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<article article-type="research-article" dtd-version="1.1" xml:lang="en" xmlns:mml="http://www.w3.org/1998/Math/MathML" xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance">

	<front>
		<journal-meta>
			<journal-id journal-id-type="publisher-id">SJAR</journal-id>
			<journal-title-group>
				<journal-title>Spanish Journal of Agricultural Research</journal-title>
				<abbrev-journal-title>Span J Agric Res</abbrev-journal-title>
			</journal-title-group>
			<issn pub-type="epub">2171-9292</issn>
			<publisher>
				<publisher-name>Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA)</publisher-name>
			</publisher>
		</journal-meta>
		<article-meta>
			<article-id pub-id-type="publisher-id">19542</article-id>
			<article-id pub-id-type="doi">10.5424/sjar/2023211-19542</article-id>
			<article-categories>
				<subj-group subj-group-type="heading">
					<subject>RESEARCH ARTICLE</subject>
				</subj-group>
			</article-categories>

			<title-group>
				<article-title>Performance and meat quality characteristics of male quails (<em>Coturnix coturnix japonica</em>) fed diets supplemented with pomegranate seed oil</article-title>
			</title-group>

			<contrib-group>
				<contrib contrib-type="author">
					<contrib-id contrib-id-type="orcid">https://orcid.org/0000-0002-5507-5830</contrib-id>
					<name>
						<surname>Sarmiento-García</surname>
						<given-names>Ainhoa</given-names>
					</name>
					<aff id="aff1"><institution>Departmento de Construcción y Agronomía, Facultad de Ciencias Agrarias y Ambientales, Universidad de Salamanca,</institution><addr-line>37007 Salamanca,</addr-line><country> Spain.</country></aff>
				</contrib>
				<contrib contrib-type="author">
					<contrib-id contrib-id-type="orcid">https://orcid.org/0000-0003-2309-2473</contrib-id>
					<name>
						<surname>Gökmen</surname>
						<given-names>Seyit A.</given-names>
					</name>
					<aff id="aff2"><institution>Department of Animal Science, Faculty of Agriculture, Selcuk University, </institution><addr-line>42130 Selcuklu, Konya, </addr-line> <country>Türkiye.</country></aff>
				</contrib>
				<contrib contrib-type="author">
					<contrib-id contrib-id-type="orcid">https://orcid.org/0000-0003-2996-3241</contrib-id>
					<name>
						<surname>Sevim</surname>
						<given-names>Behlül</given-names>
					</name>
					<aff id="aff3"><institution>Eskil Vocational School, Aksaray University, </institution><addr-line>68800 Eskil, Aksaray, </addr-line><country>Türkiye.</country></aff>
				</contrib>
				<contrib contrib-type="author">
					<contrib-id contrib-id-type="orcid">https://orcid.org/0000-0002-3732-1137</contrib-id>
					<name>
						<surname>Olgun</surname>
						<given-names>Osman</given-names>
					</name>
					<aff id="aff2"><institution>Department of Animal Science, Faculty of Agriculture, Selcuk University, </institution><addr-line>42130 Selcuklu, Konya, </addr-line> <country>Türkiye.</country></aff>
				</contrib>				
							
