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Effect of dietary phosphorus levels on meat quality and lipid metabolism in


broiler chickens
Xue-ke Li, Jin-zhi Wang, Chun-qing Wang, Chun-hui Zhang, Xia Li, Chunhong Tang, Xiu-li Wei
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http://dx.doi.org/10.1016/j.foodchem.2016.02.133
FOCH 18854

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Food Chemistry

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7 August 2015
4 February 2016
21 February 2016

Please cite this article as: Li, X-k., Wang, J-z., Wang, C-q., Zhang, C-h., Li, X., Tang, C-h., Wei, X-l., Effect of
dietary phosphorus levels on meat quality and lipid metabolism in broiler chickens, Food Chemistry (2016), doi:
http://dx.doi.org/10.1016/j.foodchem.2016.02.133

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Effect of dietary phosphorus levels on meat quality and lipid metabolism in

broiler chickens

Xue-ke Li a, b,1, Jin-zhi Wang a,1, Chun-qing Wang a, Chun-hui Zhang a*, Xia Li a,

Chun-hong Tang b, Xiu-li Wei a

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Agricultural Sciences, Beijing, China, 100193

Business University, Chongqing, China, 400067

Institute of Agro-Products Processing Science and Technology, Chinese Academy of

College of Environmental and Biological Engineering, Chongqing Technology and

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* Corresponding author

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Tel: +86 10 62819469

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Fax: +86 10 62815950

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E-mail: dr_zch@163.com (C. H. Zhang)

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Address: No. 2 Yuan Ming Yuan West Road, Haidian District, Beijing, China, 100193

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1. The authors contributed equally to the work

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Running Title: Dietary phosphorus level and meat quality in chicken

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Abstract

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To analyze the influence of dietary phosphorus (P) levels on meat quality and lipid

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metabolism, a 42-day feeding experiment (P deficient group; normal group; high P

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level groups of H1 and H2, respectively) using 100 one-day-old broilers was

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conducted. Results demonstrated that the quality of broiler chicken meat in deficient

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or high P groups decreased relative to the normal group. High P diets resulted in

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increased lightness, redness values, shear forces and decreased fatty acid contents and

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intramuscular fat content in breast meat (p < 0.01). Compared with normal group,

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lower malic enzyme activity, higher fatty acid synthase and AMP-activated protein

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kinase activities were observed in the treatment groups (p < 0.05). Chickens fed with

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normal diets had the lowest serum total cholesterol and triglyceride levels which

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differed from that of other treatments (p < 0.05). High-P diets significantly decreased

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the lipid accumulation in the liver (p < 0.01), whereas phosphorus levels in breast

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meat increased significantly (p < 0.01). It can be concluded that deficient or higher P

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levels could affect meat quality and expression of indicators on lipid metabolism of

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broiler chickens.

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Keywords: Phosphorus levels; Meat quality; Lipid metabolism; Broiler chickens.

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1. Introduction

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Chicken meat is considered a better choice for health diets because it contains less

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fat, as well as a higher proportion of polyunsaturated fatty acids (PUFA), when


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compared with meat from other species (Riovanto et al., 2012). Due to growing

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concerns on the relationship between diet and health, consumers are increasingly

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demanding high-quality meat products. Consequently, factors that affect meat quality

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in chickens are receiving the attention of both researchers and consumers.

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Meat quality in chickens is affected by various potential intrinsic and extrinsic

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factors. Among factors, phosphorus (P) is a critical and expensive mineral in poultry

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nutrition. This mineral is used in energy pathways and during the synthesis of cell

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membranes (Hill et al., 2008). P cannot be synthesized. It must be obtained from

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dietary sources (Weinera et al., 2001). Studies on effects of dietary P levels on animal

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growth and bone development showed that high P intake negatively impacts calcium

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metabolism and bone properties, whereas low P diets will limit the growth of the

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animals (Roman-Garcia et al., 2010). Currently, P recommendations appear to have

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exaggerated the availability of P in animals, and many countries are considering

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revisions to the P recommendations for broiler chicken feed (Yan et al., 2011). In

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addition, farmers tend to use feed with more P than the recommended level (Lopez et

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al., 2004). By far, there are barely any reports on the relationship between dietary P

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levels and meat quality.

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Moreover, P is a critical component during glycolysis, energy and lipid metabolism.

