight in Broiler Chickens Using Backskin Fat, Abdominal Fat, and Live Body
Weight.” Poultry Science 58 (4): 835–842. doi:10.3382/ps.0580835.
Birmani, W., A. Raza, A. Nawab, S. Tang, M. Waseem, G. Li, M. Xiao, and L. An. 2019.
“Importance of Arginine as Immune Regulator in Animal Nutrition.” International Journal of
Veterinary Sciences Research 5 (1): 1–10. doi:10.18488/journal.110.2019.51.1.10.
Bortoluzzi, C., S. Rochell, and T. Applegate. 2017. “Threonine, Arginine, and Glutamine:
Influences on Intestinal Physiology, Immunology, and Microbiology in Broilers.” Poultry
Science 97 (3): 937–945. doi:10.3382/ps/pex394.
Brake, J. 1998. “Optimum Dietary Arginine: Lysine Ratio for Broiler Chickens Is Altered during
Heat Stress in Association with Changes in Intestinal Uptake and Dietary Sodium Chloride.”
British Poultry Science 39 (5): 639–647. doi:10.1080/00071669888511.
Brake, J., and D. Balnave. 1995. “Essentiality of Arginine in Broilers during Hot Weather.” In
Proceedings of 12th Annual Biokyowa Amino Acid Council Meeting, St Louis, Mo,
October 3–5.
Bulbul, T., Z. Bozkurt, E. Ulutas, O. Yilmaz, and A. Bulbul. 2013. “The Effect of L-arginine on
Growth Performance, Some Serum Biochemical Parameters and Duodenal Motility in Broilers.”
Kafkas Universitesi Veteriner Fakultesi Dergisi 19 (5): 821–827.
Burin, A. M., Jr., N. L. M. Fernandes, A. Snak, A. Fireman, D. Horn, and J. I. M. Fernandes. 2019.
“Arginine and Manganese Supplementation on the Immune Competence of Broilers Immune
Stimulated with Vaccine against Salmonella Enteritidis.” Poultry Science 98 (5): 2160–2168.
doi:10.3382/ps/pey570.
Castro, F. L. S., and W. K. Kim. 2020. “Secondary Functions of Arginine and Sulfur Amino Acids
in Poultry Health.” Animals 10 (11): 2106. doi:10.3390/ani10112106.
Castro, F. L. S., S. Su, H. Choi, E. Koo, and W. K. Kim. 2019. “L-Arginine Supplementation
Enhances Growth Performance, Lean Muscle, and Bone Density but Not Fat in Broiler
Chickens.” Poultry Science 98 (4): 1716–1722. doi:10.3382/ps/pey504.
Castro, F. L. S., P. Y. Teng, S. Yadav, R. L. Gould, S. Craig, R. Pazdro, and W. K. Kim. 2020. “The
Effects of L-Arginine Supplementation on Growth Performance and Intestinal Health of Broiler
Chickens Challenged with Eimeria Spp.” Poultry Science 99 (11): 5844–5857. doi:10.1016/j.
psj.2020.08.017.
Chamruspollert, M., G. Pesti, and R. Bakalli. 2002. “Dietary Interrelationships among Arginine,
Methionine, and Lysine in Young Broiler Chicks.” British Journal of Nutrition 88 (6): 655–660.
doi:10.1079/BJN2002732.
Clemmensen, C., S. Smajilovic, P. Wellendorph, and H. Bräuner-Osborne. 2014. “The GPCR,
Class C, Group 6, Subtype A (GPRC6A) Receptor: From Cloning to Physiological Function.”
British Journal of Pharmacology 171 (5): 1129–1141. doi:10.1111/bph.12365.
Cuca, G. M., and L. Jensen. 1990. “Arginine Requirement of Starting Broiler Chicks.” Poultry
Science 69 (8): 1377–1382. doi:10.3382/ps.0691377.