			</contrib-group>
			<pub-date pub-type="epub">
				<day>27</day>
				<month>01</month>
				<year>2023</year>
			</pub-date>			
			<pub-date pub-type="collection">
				<month>03</month>
				<year>2023</year>
			</pub-date>
			<volume>21</volume>
			<issue>1</issue>
			<elocation-id>e0602</elocation-id>
			<history>
				<date date-type="received">
					<day>04</day>
					<month>05</month>
					<year>2022</year>
				</date>
				<date date-type="accepted">
					<day>27</day>
					<month>01</month>
					<year>2023</year>
				</date>
				<date date-type="pub">
					<day>27</day>
					<month>01</month>
					<year>2023</year>
				</date>
			</history>			
			<permissions>
				<copyright-statement>&#xa9;2023 CSIC</copyright-statement>
				<copyright-year>2023</copyright-year>
				<license license-type="open-access" xlink:href="https://creativecommons.org/licenses/by/4.0/">
					<license-p>This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International (CC BY 4.0) License.</license-p>
				</license>
			</permissions>
			<self-uri xlink:href="https://doi.org/10.5424/sjar/2022204-19567"/>
			<abstract>
				<title>Aim of study:</title>
				<p>Pomegranate seed oil (PSO) is a nutritive, antioxidant-rich by-product, and it has been tested as a feed ingredient for livestock. However, studies on quails are scarce. The current study investigated that the effect of PSO on the performance and meat instrumental quality of quails.</p>
				<title>Area of study:</title>
				<p>Türkiye.</p>
				<title>Material and methods:</title>
				<p>A total of 60 seventy-day-old male quails were equally subjected to 3 dietary treatments consisting of 20 birds (5 replicates with 4 birds each). The quails were fed a diet supplemented with 0, 100, and 200 mg kg<sup>-1</sup> pomegranate seed oil (PSO). After 10 weeks, two birds per subgroup were randomly selected and slaughtered.</p>
				<title>Main results:</title>
				<p>Supplementation of PSO reduced (p&lt;0.05) feed intake and body weight gain linearly. However, there was no significant effect of PSO on carcass traits. For color parameters, the L* and b* values of breast and thigh meat increased with the addition of PSO to the diet (p&lt;0.05). Cooking losses were highest in the thigh of quails fed 200 mg kg-1 PSO. On the other hand, in the breast, the lowest values for this parameter were observed in the groups that had received 100 mg kg<sup>-1</sup> of PSO.</p>
				<title>Research highlights:</title>
				<p>Including 100 mg kg<sup>-1</sup> of PSO can improve some meat quality characteristics without affecting performance parameters. There is a possibility that meat quality could be negatively affected by values higher than this. Nevertheless, further research is needed to determine the optimal dose of PSO to improve quail meat quality and its performance.</p>									
			</abstract>
			<kwd-group>
				<kwd>meat and carcass quality;</kwd>
				<kwd>birds;</kwd>
				<kwd>quail performances;</kwd>
				<kwd>pomegranate oil.</kwd>
			</kwd-group>
		</article-meta>
		<funding-group id="fw-01">
			<award-group id="aw1">
				<funding-source>The authors received no specific funding for this work</funding-source>
			</award-group>
		</funding-group>
		<p><strong>Ethics approval statement:</strong> The authors confirm that the ethical policies of the journal, as noted on the journal’s author guidelines page, have been adhered to and the appropriate ethical review committee approval has been received. The European National Research Council's guidelines for the Care and Use of Laboratory Animals were followed.</p>		
	</front>
	<body>
		<sec id="sec1" sec-type="intro">
			<title>Introduction</title>

			<p>Pomegranate (<em>Punica granatum</em> L.), from the family Punicaceae, is one of the oldest edible fruits and is commonly cultivated in many tropical and subtropical countries (Boroushaki et al., 2016; Białek et al., 2021a). Pomegranate fruits are mainly consumed fresh or processed, however, significant amounts of pomaces are generated during processing, which causes serious environmental hazards (Białek et al., 2021a). Recently, the utilization of these wastes as a source of valuable compounds which could be used as an ingredient in animal nutrition has received particular attention (Saki et al., 2014; Boroushaki et al., 2016; Białek et al., 2019, 2021a).</p>

			<p>It is estimated that pomegranate seeds contain approximately 24% oil (Khoddami et al., 2014) and the fatty acids present in them are of particular interest (Białek et al., 2021a). Punicic acid (9‐cis, 11‐trans, 13‐cis or trichosanic acid) is the dominant form of the C18:3 class, which has a positive effect on health due to its therapeutical properties (Khoddami et al., 2014; Boroushaki et al., 2016). Studies have shown that pomegranate seed oil (PSO) can affect multiple signaling pathways involved in inflammation, cellular transformation, hyperproliferation, angiogenesis, initiation of tumorigenesis, and eventually suppressing the final steps of tumorigenesis and metastasis (Sharma et al., 2017). For example, Białek et al. (2021b) and Teh et al. (2019) demonstrated that PSO could improve cardiovascular health by reducing total cholesterol. Likewise, Mehta &amp; Lansky (2004) described that PSO could be a potential breast cancer prevention, and Lansky et al. (2005) proposed that PSO could be a potent inhibitor of human PC-3 prostate cancer cells.</p>