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All enzymes in the body need P (Takeda et al., 2012), but few reports describing how

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P affects lipid metabolism in broiler chickens are available. Lipid metabolism in

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chickens is affected by the activities of lipase and lipogenesis enzymes, including


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AMP-activated protein kinase (AMPK), lipase, and main lipogenic enzymes (fatty

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acid synthase (FAS), and malic enzyme (ME), etc. Among those enzymes, FAS can

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alter the rates of the biosynthesis and hydrolysis of fatty acids (Smith, Witkowski, &

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Joshi, 2003), and AMPK is primarily a critical regulator for energy metabolism (Kim

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et al., 2004), regulating numerous intracellular pathways such as fatty acid oxidation

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(Jonathan, John, & Bruce, 2012), development of fat cells and the deposition of

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intramuscular fatIMF (Hardie, 2007), and low AMPK activity is expected to

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enhance intramuscular fat (IMF) contents (Underwood et al., 2008). Therefore, we

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hypothesized that dietary P level may change the enzyme activities in broiler chickens,

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thus affecting meat quality through lipid metabolism, since fatty acid composition and

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biosynthesis play a role during the regulation for IMF contents (Camerona et al.,

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2000).

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To investigate the effects of dietary P levels on the meat quality and lipid

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metabolism of broiler chickens, a 42-day feeding experiment was performed using

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broilers. The chemical composition, meat color, cooking loss, tenderness, pH values,

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fatty acid profiles and IMF content were determined to evaluate the differences in

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meat quality affected by dietary P levels. Besides, to help understand how dietary P

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levels affect lipid metabolism in broiler chickens, the activities of enzymes including

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FAS, ME and AMPK in breast meat and liver were determined, trying to provide a

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foundation for understanding the relation of dietary P levels and meat quality.

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2. Materials and Methods


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2.1 Animals and diet composition

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One hundred one-day old female broilers (Arbor Acres) were randomly allocated to

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4 groups with 25 birds for each treatment. All birds were housed in metallic pens of

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identical size (1.4 m 0.7 m 0.4 m) with 5 chickens per pen and provided with 12 h

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light and dark cycles. The feeding experiments were conducted in animal testing

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grounds of the Institute of Agro-products Processing Science and Technology,

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Chinese Academy of Agricultural Sciences. The birds were allowed to consume food

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and water ad libitum.

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The experiment was divided into four groups according to available phosphorus

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(AP) concentration of the feed: D (P deficient group), N (normal diet group), H1 (high

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P group with available P 0.86% and 0.82%), and H2 (high P group with available P

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1.16% and 1.10%). The nutrient levels of N group were based on the NRC (1994)

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recommendations, and chickens were fed by basal diet. And on this basis we set

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deficient group (D) and adequate groups (H1 and H2) with different P content in the

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diet. The dietary P levels were mainly obtained by adding potassium dihydrogen

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phosphate (feed grade) to the feed. The birds were given a diet based on corn and

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soybean meal, and the diet included both starter and grower phases. The formulation

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and nutrient levels of the feed are presented in Table 1. Contents of Ca and TP were

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measured according to the AOAC (1935, 1996), while the others were calculated

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values.

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All chickens were weighed on a weekly basis, in the morning between 7:00 a.m and
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8:00 a.m before the feed is offered. Body weight gain (BWG), feed intake (FI) and

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feed conversion ratio (FCR) were calculated at the end of growth phase.

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At 42 days of age, blood samples were collected from veins under the wings in all

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live chickens. The serum was separated via centrifugation at 2500 g and 4 C for 15

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min. The chicken weights, feed intake and mortality are shown in Table 2. And all the

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live chickens were slaughtered and eviscerated. The liver and pancreas were removed,

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rinsed in 0.9 % NaCl and immediately stored in liquid nitrogen for biochemical

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analysis. The chickens were divided after evisceration. The whole breasts of each

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chicken were cut off, and all samples were stored at -80 C until further analysis.

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2.2 Basic chemical composition and phosphorus contents

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Before analysis, frozen samples were thawed at 4 C and homogenized with a

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Waring blender (7012G, Texas, USA). Each of the following chemical analyses

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involved a single sample from each chicken and experimental replication was across

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all chickens in a particular treatment. The basic chemical composition of the meat was

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determined according to the methods of AOAC (2000). The moisture contents were

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determined by oven-drying method at 110 C for 24 h (AOAC, 950.46B). The total

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protein content was determined using the Kjeldahl method (AOAC, 928.08), and the

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total ash content was determined by weight after heating the samples in muffle

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furnace at 550 C for 4 h (AOAC, 920.153).

