1. Introduction
The consumption of poultry meat products has continued to increase due to their low price, high nutritional value, absence of a religious effect, and suitability for processing [
1]. Throughout the poultry industry, there have been tremendous improvements in growth rates and breast meat yield, resulting in substantial increases in commercial meat production. Nonetheless, these advances are associated with several significant implications in the quality of the meat obtained [
2]. The white striping (WS) condition present in broiler breast fillets is characterized by the appearance of white striations running parallel to the muscle fibers on the surface of the pectoralis major muscle [
3]. For consumers, visual appearance is the primary and most important attribute to assess the quality of a meat product in a sealed package. Thus, the presence of any condition negatively affecting the visual appearance of a product can influence the purchase decision, leading to potential economic loss [
3]. In addition to product appearance, the presence of white striping can become a hindrance, in regard to further processing, leading to a reduction in the water holding capacity of a meat product during processing or storage [
4], or poor product cohesion related to immaturity of intramuscular connective tissue [
5].
Skeletal muscle mass and muscle fiber vary in size according to physiological and pathological conditions. An increase in muscle mass, muscle growth or hypertrophy occurs during development and in response to mechanical overload or exercise [
6]. Mechanical stimuli play a significant role in the regulation of skeletal muscle mass [
7]. The mammalian target of rapamycin (mTOR), a protein kinase, determines the mechanical regulation of muscle mass [
7]. It has been shown that the activation of mTOR signaling is sufficient to induce an increase in muscle fiber size [
7]. The mechanical activation of mTOR signaling involves phospholipase D (PLD) and the lipid second messenger phosphatidic acid (PA). Phosphatidic acid (PA) is a diacyl-glycerophospholipid, in which two fatty acids and a phosphate group are covalently bonded to a glycerol molecule through ester linkages. PA can act as a signaling lipid, is a precursor for the biosynthesis of other lipids, and is a major constituent of cell membranes. Intracellular concentrations of PA can increase as a result of mechanical stimuli. During mechanical stimulation, PA can activate mTOR signaling via a P13K- and ERK-independent mechanism, bind to the FKBP12-rapamycin binding (FRB) domain of mTOR, and directly activate mTOR kinase activity in vitro [
7]. Hence, a mechanically induced increase in PA could lead to enhanced binding of PA and mTOR, resulting in the activation of mTOR signaling and ultimately muscle growth [
7]. It has been demonstrated that exogenous PA can stimulate the mTOR pathway via its activation of the substrate S6 kinase [
8,
9]. However, the binding of PA to S6 kinase may occur independently of mTOR [
10], suggesting that PA may augment the signaling response when mTOR is activated by exercise.
While PA can be synthesized from a variety of reactions via multiple reactants, it is not clear if other precursors (i.e., glycerol-3-phosphate (G3P), LPA, or diacylglycerol (DAG)), or the addition of head groups to the PA molecule (i.e., phosphatidylcholine (PC), phosphatidylserine (PS), phosphatidylethanolamine (PE), or phosphatidylinositol (PI)), have a similar ability to activate mTOR signaling. Furthermore, different sources of PA (e.g., soy, egg) can have varying degrees of unsaturated or saturated fatty acid chains which can influence the behavior of PA. Foster [
11] suggested that two saturated fatty acids will promote storage, but one saturated and one unsaturated fatty acid will promote signaling. In addition to controlling the rate of protein synthesis, mTOR also regulates transcriptional changes in response to a variety of conditions, such as cell cycle progression, actin organization, autophagy, synaptic plasticity, memory, and learning [
12]. In turn, both intrinsic and extrinsic factors may have a direct impact on the mTOR pathway, due to the number of processes modulated by mTOR [
12]. The mTOR signaling network is wired to growth factor signaling via the insulin/insulin-like signaling system (IIS) [
13]. Decreased IIS/TOR signaling activity has been associated with an increased resistance to certain types of stress, indicating the pathway plays a vital role in the adaption to different stress conditions [
14,
15]. The IIS/mTOR signaling is positively correlated with muscle growth. Therefore, IIS and mTOR pathways play an important role in hypertrophic muscle accumulation [
16].
The objective of this study was to evaluate the effect of dietary PA supplementation on broiler chicken growth and meat yield. To accomplish this, two experiments were conducted. The first examined if exercise was needed along with PA supplementation to increase muscle growth in broiler chickens. The second experiment evaluated different inclusion rates of PA in the feed. It was hypothesized that PA supplementation will increase muscle growth and yield, and that exercise will increase each of these factors.
4. Discussion
The hypothesis that feeding PA along with exercise would result in increased growth specifically in the breast muscle was rejected. The addition of exercise did not increase the growth or meat yield in Experiment 1. However, increases were observed in birds receiving dietary PA. Experiment 2 also observed an increase in growth and meat yield at the low inclusion rate. This does not agree with the research in humans [
17,
18,
19], which observed that exercise in conjunction with dietary PA increased muscle mass. In the human studies it is thought that PA and exercise work together to increase muscle growth, as PA is a signaling lipid in the mTOR pathway, and is increased in mechanical stimulation during exercise [
7]. However, it has also been demonstrated that exogenous supplied PA can result in stimulation of the mTOR pathway without exercise [
8,
9] so it is possible that exercise is not needed in poultry to achieve increased muscle growth.
In addition, the method used to exercise birds during this study likely resulted in stress. Negative attributes associated with stress, such as decreased growth and feed conversion have been well documented [
20], thus, counteracting any positive effects resulting from the combination of exercise and PA. This is supported by the results observed in Experiment 2 which indicate increases in growth and meat yields with solely dietary inclusion of PA without exercise
No differences were observed in the woody breast or white striping scores in Experiment 2. This is important, as the severity of white striping and woody breast have increased in recent years. White striations characterize white striping parallel to muscle fibers in which muscle fiber degeneration takes place with infiltration of fat and connective tissue [
21,
22,
23]. Woody breast is characterized by pale broad areas of substantial hardness accompanied with white striations [
24]. Woody breast also results in hard, rigid fillets in which muscle fiber degeneration takes place with infiltration of connective tissue [
24,
25]. Sihvo [
25] concluded that fast growth rate, along with increased breast meat yield, plays a significant role in the development of woody breast in chicken. Higher growth rates are also associated with greater incidence of white striping in poultry [
21]. With evidence indicating that both woody breast and white striping are a result of fast growth and muscle degeneration, it is possible that utilizing PA to stimulate the mTOR pathway could be the reason this study observed increased growth and meat yield. It is possible that stimulating muscle growth via the mTOR pathway [
7] somehow counteracts or prevents the muscle degeneration observed in white striping and woody breast, although this requires further research to confirm.
In Experiment 2, the high inclusion rate did not see the same increased growth that it did in Experiment 1. It is possible that other environmental factors may affect high levels of dietary PA to increase growth in poultry. An example is stress. Stress affects the mTOR signaling pathway [
13]. It is possible that there were external stressor differences, such as temperature that altered the effect of PA at different inclusion rates. This, however, merits further research to determine if this hypothesis is true. The mTOR pathway is also involved in immune function [
26] and gut health [
27] which are two factors not measured in this study but merit investigation.