			<p>Japanese quails are valuable animals for avian research. Their small size makes them easy to handle, which allows them to be stored in limited spaces (Minvielle, 2004; Alkan et al., 2010). Moreover, farmer interest in raising quail for marketing continues to grow in Japan, France, Indonesia, Spain and Brazil (Minvielle, 2004; Santos et al., 2011). Japanese quails are commonly raised not only to produce eggs but also to produce meat in some countries. It is expected that in the near future, quail will become the main poultry used for meat production, due to its widespread demand around the world (Sabow, 2020). Thus, researchers should focus on improving the meat quality and productive performance of these birds.</p>

			<p>Changes in animals’ feeding are the most commonly studied and applied way to modify meat properties and quality (Sarmiento-García et al., 2021; Vieira et al., 2021). It is also an indirect method of improving the diet, which in turn will have a health-promoting effect, and the appearance of the product (Saki et al., 2014). PSO also has significant effects on meat quality and the development of livestock species (Saki et al., 2014; Karampour &amp; Kafilzadeh, 2016; Szymczyk &amp; Szczurek, 2016; Emami et al., 2017; Białek et al., 2021b) no study has been conducted on the effects of pomegranate as a feed additive on the yield and instrumental meat quality of quails. The aim of this study was to evaluate the effects of dietary PSO on performance, carcass traits, as well as the instrumental quality of quail’s thigh and breast meat.</p>

		</sec> <!--sec1-->

		<sec id="sec2" sec-type="materials|methods">
			<title>Material and methods</title>
			
			<sec id="sec2.1">
				<title>Ethical approval</title>

				<p>Criteria specified by European policy for protecting animals were followed during the experimental period (EU, 2010).</p>

			</sec> <!--sec2.1-->

			<sec id="sec2.2">
				<title>Birds, feeding, and management</title>

				<p>The whole experiment was conducted on 60 male Japanese quails (<em>Coturnix coturnix Japonica</em>) for 10 weeks at Selçuklu, Konya, Türkiye (38°1’36, 32°30’45”). Quails with similar body weight (180.65±5.30 g) and 70 days of age were obtained from a commercial company. The trial was conducted in 3 experimental groups consisting of 5 replicates, each containing 4 male quails.</p>

				<p>Animals were allocated randomly to different cages (30 × 45 cm) that had the same environmental conditions. Birds were housed in a well-ventilated room with a lighting program of 16 hours, and a temperature of 20±2.0 °C was maintained in each pen.</p>

				<p>Three dietary treatments were tested consisting of three graded levels (0, 100, and 200 mg kg<sup>-1</sup>). All iso-nitrogenous (24% crude protein) and iso-energetic (2900 kcal/kg metabolizable energy) diets were formulated according to the NRC (1994). During the experimental period, all quails received feed and water <em>ad-libitum</em>. PSO included in the diet was commercially available and purchased at a local market (Arı Mühendislik, Ankara, Türkiye). It was stored at 4°C prior to the preparation of the experimental diets and administration to quails. A basal diet was prepared without soybean oil, and then the soybean oil and PSO required for each treatment were mixed and added to the diet. The chemical composition of the basal diet was analysed according to AOAC (2006) procedures: water content by drying at 105 ºC; protein content by the Kjeldahl method; fat content by Soxhlet extraction; and ash content by incineration. The ingredients and nutrient composition of the basal diet are shown in Table 1.</p>