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About 0.5 ~ 1.5 g of each homogeneous meat sample from each chicken was

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weighed into crucible before being dried for 1 ~ 2 h at 110 C. The rest of procedure

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follows that used to determine the total ash content. Afterwards, 5.0 mL of HCl was

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added to the cooled crucible, and all rinses were transferred though a filter into a

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volumetric flask. The same procedure was used for the blank. The TP of the breast

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meat was determined from the ash solution by measuring the yellow color developed

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through the vanadate-molybdate method at 430 nm (T6 UV-VIS spectrophotometer,

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Purkinje, Beijing, China) (AOAC, 1995). The phosphorus contents were expressed as

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P.

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2.3 Shear forces, cooking losses, meat color and pH values

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The frozen breast meat samples were thawed at 4 C for 24 h and cut into 1 cm 1

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cm 4 cm along the direction of the muscle fibers. The shear force values were

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measured on raw meat samples using a Warner Bratzler shear force device (TA-XT2i

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Plus, probe HDP/BSW, Stable Micro System Ltd., UK) according to the method of

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Wattanachant et al., (2004).

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Approximately 5 g of meat was weighed, wrapped in airtight polythene bag and

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cooked for 30 min at 80 C. Afterwards, the samples were cooled at room temperature

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and analyzed for cooking loss.

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Meat color attributes including lightness (L*), redness (a*) and yellowness (b*)

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were determined using a hand-held Minolta colorimeter (chroma meter CR-400,

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Konica Minolta Sensing Inc., Japan) standardized against a white calibration plate.

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The pH values were determined 24 h post-mortem using a Testo 205 pH meter


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(Testo Instrument Co. Ltd., Germany) equipped with an insertion electrode.

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2.4 Total cholesterol and triglyceride in serum and liver

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The serum total cholesterol (TC) and triglyceride (TG) levels were measured using

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commercial kits (GPO-POD; Applygen Technologies Inc., Beijing, China) with a

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HITACHI 7020 biochemistry analyzer (HITACHI Ltd., Tokyo, Japan).

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The liver lipid extractions were conducted according to the procedures developed

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by Lin et al., (2013). Briefly, the liver lipids were extracted using chloroform and

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methanol (2:1, v/v). The TG in the liver was measured using commercial kits with a

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HITACHI 7020 biochemistry analyzer.

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2.5 Intramuscular fat content and fatty acid profiles

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The IMF content was measured using the methods described by Folch et al., (1957).

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The total lipids were extracted in triplicates from each homogenized breast meat

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sample and used for IMF determination.

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The fatty acid composition in meat was determined by gas chromatography. Fatty

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acid methyl esters (FAME) were prepared by esterification with acetyl chloride in

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methanol. Approximately 0.5 g of each meat sample was dissolved in 6 mL of acetyl

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chloride in methanol in a tube that was tightly capped after filling with nitrogen. The

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tube was then shaken vigorously and treated in a water bath for 2 h at 80 C. The tube

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was washed using 3.0 mL sodium carbonate solution three times, and the resultant

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solution was then centrifuged at 5000 g for 5 min. The supernatant was transferred
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into auto sampler vial for subsequent GC analysis.

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Next, 1.0 mL of the diluted fatty acid methyl ester solution was analyzed with a

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Varian 450 Gas Chromatograph (Varian Inc., USA) equipped with a flame ionization

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detector (FID) and a split injector. The injector and detector in the machine were set to

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240 and 260 C, respectively. The helium carrier gas flow rate was 1.0 mL per min.

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The initial oven temperature was 190 C, which was held for 7.5 min before

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increasing at 3 C per min to 220 C. The final temperature was held for 2.5 min. In

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the injector, the split valve was set to 1:60. The fatty acid composition of the sample

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was calculated using the peak areas and methyl ester standard mixtures. The average

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amount of each fatty acid was used to calculate the sum of the saturated fatty acid

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(SFA), monounsaturated fatty acid (MUFA) and PUFA.

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2.6 Enzyme activity measurements

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2.6.1 Assays of AMPK activity

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The homogenates of breast meat and liver tissue were obtained according to the

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methods of Underwood et al., (2008) with minor modifications. Briefly, the samples

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(0.50 g) were homogenized in 2.5 mL of an ice-cold extraction buffer (0.25 M

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D-mannitol; 0.05 M Tris/HCl, pH 7.4; 1 mM EDTA; 1 mM EGTA; 1 mM DTT; 50

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mM NaF; 5 mM sodium pyrophosphate). The enzymatic activities were determined

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using the supernatant obtained after centrifuging the homogenate (12000 g for 5

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min at 4 C), and the differences between various samples were evaluated. The AMPK

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activity was measured using commercially available kits (CUSABIO Biotech Co., Ltd,
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Wuhan, China) according to the manufacturer's instructions.