Emadi, M., F. Jahanshiri, K. Kaveh, M. Hair-Bejo, A. Ideris, and A. Alimon. 2011. “Nutrition and
Immunity: The Effects of the Combination of Arginine and Tryptophan on Growth
Performance, Serum Parameters and Immune Response in Broiler Chickens Challenged with
Infectious Bursal Disease Vaccine.” Avian Pathology 40 (1): 63–72. doi:10.1080/
03079457.2010.539590.
534 F. HASSAN ET AL.

Fernandes, J., A. Murakami, E. Martins, M. Sakamoto, and E. Garcia. 2009. “Effect of Arginine on
the Development of the Pectoralis Muscle and the Diameter and the Protein: Deoxyribonucleic
Acid Rate of Its Skeletal Myofibers in Broilers.” Poultry Science 88 (7): 1399–1406. doi:10.3382/
ps.2008-00214.
Fernandes, J. I. M., and A. E. Murakami. 2010. “Arginine Metabolism in Uricotelic Species.” Acta
Scientiarum Animal Sciences 32 (4): 357–366. doi:10.4025/actascianimsci.v32i4.10990.
Förstermann, U., H. H. Schmidt, J. S. Pollock, H. Sheng, J. A. Mitchell, T. D. Warner, M. Nakane,
and F. Murad. 1991. “Isoforms of Nitric Oxide Synthase Characterization and Purification from
Different Cell Types.” Biochemical Pharmacology 42 (10): 1849–1857. doi:10.1016/0006-
2952(91)90581-O.
Fouad, A., H. El-Senousey, X. Yang, and J. Yao. 2012. “Role of Dietary L-arginine in Poultry
Production.” International Journal of Poultry Science 11 (11): 718–729. doi:10.3923/
ijps.2012.718.729.
Gao, T., M. Zhao, Y. Li, L. Zhang, J. Li, L. Yu, F. Gao, and G. Zhou. 2018. “Effects of in Ovo Feeding
of L-arginine on the Development of Digestive Organs, Intestinal Function and Post-hatch
Performance of Broiler Embryos and Hatchlings.” Journal of Animal Physiology and Animal
Nutrition 102 (1): e166–e175. doi:10.1111/jpn.12724.
Gao, T., M. Zhao, L. Zhang, J. Li, L. Yu, P. Lv, F. Gao, and G. Zhou. 2017a. “Effect of in Ovo
Feeding of L-arginine on the Hatchability, Growth Performance, Gastrointestinal Hormones,
and Jejunal Digestive and Absorptive Capacity of Posthatch Broilers.” Journal of Animal Science
95 (7): 3079–3092.
Gao, T., M. Zhao, L. Zhang, J. Li, L. Yu, P. Lv, F. Gao, and G. Zhou. 2017b. “Effects of in Ovo
Feeding of L-arginine on the Development of Lymphoid Organs and Small Intestinal Immune
Barrier Function in Posthatch Broilers.” Animal Feed Science and Technology 225: 8–19.
doi:10.1016/j.anifeedsci.2017.01.004.
Guo, Y. W., B. L. Shi, S. M. Yan, Y. Q. Xu, J. L. Li, and T. Y. Li. 2015. “Effects of Arginine on
Cytokines and Nitric Oxide Synthesis in Broilers.” Journal of Animal and Plant Sciences 25 (2):
366–371.
Hibbs, J. B., Jr, R. R. Taintor, Z. Vavrin, and E. M. Rachlin. 1988. “Nitric Oxide: A Cytotoxic
Activated Macrophage Effector Molecule.” Biochemical and Biophysical Research
Communications 157 (1): 87–94. doi:10.1016/S0006-291X(88)80015-9.
Hu, Y.-D., J.-Z. Tan, Q. Ji, and H.-F. Zhang. 2016. “Regulatory Effects of Dietary L-Arg
Supplementation on the Innate Immunity and Antioxidant Ability in Broiler Chickens.”
Journal of Integrative Agriculture 15 (11): 2578–2587. doi:10.1016/S2095-3119(16)61404-1.
Jahanian, R. 2009. “Immunological Responses as Affected by Dietary Protein and Arginine
Concentrations in Starting Broiler Chicks.” Poultry Science 88 (9): 1818–1824. doi:10.3382/
ps.2008-00386.