			</sec> <!--sec2.2-->

			<table-wrap id="t1">
				<label>Table 1</label>
					<caption>
						<title>Ingredients and nutrient composition of basal diet.</title>
					</caption>
					<table>
						<thead>
							<tr>
								<th align="center">Ingredients</th>
								<th align="center">%</th>
								<th align="center">Nutrient contents</th>
								<th align="center"></th>
							</tr>							
						</thead>
						<tbody>
							<tr>
								<td>Corn</td>
								<td align="center">53.70</td>
								<td>Metabolizable energy (kcal ME/kg)</td>
								<td align="center">2899</td>
							</tr>
							<tr>
								<td>Soybean meal</td>
								<td align="center">40.00</td>
								<td>Dry matter (%)</td>
								<td align="center">87.32</td>
							</tr>
							<tr>
								<td>Soybean oil</td>
								<td align="center">2.50</td>
								<td>Crude protein (%)</td>
								<td align="center">24.03</td>
							</tr>
							<tr>
								<td>Limestone</td>
								<td align="center">1.06</td>
								<td>Ether extract (%)</td>
								<td align="center">5.53</td>
							</tr>
							<tr>
								<td>Dicalcium phosphate</td>
								<td align="center">1.90</td>
								<td>Crude fiber (%)</td>
								<td align="center">3.70</td>
							</tr>
							<tr>
								<td>Salt</td>
								<td align="center">0.35</td>
								<td>Ash (%)</td>
								<td align="center">5.97</td>
							</tr>
							<tr>
								<td>Premix <sup>[1]</sup></td>
								<td align="center">0.25</td>
								<td>Calcium (%)</td>
								<td align="center">1.00</td>
							</tr>
							<tr>
								<td>DL methionine</td>
								<td align="center">0.24</td>
								<td>Available phosphorus (%)</td>
								<td align="center">0.50</td>
							</tr>
							<tr>
								<td>Total</td>
								<td align="center">100.00</td>
								<td>Lysine (%)</td>
								<td align="center">1.31</td>
							</tr>
							<tr>
								<td></td>
								<td align="center"></td>
								<td>Methionine (%)</td>
								<td align="center">0.52</td>
							</tr>
							<tr>
								<td></td>
								<td align="center"></td>
								<td>Cystine (%)</td>
								<td align="center">0.44</td>
							</tr>
							<tr>
								<td></td>
								<td align="center"></td>
								<td>Methionine + cystine (%)</td>
								<td align="center">0.96</td>
							</tr>
						</tbody>
					</table>
					<table-wrap-foot>
						<fn id="TFN1">
							<p><sup>[1]</sup> Vitamin-mineral premix supplied per kilogram of diet: vitamin A (trans-retinol), 3.60 mg; vitamin D3 (cholecalciferol), 0.10 mg; vitamin E (α-tocopherol acetate), 75.00 mg; vitamin K3 (menadione), 5.00 mg; vitamin B1 (thiamine), 3.00 mg; vitamin B2 (riboflavin), 6.00 mg; vitamin B6 (pyridoxine), 5.00 mg; vitamin B12 (cyanocobalamin), 0.03 mg; nicotinic acid, 40.00 mg; pantothenic acid, 10.00 mg; folic acid, 0.75 mg; D-biotin, 0.00 mg; choline chloride, 375.00 mg; manganese (manganese oxide), 80.00 mg; iron (ferrous carbonate), 40.00 mg; zinc (zinc oxide), 60.00 mg; copper (cupric sulphate pentahydrate), 5.00 mg; iodine, 0.15 mg; selenium (sodium selenite), 0.30 mg.</p>
						</fn>
					</table-wrap-foot>
				</table-wrap>




			<sec id="sec2.3">
				<title>Determination of performance parameters</title>

				<p>At the beginning of the experiment, the quails were randomly allocated to the three trial groups. Animals were weighed individually at the beginning and the end of the experiment with a precision weighing scale (±0.01 g). Experimental diets were given by weighing each subgroup, and subsequently, feed intake (FI) was calculated as the daily FI per quail (g). Then, average body weight gain (g/day) was calculated by subtracting initial body weight (g) from final body weight (g) over the study period.</p>

			</sec> <!--sec2.3-->

			<sec id="sec2.4">
				<title>Meat characteristics analysis</title>

				<p>At the end of the experiment, all quails were weighed, and 30 quails (two birds from each subgroup) were randomly selected, slaughtered and then, the carcass yield was obtained by separating the head, feet, feathers, digestive organs and giblets. After 15 min post-mortem, abdominal fat, breast, and leg muscles, liver, pancreas, and heart were excised and weighed. Carcass yield was calculated as the ratio of the eviscerated carcass mass to body weight. The breast (<em>Pectoralis major</em>) with keel bone and left thigh muscle and drumstick were weighed and carried out to the laboratory (Faculty of Agriculture, Selcuk University) for analysis. To determine the meat yield, breast, thigh and drumstick were calculated as a percentage of carcass weight. Carcass yield, abdominal fat, liver, heart and pancreas were calculated as a percentage of body weight.</p>