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2.6.2 Assays of lipase activities

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The lipase activities in the pancreas and liver were analyzed according to the

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method described by Knarreborg et al., (2003) with slight modifications. The samples

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(1.0 g) were homogenized in 2 mL of phosphate buffered saline (PBS, pH = 7.4)

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using an Ultra Turrax (T25 basic IKA, Germany) in an ice bath before being

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centrifuged at 12000 g for 15 min at 4C. The supernatant was stored at -20C, and

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the lipase activity was analyzed within 24 h using ELISA lipase kit (CUSABIO

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Biotech Co., Ltd, Wuhan, China). Ten microliter of the supernatant was used when

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analyzing and comparing the lipase activities.

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2.6.3 Assays of lipogenic enzyme activities

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The lipogenic enzyme (FAS and ME) activities in the breast meat and liver were

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determined using a modification to the method described by Alvarez et al., (2000).

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Briefly, the samples (1.0 g) were homogenized in 3.0 mL of ice-cold extraction buffer

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(0.05 M Tris/HCl, pH 7.4; 0.5 M sucrose, 2 mM EDTA, 0.1 M NaF, 0.5 mM

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phenylmethylsulfonyl fluoride, 0.01 M mercaptoethanol, pH 7.4). After centrifugation

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at 12000 g for 30 min at 4 C, the supernatant was used to measure the enzymatic

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activities (Rosebrough et al., 2002). The ELISA kits (CUSABIO Biotech Co., Ltd,

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Wuhan, China) were used according to the manufacturer's instructions.

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The optical density (OD) was measured at 450 nm with a microtiter plate reader

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(Multiskan FC, Thermo Scientific, Vienna, Austria) to assay the AMPK, lipase and
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lipogenic enzyme activities.

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2.7 Statistical analysis

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For all analyses, surviving birds, pooled for each treatment served as the

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experimental replicates. The data were analyzed using SPSS statistical software

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(Version 19.0). All samples were used for experimental analysis, and single

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measurement was conducted per individual bird sample. An analysis of variance

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(one-way ANOVA by Duncan) was used to assess the differences between the

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treatments. A p-value of < 0.05 indicated significance. The data were expressed as the

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means standard deviations (SD).

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3. Results

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3.1 Effects of phosphorus level on growth performance and meat quality

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The effect of feeding diet with different P levels on feed consumption, feed

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conversion ratio (FCR) and growth performance of broiler chickens has been shown

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in Table 2. There were significant differences in final body weight among different

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dietary P levels (p < 0.001). The N group had the highest values. The daily weight

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gain of chickens fed with H1 and H2 diets was lower than N group (p < 0.05). The

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results indicated that the chickens reared on high P diets (H1 and H2) had lower feed

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consumption when compared to that fed the normal P diet (p < 0.05).

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The chemical composition of the breast meat from broiler chickens is presented in

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Table 3. Total phosphorus (TP) contents in breast meat reached 749 ~ 955 mg/kg,
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showing extremely significantly increase (p < 0.01) with increasing dietary P

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inclusion levels. No significant differences were observed in crude protein moisture

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and ash contents were observed in the treatment groups compared with that of N

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except H2 group. However, the moisture content of meat from poultry fed the highest

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P feed (H2) was significantly higher than the other three treatments (p < 0.01).

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The effects of the dietary P level on the cooking losses, shear forces, meat color and

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pH values are summarized in Table 3. The pH values and the cooking loss were not

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significantly different compared with the normal diet group. The shear force and meat

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color of breast meat showed extremely significant differences (p < 0.01) between

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different groups. The treatment groups exhibited higher lightness (L*), redness (a*)

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and shear force values than the N group (p < 0.05).

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3.2 Effects of phosphorus levels on lipid metabolism

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Table 3 shows that the serum concentrations of TC and TG were significantly

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higher in H1 and H2 groups than in N group (p < 0.05). The TG concentrations in the

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liver samples of H2 were significantly lower than in other groups (p < 0.05).

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The IMF content was an important indicator of meat quality. The IMF content of N

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group was 0.81 % higher than that of the treatment groups. The changes in the fatty

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acid composition of the breast meat are shown in Table 4. It can be seen that

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increasing dietary P levels decreased the fatty acid profiles significantly (p < 0.01)

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except for that of stearic acid (C18:0) (p = 0.16). Similarly, the SFA, MUFA, PUFA

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and total fatty acids decreased significantly with increasing of dietary P level (p <
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0.01).