Jahanian, R., and M. Khalifeh-Gholi. 2018. “Marginal Deficiencies of Dietary Arginine and
Methionine Could Suppress Growth Performance and Immunological Responses in Broiler
Chickens.” Journal of Animal Physiology and Animal Nutrition 102 (1): e11–e20. doi:10.1111/
jpn.12695.
Jha, R., A. K. Singh, S. Yadav, J. F. D. Berrocoso, and B. Mishra. 2019. “Early Nutrition
Programming (In Ovo and Post-hatch Feeding) as a Strategy to Modulate Gut Health of
Poultry.” Frontiers in Veterinary Science 6: 82. doi:10.3389/fvets.2019.00082.
Khajali, F., M. Faraji, and H. Basoo. 2013. “Estimation of Arginine Requirements for Male Broilers
Grown at High Altitude from One to Twenty-one Days of Age.” Journal of Agricultural Science
Technology 15 (5): 911–917.
Khajali, F., M. Tahmasebi, H. Hassanpour, M. Akbari, D. Qujeq, and R. Wideman. 2011. “Effects of
Supplementation of Canola Meal-based Diets with Arginine on Performance, Plasma Nitric
Oxide, and Carcass Characteristics of Broiler Chickens Grown at High Altitude.” Poultry Science
90 (10): 2287–2294. doi:10.3382/ps.2011-01618.
Khajali, F., and R. Wideman. 2010. “Dietary Arginine: Metabolic, Environmental, Immunological
and Physiological Interrelationships.” World’s Poultry Science Journal 66 (4): 751–766.
doi:10.1017/S0043933910000711.
 WORLD’S POULTRY SCIENCE JOURNAL 535

Kidd, M., E. Peebles, S. Whitmarsh, J. Yeatman, and R. Wideman Jr. 2001. “Growth and Immunity
of Broiler Chicks as Affected by Dietary Arginine.” Poultry Science 80 (11): 1535–1542.
doi:10.1093/ps/80.11.1535.
Kodambashi Emami, N., A. Golian, D. Rhoads, and M. Danesh Mesgaran. 2017. “Interactive
Effects of Temperature and Dietary Supplementation of Arginine or Guanidinoacetic Acid on
Nutritional and Physiological Responses in Male Broiler Chickens.” British Poultry Science 58
(1): 87–94. doi:10.1080/00071668.2016.1257779.
Kwak, H., R. Austic, and R. Dietert. 1999. “Influence of Dietary Arginine Concentration on
Lymphoid Organ Growth in Chickens.” Poultry Science 78 (11): 1536–1541. doi:10.1093/ps/
78.11.1536.
Labadan, M., Jr, K.-N. Hsu, and R. Austic. 2001. “Lysine and Arginine Requirements of Broiler
Chickens at Two-to Three-week Intervals to Eight Weeks of Age.” Poultry Science 80 (5):
599–606. doi:10.1093/ps/80.5.599.
Laika, M., and R. Jahanian. 2017. “Increase in Dietary Arginine Level Could Ameliorate
Detrimental Impacts of Coccidial Infection in Broiler Chickens.” Livestock Science 195: 38–44.
doi:10.1016/j.livsci.2016.11.002.
Li, P., Y. L. Yin, D. Li, S. W. Kim, and G. Wu. 2007. “Amino Acids and Immune Function.” British
Journal of Nutrition 98 (2): 237–252. doi:10.1017/S000711450769936X.
Liu, S., J. Tan, Y. Hu, X. Jia, M. H. Kogut, J. Yuan, and H. Zhang. 2019. “Dietary L-arginine
Supplementation Influences Growth Performance and B-cell Secretion of Immunoglobulin in
Broiler Chickens.” Journal of Animal Physiology and Animal Nutrition 103 (4): 1125–1134.
doi:10.1111/jpn.13110.