				<p>Prior to the analysis, the skin of the breast and thigh were removed. Instrumental quality parameters on breast and thigh muscles were carried out according to Sarmiento-García et al. (2021) except for drip losses. All measurements were analysed in triplicate to minimize sampling error.</p>

			</sec> <!--sec2.4-->

			<sec id="sec2.5">
				<title>pH of muscles</title>

				<p>Muscle pH was determined at 24 hours post-mortem with a Crison pH Meter Basic 20® (Hach Lange Spain, L’Hospitalet de Llobregat, Barcelona, Spain) equipped with a penetration glass electrode. It was inserted directly in the thickest part of both muscles. The pH meter was calibrated using the 2-point method against standard buffer solutions with pH values of 4.0 and 7.0.</p>

			</sec> <!--sec2.5-->

			<sec id="sec2.6">
				<title>Colour</title>

				<p>For instrumental colour measurement, a colourimeter HunterLab MiniScan model XE Plus (Hunterlab, VA, USA) equipped with a 25 mm measuring head and diffuse/8° optical geometry was used. The meat colour was measured on the dorsal side of the breast and the medial (bone) side of the thigh, at 24 hours post-mortem, determining the parameters L* (lightness), a* (redness), and b* (yellowness) using a MiniScan XE Plus in the CIE L* a* b* space under D65, 10°, and Specular Component Included (SCI) conditions.</p>

			</sec> <!--sec2.6-->

			<sec id="sec2.7">
				<title>Cooking losses</title>

				<p>To determine cooking losses, the muscles were stored at 4°C until 24 h post-mortem, weighed and packaged in plastic bags, and heated to 75°C (centcenterpiecer 30 min, and then cooled in cold running tap water for 20 min. The internal temperature of the muscles was measured with a ChecktemW1 digital thermometer (Hanna Instruments, Eibar, Spain). The mass changes were expressed as a percentage of the initial mass.</p>

			</sec> <!--sec2.7-->

			<sec id="sec2.8">
				<title>Drip losses</title>

				<p>The drip losses of meat were estimated according to a modification of the method proposed by Zhang et al. (2015). Briefly, meat samples with a size of 3 cm (length) × 2 cm (width) × 1 cm (thickness), were weighed (W1) and suspended parallel to the longitudinal axis of the myofibers in netting and vacuumed bags and stored at 4°C. Samples were weighed after 24 h (W2). The difference in weight (W2-W1) corresponded to the drip loss and was expressed as the percentage of the initial muscle weight.</p>

			</sec> <!--sec2.8-->

			<sec id="sec2.9">
				<title>Statistical analysis</title>

				<p>A one-way ANOVA was used to test the effect of the experimental diets on performance, carcass traits, and instrumental meat quality in the quails. If ANOVA showed significant differences among means (main effects), planned multiple comparisons of means were examined by Duncan’s multiple range test. The statistical differences were defined as p &lt; 0.05 and trends as p &lt; 0.10. Orthogonal polynomial contrasts were used to assess the significance of linear and quadratic models to describe the response of the dependent variable to rising dietary PSO levels. All statistical analyses were carried out using the SPSS Package 23 (IBM SPSS Statistic, 2017).</p>

			</sec> <!--sec2.9-->

		</sec> <!--sec2-->

		<sec id="sec3" sec-type="results">
			<title>Results</title>

			<p>No mortality or illness symptoms were observed during the entire trial, and all quails were alive at the end of the experiment. According to the data in Table 2, the absence of statistical differences between the initial weights of the quails indicates that the subjects were randomly assigned to the experimental groups. PSO-supplemented diets linearly affected quail body weight gain and FI (p&lt;0.05). These parameters decreased linearly with the addition of PSO to the diet. Likewise, a trend (p=0.092) was observed for final weight, which decreased as PSO concentration in the diet increased. However, a linear effect of PSO inclusion (p &lt;0.05) on final weight was observed.</p>