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3.3 Effects of phosphorus level on enzyme activities

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The AMPK plays an important role during the development of fat cells and the

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deposition of IMF (Hardie, 2007). The AMPK activities were strongly influenced by

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the dietary P levels (Fig. 1). The AMPK activity in the breast meat and liver of N

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group is the lowest and that of D group is the highest. This result is contrary to the

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changes observed in the IMF content of the breast meat, which showed the IMF

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content of N group was significantly higher than that of treatment groups. The

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negative correlation between AMPK activity and IMF content observed in the present

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study is consistent with reported, which demonstrated that low AMPK activity is

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expected to enhance intramuscular fat (IMF) contents (Underwood et al., 2008).

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Therefore, the AMPK activity is probably negatively associated with the IMF content

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of breast meat.

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The changes in the lipase activity in the pancreas and liver in broiler chickens are

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shown in Fig. 1. Pancreatic lipase plays a substantial role during the digestion of

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lipids (Knarreborg et al., 2003). The activity of pancreatic lipase increased when the

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dietary P levels increased. The N group showed the highest lipase activity in liver

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with significant decreases (p < 0.01) as the dietary P level increased.

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The effect of the dietary P levels on the lipogenesis modulation capacity was

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assessed by determining the main lipogenic enzymes activities (ME and FAS) in

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broiler chickens (Fig. 2). It can be seen that ME activity in N group was the highest in
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the breast meat and the liver. Although the ME activities was lower in the

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experimental groups (D, H1 and H2) versus N group, the FAS activities increased

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continuously with the dietary P levels in breast meat. Whereas D group displayed

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higher activity than N group in the liver.

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4. Discussion

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The effects of the dietary P levels on the meat quality and lipid metabolism were

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investigated. The total P and nonphytate phosphorus requirements for broilers during

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the first three weeks and subsequent growth are 0.85 % and 0.80 %, 0.45 % and

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0.35 %, respectively (NRC, 1994). The P deficiencies or additions to feed in the

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broiler industry are common phenomena. Our results indicated that the high dietary P

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levels had a negative effect on growth performance of the chickens. The FI of

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chickens fed the high P diet was relatively lower than N group. TP content of the

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breast meat increased significantly when the dietary P levels increased (p < 0.001),

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indicating that excessive P consumption can lead to P deposits in muscle tissue. This

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is consistent with the results of Marounek et al., (2008), which also demonstrated that

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when the P intake in poultry diets reach a certain concentration, P levels in muscle

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improved as the P levels in the diet increase.

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The meat quality attributes including the meat color, pH value, cooking loss, and

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tenderness (Joo et al., 2013). Our results showed that the meat quality and chemical

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composition of breast meat were significantly affected when the dietary P inclusion

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levels increased. The chickens fed on diet excessive in P content (H2 group) had
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significant higher moisture and ash contents (p < 0.05) than other groups. In this study,

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the broiler chickens that consumed the experimental diets displayed significant impact

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on meat color and shear forces in breast meat. The shear force is an objective

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measurement of the tenderness of the meat (Lonergan et al., 2003). A lower shear

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force value indicates that the meat is tenderer. In the current study, the shear force

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values of the breast meat increased with increasing of dietary P levels, and the shear

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force of N group is close to what was reported by Wattanachant et al. (2004). Our

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results demonstrated that excessive or deficient P content in broiler diets adversely

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affect the meat color, moisture and tenderness of breast meat.

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In addition, the contents and composition of IMF are also important meat quality

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characteristics (Yang et al., 2009), and its content indirectly affects the sensory

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properties of meat, for example, tenderness and flavor, together with nutritional

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quality (Joo et al., 2013; Symeon, 2013). Our results showed that IMF contents in N

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group were significantly higher than that of deficient and high P group (p < 0.01),

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indicating IMF content can be affected by the dietary P levels. The fatty acid

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composition and biosynthesis play a role during the regulation for IMF contents

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(Camerona et al., 2000) and its profiles affect the nutritional and sensory

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characteristics of broiler chicken meat. Interestingly, breast meat from chickens fed

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with deficient P in diet had relatively high fatty acid content than other group, and

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high P dietary are likely to influence the SFA and PUFA, primarily the long chain

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fatty acids in broiler meat.


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The distribution and deposition of the IMF is affected by the activities of various

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key enzymes including ME, FAS and AMPK, which are related to lipid metabolism.