Liu, X., J. Byrd, M. Farnell, and C. Ruiz-Feria. 2014. “Arginine and Vitamin E Improve the
Immune Response after a Salmonella Challenge in Broiler Chicks.” Poultry Science 93 (4):
882–890. doi:10.3382/ps.2013-03723.
Loser, C., A. Eisel, D. Harms, and U. Fölsch. 1999. “Dietary Polyamines are Essential Luminal
Growth Factors for Small Intestinal and Colonic Mucosal Growth and Development.” Gut 44
(1): 12–16. doi:10.1136/gut.44.1.12.
Mejia, L., C. Zumwalt, P. Tillman, R. Shirley, and A. Corzo. 2012. “Ratio Needs of Arginine
Relative to Lysine of Male Broilers from 28 to 42 Days of Age during a Constant, Elevated
Environmental Temperature Regimen.” Journal of Applied Poultry Research 21 (2): 305–310.
doi:10.3382/japr.2011-00397.
Munir, K., M. Muneer, E. Masaoud, A. Tiwari, A. Mahmud, R. Chaudhry, and A. Rashid. 2009.
“Dietary Arginine Stimulates Humoral and Cell-mediated Immunity in Chickens Vaccinated
and Challenged against Hydropericardium Syndrome Virus.” Poultry Science 88 (8): 1629–1638.
doi:10.3382/ps.2009-00152.
Murakami, A. E., J. I. Fernandes, L. Hernandes, and T. C. Santos. 2012. “Effects of Starter Diet
Supplementation with Arginine on Broiler Production Performance and on Small Intestine
Morphometry.” Pesquisa Veterinaria Brasileira 32 (3): 259–266. doi:10.1590/S0100-
736X2012000300014.
Nayak, N., R. A. Rajini, S. Ezhilvalavan, A. R. Sahu, and J. J. Kirubaharan. 2016. “Influence of
In-ovo Arginine Feeding on Post-hatch Growth Performance and Economics of Broilers.”
Journal of Animal Research 6 (4): 585–591. doi:10.5958/2277-940X.2016.00067.X.
NRC. 1994. Requirements of Poultry. 9th ed. Washington, DC: National Academies Press.
Pi, M., F. Xu, R. Ye, S. K. Nishimoto, R. W. Williams, L. Lu, and L. Darryl Quarles. 2020. “”Role of
GPRC6A in Regulating Hepatic Energy Metabolism in Mice.”.” Scientific Reports 10 (1): 1–12.
doi:10.1038/s41598-020-64384-8.
Rochell, S., A. Helmbrecht, C. M. Parsons, and R. N. Dilger. 2017. “Interactive Effects of Dietary
Arginine and Eimeria Acervulina Infection on Broiler Growth Performance and Metabolism.”
Poultry Science 96 (3): 659–666. doi:10.3382/ps/pew295.
Rubin, L. L., C. W. Canal, A. Ribeiro, A. Kessler, I. Silva, L. Trevizan, T. Viola, M. Raber,
T. A. Gonçalves, and R. Krás. 2007. “Effects of Methionine and Arginine Dietary Levels on
the Immunity of Broiler Chickens Submitted to Immunological Stimuli.” Brazilian Journal of
Poultry Science 9 (4): 241–247. doi:10.1590/S1516-635X2007000400006.
536 F. HASSAN ET AL.

Rueda, E., C. Michelangeli, and F. Gonzalez-Mujica. 2003. “L-canavanine Inhibits L-arginine

Uptake by Broiler Chicken Intestinal Brush Border Membrane Vesicles.” British Poultry
Science 44 (4): 620–625. doi:10.1080/00071660310001618299.
Ruemmele, F., C. Ruemmele, E. Levy, and E. Seidman. 1999. “Molecular Mechanisms of the
Regulation of Intestinal Epithelial Cell Turnover by Nutrients.” Clinical and Biological
Gastroenterology 23 (1): 47–55.
Ruiz-Feria, C. 2009. “Concurrent Supplementation of ArgininE, Vitamin E, and Vitamin
C Improve Cardiopulmonary Performance in Broilers Chickens.” Poultry Science 88 (3):
526–535. doi:10.3382/ps.2008-00401.