			<p>The effects of the dietary supplementation of PSO on the carcass traits are given in Table 3. Although there was a trend in final body weight between the experimental groups, no statistically significant differences in carcass traits were found between the groups (p&gt;0.05). The minimum and maximum values for those parameters were as follows: carcass yield (59.39–60.38 %), abdominal fat (1.21–1.72 %), breast (53.04–54.28 %), thigh and drumstick (34.86–36.63 %), liver (1.33–1.43 %), heart (0.79–0.82 %), and pancreas (0.17–0.18 %).</p>

			<p>The meat colour was expressed as lightness (L∗), redness (a∗), and yellowness (b∗) values (Table 4). Breast meat from quail supplemented with PSO was paler and yellower than breast meat samples collected from control quails (p&lt;0.001). Similar results were observed for the thigh meat, where the highest L* value (p&lt;0.05) was observed in the group that had received the highest amount of PSO (38.97). The addition of PSO to the diet had a quadratic effect on redness index (a*) (8.26-9.09) of breast meat (p=0.042), but not of thigh meat (p&gt;0.05). Moreover, increasing amounts of PSO in the diet caused a gradual rise (p&lt;0.05) in the yellowness index (b*) in the thigh.</p>

			<p>According to Table 5, breast meat’s instrumental parameters had the following minimum and maximum values: pH (5.61-5.74), cooking losses (9.06-11.06 %), and drip losses (13.83-14.05 %). For the instrumental parameters of the thigh meat, the minimum and maximum values were as follows: pH (6.17-6.29), cooking losses (6.62-14.96%), and drip losses (14.17-15.76 %). As shown in Table 5, the pH of breast and thigh meat was higher for the control diet compared to the PSO diet. However, those differences were not significant (p&gt;0.05). In the group supplemented with 100 mg kg<sup>-1</sup> of PSO, breast cooking losses were reduced (p=0.022) compared to the control group (9.06 <em>vs.</em> 11.06). The highest cooking losses in thigh meat were found for the 200 mg kg<sup>-1</sup> PSO treatment, compared to the control and 100 mg kg<sup>-1</sup> PSO treatments. Regarding drip losses, no differences were observed between the study groups for either thigh or breast meat (p&gt;0.05).</p>

		</sec><!--/sec 3-->



		<table-wrap id="t5">
				<label>Table 5</label>
					<caption>
						<title>Instrumental quality parameters of breast and thigh meat from dietary experimental groups.</title>
					</caption>
					<table>
						<thead>
							<tr>
								<th align="center" rowspan="2">Parameters</th>
								<th align="center" colspan="3">PSO (mg kg<sup>-1</sup>)</th>
								<th align="center" rowspan="2">SEM</th>
								<th align="center" colspan="3">p-values</th>
							</tr>
							<tr>
								
								<th align="center">0</th>
								<th align="center">100</th>
								<th align="center">200</th>
								