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Our results showed that ME activities in liver were lower in treatments than that of N

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group (p< 0.05), indicating the ME activity was inhibited when the P levels were high,

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and the degree of inhibition interacted with the IMF contents. Previous study showed

318

the changes in the activities of the lipogenic enzymes (FAS) can alter the rates of the

319

biosynthesis and hydrolysis of fatty acids (Smith, Witkowski, & Joshi, 2003).

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However, our results showed that the FAS activity in breast meat increased

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significantly with increasing of dietary P levels, and the FAS activity in the liver and

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breast meat displayed opposite trends relative to their fatty acid compositions.

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Therefore, lower body weight and decrease in the fatty acid levels observed in the

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treatment groups (D, H1 and H2) might be partly attributed to the decreased energy

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intake as a result of reduced feed consumption, and resulted fatty acid oxidation to

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meet the energetic demands than the P deficient group.

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Besides, our results showed that the IMF content decreased when the AMPK

328

activity increased in the breast meat, which agree with reported (Hardie, 2007;

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Underwood et al., 2008). This can be partly explained as fatty acid synthesis is one of

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the pathways most sensitive to AMPK inhibition (Underwood et al., 2008; Hardie et

331

al., 2003), and the activated AMPK can inhibit lipogenesis through phosphorylation,

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thus reduces the deposition of IMF (Hardie, 2007).

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The hydrolysis of TG mainly depends on lipase activity. Pancreatic lipase is a key


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enzyme during the absorption of dietary TG (Yang et al., 2014). Our results showed

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lipase activity significantly increased with increasing of dietary P level. Chickens fed

336

on diet adequate in P content had higher serum TC and TG concentrations than

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chickens fed on diet deficient in P content, because TG is broken down into blood to

338

supply energy for life activities, causing increasing serum TC and TG concentrations

339

in the exogenous lipid metabolism. Similarly, the TG concentration in the liver of H2

340

group is significantly lower than that of the other groups (p < 0.01). It is reported that

341

lipogenesis, the export of lipids, and synthesis and oxidation of fatty acid are

342

primarily achieved in the liver in endogenous lipid metabolism (Griffin et al., 1992;

343

Kersten, 2011), which are crucial steps for controlling TG accumulation in liver (Yang

344

et al., 2010). Previous reports have indicated that improving the serum lipid profile,

345

specifically increasing TC and TG, and decreasing the liver lipid accumulation and

346

lipid absorption are the most effective at decreasing the liver disorders (Yang et al.,

347

2014). During our study, high dietary P levels were not observed inducing fat

348

deposition in the liver. The results of lipid metabolism affected by P content in diet in

349

broilers may be explained from two aspects. On one hand, the metabolism of

350

lipoproteins may be facilitated. On the other, higher AMPK activity may inhibit the

351

TG synthesis in the liver. The effects of dietary P level on those parameters related to

352

lipid metabolism as supplementary materials, shown in Figure S1.

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5. Conclusion

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The present study investigated the effects of dietary P on meat quality and
17

355

indicators for lipid metabolism. Results showed that higher P level in diet for broiler

356

chicken resulted in higher P content in chicken breast meat. Deficient or high P

357

inclusion levels in broiler diets may negatively affect meat quality, resulting in

358

decrease on IMF contents and fatty acid of breast meat, increase shear force (less

359

tenderness) when compared with the normal group. Different P level in diet affect key

360

enzymes related to lipid metabolism. Deficient or high P groups had increased AMPK

361

activities in breast meat and the liver, FAS activities in liver and lowed lipase activity

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in liver, ME activities in breast meat and the liver compared with normal P group.

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Lipase activity in pancreas and FAS activity in breast meat increased with increasing

364

of dietary P level. Therefore, further studies on improving the quality of broiler

365

chicken meat by regulating lipid metabolism through fed diets with different P levels

366

are needed.

367

Acknowledgements

368

The authors would like to thank the Fund of Agro-scientific Research in the Public

369

Interest, China (Grant No. 201303083).

370

References

371

Alvarez, M. J., Diez A., Lopez-Bote C., Gallego M., & Bautista, J. M. (2000).

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Short-term modulation of lipogenesis by macronutrients in rainbow trout

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(Oncorhynchus mykiss) hepatocytes. British Journal of Nutrition, 84, 619-628.

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23

477

Figure captions

478

Figure 1. Effect of dietary phosphorus level on AMP-activated protein kinase (AMPK)

479

activities in breast meat (a) and liver (b) of broiler chickens. Effect of dietary

480

phosphorus level on lipase activities in pancreas (c) and liver (d) of broiler chickens.