Ruiz-Feria, C., and S. Abdukalykova. 2009. “Arginine and Vitamin E Improve the Antibody
Responses to Infectious Bursal Disease Virus (IBDV) and Sheep Red Blood Cells in Broiler
Chickens.” British Poultry Science 50 (3): 291–297. doi:10.1080/00071660902942759.
Sirathonpong, O., Y. Ruangpanit, O. Songserm, E. Koo, and S. Attamangkune. 2019.
“Determination of the Optimum Arginine: Lysine Ratio in Broiler Diets.” Animal Production
Science 59 (9): 1705–1710. doi:10.1071/AN18049.
Srinongkote, S., M. Smriga, and Y. Toride. 2004. “Diet Supplied with L-lysine and L-arginine
during Chronic Stress of High Stock Density Normalizes Growth of Broilers.” Animal Science
Journal 75 (4): 339–343. doi:10.1111/j.1740-0929.2004.00195.x.
Subramaniyan, S. A., D. R. Kang, J. R. Park, S. H. Siddiqui, P. Ravichandiran, D. J. Yoo, C. S. Na,
and K. S. Shim. 2019. “Effect of in Ovo Injection of L-Arginine in Different Chicken Embryonic
Development Stages on Post-Hatchability, Immune Response, and Myo-D and Myogenin
Proteins.” Animals 9 (6): 357. doi:10.3390/ani9060357.
Tamir, H., and S. Ratner. 1963a. “Enzymes of Arginine Metabolism in Chicks.” Archives of
Biochemistry and Biophysics 102 (2): 249–258. doi:10.1016/0003-9861(63)90178-4.
Tamir, H., and S. Ratner. 1963b. “A Study of Ornithine, Citrulline and Arginine Synthesis in
Growing Chicks.” Archives of Biochemistry and Biophysics 102 (2): 259–269. doi:10.1016/0003-
9861(63)90179-6.
Tan, J., T. Applegate, S. Liu, Y. Guo, and S. D. Eicher. 2014a. “Supplemental Dietary L-arginine
Attenuates Intestinal Mucosal Disruption during a Coccidial Vaccine Challenge in Broiler
Chickens.” British Journal of Nutrition 112 (7): 1098–1109. doi:10.1017/S0007114514001846.
Tan, J., Y. M. Guo, T. J. Applegate, E. C. Du, and X. Zhao. 2015. ““Dietary L-arginine Modulates
Immunosuppression in Broilers Inoculated with an Intermediate Strain of Infectious Bursa
Disease Virus.”.” Journal of the Science of Food and Agriculture 95 (1): 126–135. doi:10.1002/
jsfa.6692.
Tan, J., S. Liu, Y. Guo, T. J. Applegate, and S. D. Eicher. 2014b. “Dietary L-arginine
Supplementation Attenuates Lipopolysaccharide-induced Inflammatory Response in Broiler
Chickens.” British Journal of Nutrition 111 (8): 1394–1404. doi:10.1017/S0007114513003863.
Toghyani, M., S. Tahmasebi, M. Modaresi, and S. S. Ale Saheb Fosoul. 2019. “Effect of Arginine
and Threonine in Ovo Supplementation on Immune Responses and Some Serum Biochemical
Attributes in Broiler Chickens.” Italian Journal of Animal Science 18 (1): 342–349. doi:10.1080/
1828051X.2018.1529545.
Uni, Z., S. Ganot, and D. Sklan. 1998. “Posthatch Development of Mucosal Function in the Broiler
Small Intestine.” Poultry Science 77 (1): 75–82. doi:10.1093/ps/77.1.75.
Wideman, R., M. Chapman, K. Hamal, O. Bowen, A. Lorenzoni, G. Erf, and N. Anthony. 2007.
“An Inadequate Pulmonary Vascular Capacity and Susceptibility to Pulmonary Arterial
Hypertension in Broilers.” Poultry Science 86 (5): 984–998. doi:10.1093/ps/86.5.984.