481

The data are given as means SD. Different letters on data bars within each tested

482

parameter indicate a significant difference (p < 0.05). D, P deficient group; N, P

483

normal group; H1, high P group with available P 0.86% and 0.82%; H2, high P group

484

with available P 1.16% and 1.10%.

485

Figure 2. Effect of dietary phosphorus level on malic enzyme (ME) activities in

486

breast meat (a) and liver (b) of broiler chickens. Effect of dietary phosphorus level on

487

fatty acid synthase (FAS) activities in breast meat (c) and liver (d) of broiler chickens.

488

The data are given as means SD. Different letters on data bars within each tested

489

parameter indicate a significant difference (p < 0.05). D, P deficient group; N, P

490

normal group; H1, high P group with available P 0.86% and 0.82%; H2, high P group

491

with available P 1.16% and 1.10%.

24

24

AMPK activity (U/L)

26

AMPK in meat

36

22
20

18

ab

16
14

28

24

c
20
16
12

12

H1

H2

80

lipase in pancreas

76

H1

H2

Treatment

Treatment

AMPK in liver

32

AMPK activity (U/L)

36

ab

lipase in liver

72

68
64

lipase activity (U/L)

lipase activity (U/L)

34

60
56

a
32

30

28

52
26

492
493

H1

H2

H1

Treatment

Treatment

Figure 1.

494

25

H2

36

b 44

ME in meat

ME in liver
40

ME activity (U/L)

ME activity (U/L)

32
28

24
20

16

36
32
28

24

12

20

H1

H2

FAS in meat

FAS activity (U/L)

90
80

60

a
a

40

78
72

66

54

496

84

60

30

495

FAS in liver

90

70

50

H2

d 96

100

FAS activity (U/L)

H1

Treatment

Treatment

H1

H2

H1

Treatment

Treatment

Figure 2.

26

H2

Table 1: The composition and nutrient levels of diets for broiler chickens (air dry basis)
Starter phase (1~21 d)
D

Grower phase (22~42 d)

H1

H2

H1

H2

Ingredients, %
Corn
Soybean meal
Rapeseed meal
Cottonseed meal
Fish meal
Limestone
CaHPO4
KH2 PO4
NaCl
Premix2
Total

61.40
19.50
6.00
5.00
3.00
1.50
0.10
0.20
0.30
3.00
100

60.00
20.50
5.50
5.00
3.00
1.10
0.60
1.00
0.30
3.00
100

60.70
18.70
3.80
4.00
6.00
0.80
0.70
2.00
0.30
3.00
100

59.00
19.00
4.50
3.55
5.60
0.85
0.70
3.50
0.30
3.00
100

62.00
19.00
4.70
5.00
4.50
1.50
0
0
0.30
3.00
100

62.00
19.00
4.00
3.50
5.90
1.20
0.20
0.90
0.30
3.00
100

61.00
18.50
3.80
4.00
6.00
0.90
0.50
2.00
0.30
3.00
100

59.70
19.10
4.10
3.50
5.60
0.80
0.70
3.20
0.30
3.00
100

Nutrient levels3
ME/(MJ/kg)
CP %
Ca %
TP %
AP %
Lys %
Met %
Met + Cys %

12.55
20.20
0.90
0.55
0.28
1.11
0.34
0.68

12.45
20.31
0.90
0.85
0.56
1.12
0.34
0.68

12.45
20.28
0.92
1.15
0.86
1.16
0.36
0.68

12.22
20.08
0.92
1.47
1.16
1.15
0.36
0.68

12.65
20.42
0.91
0.51
0.26
1.15
0.35
0.69

12.64
20.30
0.91
0.78
0.52
1.17
0.36
0.69

12.47
20.22
0.90
1.10
0.82
1.17
0.36
0.69

12.30
20.01
0.91
1.40
1.10
1.16
0.35
0.68

1) D, P deficient group; N, P normal group;H1, high P group with available P 0.86% and 0.82%; H2, high P group with available P 1.16% and 1.10%. 2) The premix
provided the following per kg of diets: VA 15000 IU, VD3 2700 IU, VE 60 mg, VB1 3 mg, VB2 9 mg, VB6 3 mg, VB12 0.03 mg, nicotinic acid 60 mg, pantothenic
acid 15 mg, biotin 0.36 mg, choline chloride 600 mg, Fe 90 mg, Cu 12 mg, Zn 75 mg, Mn 90 mg, I 0.35 mg, Se 0.25 mg. 3) Ca and TP were measured values, while
27

the others were calculated values.

28

Table 2: Effect of dietary phosphorus level on growth performance in broiler chickens


Treatments
Items

p-value

H1

H2

Initial/g

41.523.65

41.892.39

42.692.08

42.143.30

0.637

Final/g

1411135.56b

159796.68a

108094.54c

88370.27d

<0.001

BWG/g

32.543.99a

37.235.10a

24.584.63b

20.143.11b

0.002

FI (g/bird per d)

56.017.28ab

64.2312.00a

45.476.61bc

39.884.21c

0.012

FCR (g/g)

1.740.13b

1.720.10b

1.850.06ab

1.980.08a

0.022

Mortality/%

12

12

16

Weights

D, P deficient group; N, P normal group; H1, high P group with available P 0.86% and 0.82%; H2, high
P group with available P 1.16% and 1.10%. BWG, body weight gain; FI, feed intake; FCR, feed
conversion ratio.

29

Table 3: Effect of dietary P level on chemical composition and lipid metabolism in broiler chickens
Items
Moisture/ %
Crude
protein/ %
Ash/ %
Total
phosphorus
(mg/kg)
Shear force
(N)
Cooking
loss (%)
L*
a*
b*
pH value
lipid
metabolism
Serum TC
(mol/L)
Serum TG
(mmol/L)
TG in Liver
(mmol/L)
IMF in
muscle/ %

Treatments

p-value

H1

H2

73.590.30a

73.740.36a

73.770.39a

75.130.37b

<0.001

24.40.95

24.150.44

23.840.79

23.451.23

0.457

1.590.20ab

1.710.13a

1.680.14a

1.340.19b

0.037

74967a

87637b

90735bc

95523c

<0.001

11.430.70b

8.950.72c

10.591.02b

12.920.64a

<0.001

22.950.74

23.191.06

23.510.60

24.531.87

0.312

53.562.21ab
1.410.48ab
4.190.85a
5.730.13

51.871.72a
1.040.29a
5.390.96b
5.840.12

52.621.38a
1.860.51b
4.770.99ab
5.760.13

55.311.55b
2.430.17c
5.801.31b
5.730.08

0.009
0.001
0.032
0.205

3.390.31ab

3.200.15a

3.560.33b

3.680.34b

0.041

0.370.04ab

0.340.03a

0.440.06bc

0.460.07c

0.009

3.390.29a

3.560.05a

3.480.14a

2.550.39b

0.004

0.620.02a

0.810.03c

0.700.01b

0.610.01a

<0.001

D, P deficient group; N, P normal group; H1, high P group with available P 0.86% and 0.82%; H2, high
P group with available P 1.16% and 1.10%. Results are represented as mean SD. Dierent letters in
the same row indicate significant dierences (p < 0.05). L*, Lightness; a*, Redness; b*, Yellowness.
TC= Total cholesterol; TG= Triglyceride; IMF= Intramuscular fat contents.

30

Table 4: Effect of dietary P level on fatty acids profile of breast meat in broiler chickens
Treatments
Items

p-value

H1

H2

C16:0

141.659.22a

123.994.36b

123.454.53b

95.07.35c

<0.001

C16:1

15.410.17a

13.941.43a

10.580.04b

7.481.21c

<0.001

C18:0

83.492.14

76.423.97

75.622.71

74.967.48

0.160

C18:1

121.653.31a

92.548.94c

104.024.86b

55.912.26d

<0.001

C18:2

132.888.09a

108.935.84c

120.335.98b

91.33.29d

<0.001

C18:3

5.00.21a

3.950.56bc

4.430.45ab

3.190.34c

0.004

SFA

225.1411.36a

200.418.33b

199.077.24b

169.9614.83c

0.002

MUFA

137.063.48a

121.8510.37b

114.64.9b

63.393.47c

<0.001

PUFA

137.888.3a

112.229.81c

124.768.69b

94.483.63d

<0.001

Total fatty
acids

500.0823.14a

419.1125.97b

438.435.31b

327.8319.35c

<0.001

D, P deficient group; N, P normal group; H1, high P group with available P 0.86% and 0.82%; H2,
high P group with available P 1.16% and 1.10%. Results are represented as mean SD and
expressed as mg FA/100 g fresh meat. Dierent letters in the same row indicate significant
dierences (p < 0.05). SFA= saturated fatty acid; MUFA= mono-unsaturated fatty acid; PUFA=
poly-unsaturated fatty acid.

31

1. Meat quality and enzyme activities were determined in chickens.


2. Dietary P levels influenced meat quality and expression of indicators on lipid
metabolism of broilers.
3. Meat quality of broiler chickens is decreased compared to normal group.
4. Provide useful information for improvement of the meat quality.

32

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