Woodward, B. 1998. “Protein, Calories, and Immune Defenses.” Nutrition Reviews 56 (1): S84–
S92. doi:10.1111/j.1753-4887.1998.tb01649.x.
Wu, G., F. W. Bazer, T. A. Davis, S. W. Kim, P. Li, J. M. Rhoads, M. C. Satterfield, S. B. Smith,
T. E. Spencer, and Y. Yin. 2009. “Arginine Metabolism and Nutrition in Growth, Health and
Disease.” Amino Acids 37 (1): 153–168. doi:10.1007/s00726-008-0210-y.
Xu, Y., Y. Guo, B. Shi, S. Yan, and X. Guo. 2018. “Dietary Arginine Supplementation Enhances the
Growth Performance and Immune Status of Broiler Chickens.” Livestock Science 209: 8–13.
doi:10.1016/j.livsci.2018.01.001.
 WORLD’S POULTRY SCIENCE JOURNAL 537

Yang, H., X. Ju, Z. Wang, Z. Yang, J. Lu, and W. Wang. 2016. “Effects of Arginine Supplementation
on Organ Development, Egg Quality, Serum Biochemical Parameters, and Immune Status of
Laying Hens.” Brazilian Journal of Poultry Science 18 (1): 181–186. doi:10.1590/1516-
635x1801181-186.
Yeh, C. L., S. L. Yeh, M. T. Lin, and W. J. Chen. 2002. “Effects of Arginine-enriched Total
Parenteral Nutrition on Inflammatory-related Mediator and T-cell Population in Septic Rats.”
Nutrition 18 (7–8): 631–635. doi:10.1016/S0899-9007(02)00809-2.
Youvalari, S. A., P. Farhoomand, P. B. Kanani, and B. H. Ghasemabad. 2017. “Effects of Copper
Sulfate and Arginine Supplements on Performance and Carcass Traits in Broiler Chickens Fed
with Canola Meal Based Diet.” Iranian Journal of Applied Animal Science 7 (4): 647–654.
Yu, J., H. Yang, Z. Wang, H. Dai, L. Xu, and C. Ling. 2018b. “Effects of Arginine on the Growth
Performance, Hormones, Digestive Organ Development and Intestinal Morphology in the Early
Growth Stage of Layer Chickens.” Italian Journal of Animal Science 17 (4): 1077–1082.
doi:10.1080/1828051X.2018.1434692.
Yu, L. L., T. Gao, M. M. Zhao, P. A. Lv, L. Zhang, J. L. Li, Y. Jiang, F. Gao, and G. H. Zhou. 2018a.
“In Ovo Feeding of L-arginine Alters Energy Metabolism in Post-hatch Broilers.” Poultry
Science 97 (1): 140–148. doi:10.3382/ps/pex272.
Yuan, C., Y. Ding, Q. He, M. Azzam, J. Lu, and X. Zou. 2015. “L-arginine Upregulates the Gene
Expression of Target of Rapamycin Signaling Pathway and Stimulates Protein Synthesis in
Chicken Intestinal Epithelial Cells.” Poultry Science 94 (5): 1043–1051. doi:10.3382/ps/pev051.
Zampiga, M., L. Laghi, M. Petracci, C. Zhu, A. Meluzzi, S. Dridi, and F. Sirri. 2018. “Effect of
Dietary Arginine to Lysine Ratios on Productive Performance, Meat Quality, Plasma and
Muscle Metabolomics Profile in Fast-growing Broiler Chickens.” Journal of Animal Science
and Biotechnology 9 (1): 79. doi:10.1186/s40104-018-0294-5.
Zhang, B., Z. Lv, Z. Li, W. Wang, G. Li, and Y. Guo. 2017. “Dietaryl-arginine Inhibits Intestinal
Clostridium Perfringens Colonisation and Attenuates Intestinal Mucosal Injury in Broiler
Chickens.” British Journal of Nutrition 118 (5): 321–332. doi:10.1017/S0007114517002094.