								<th align="center">Anova</th>
								<th align="center">Linear</th>
								<th align="center">Quadratic</th>
							</tr>							
						</thead>
						<tbody>
							<tr>
								<td><strong>Breast</strong></td>
								<td align="center"></td>
								<td align="center"></td>
								<td align="center"></td>
								<td align="center"></td>
								<td align="center"></td>
								<td align="center"></td>
								<td align="center"></td>
							</tr>
							<tr>
								<td>pH</td>
								<td align="center">5.74</td>
								<td align="center">5.61</td>
								<td align="center">5.67</td>
								<td align="center">0.047</td>
								<td align="center">0.185</td>
								<td align="center">0.136</td>
								<td align="center">0.120</td>
							</tr>
							<tr>
								<td>Cooking losses (%)</td>
								<td align="center">11.06<sup>a</sup></td>
								<td align="center">9.06<sup>b</sup></td>
								<td align="center">9.71<sup>ab</sup></td>
								<td align="center">0.444</td>
								<td align="center">0.022</td>
								<td align="center">0.052</td>
								<td align="center">0.031</td>
							</tr>
							<tr>
								<td>Drip losses (%)</td>
								<td align="center">13.83</td>
								<td align="center">13.88</td>
								<td align="center">14.05</td>
								<td align="center">0.719</td>
								<td align="center">0.974</td>
								<td align="center">0.830</td>
								<td align="center">0.946</td>
							</tr>
							<tr>
								<td><strong>Thigh</strong></td>
								<td align="center"></td>
								<td align="center"></td>
								<td align="center"></td>
								<td align="center"></td>
								<td align="center"></td>
								<td align="center"></td>
								<td align="center"></td>
							</tr>
							<tr>
								<td>pH</td>
								<td align="center">6.29</td>
								<td align="center">6.17</td>
								<td align="center">6.20</td>
								<td align="center">0.058</td>
								<td align="center">0.391</td>
								<td align="center">0.312</td>
								<td align="center">0.357</td>
							</tr>
							<tr>
								<td>Cooking losses (%)</td>
								<td align="center">7.13<sup>b</sup></td>
								<td align="center">6.62<sup>b</sup></td>
								<td align="center">14.96<sup>a</sup></td>
								<td align="center">0.589</td>
								<td align="center">0.001</td>
								<td align="center">0.001</td>
								<td align="center">0.001</td>
							</tr>
							<tr>
								<td>Drip losses (%)</td>
								<td align="center">15.76</td>
								<td align="center">14.17</td>
								<td align="center">14.43</td>
								<td align="center">0.48</td>
								<td align="center">0.080</td>
								<td align="center">0.074</td>
								<td align="center">0.142</td>
							</tr>
						</tbody>
					</table>
					<table-wrap-foot>
						<fn id="TFN4">
							<p><sup>a,b</sup> Different superscripts indicate significant differences within a row (p &lt;0.05). PSO: pomegranate seed oil. SEM: standard error means.</p>
						</fn>
					</table-wrap-foot>
				</table-wrap>




			<table-wrap id="t4">
				<label>Table 4</label>
					<caption>
						<title>Colour (L*, a*, b*) of breast and thigh meat from dietary experimental groups.</title>
					</caption>
					<table>
						<thead>
							<tr>
								<th align="center" rowspan="2">Parameters</th>
								<th align="center" colspan="3">PSO (mg kg<sup>-1</sup>)</th>
								<th align="center" rowspan="2">SEM</th>
								<th align="center" colspan="3">p-values</th>
							</tr>
							<tr>
								
								<th align="center">0</th>
								<th align="center">100</th>
								<th align="center">200</th>
								
								<th align="center">Anova</th>
								<th align="center">Linear</th>
								<th align="center">Quadratic</th>
							</tr>							
						</thead>
						<tbody>
							<tr>
								<td><strong>Breast</strong></td>
								<td align="center"></td>
								<td align="center"></td>
								<td align="center"></td>
								<td align="center"></td>
								<td align="center"></td>
								<td align="center"></td>
								<td align="center"></td>
							</tr>
							<tr>
								<td>Lightness (L*)</td>
								<td align="center">36.20<sup>b</sup></td>
								<td align="center">39.14<sup>a</sup></td>
								<td align="center">39.07<sup>a</sup></td>
								<td align="center">0.563</td>
								<td align="center">0.004</td>
								<td align="center">0.004</td>
								<td align="center">0.050</td>
							</tr>
							<tr>
								<td>Redness index (a)*</td>
								<td align="center">8.28</td>
								<td align="center">9.09</td>
								<td align="center">8.26</td>
								<td align="center">0.2932</td>
								<td align="center">0.116</td>
								<td align="center">0.967</td>
								<td align="center">0.042</td>
							</tr>
							<tr>
								<td>Yellowness index (b*)</td>
								<td align="center">13.50<sup>b</sup></td>
								<td align="center">15.42<sup>a</sup></td>
								<td align="center">14.69<sup>a</sup></td>
								<td align="center">0.214</td>
								<td align="center">0.001</td>
								<td align="center">0.002</td>
								<td align="center">0.001</td>
							</tr>
							<tr>
								<td><strong>Thigh</strong></td>
								<td align="center"></td>
								<td align="center"></td>
								<td align="center"></td>
								<td align="center"></td>
								<td align="center"></td>
								<td align="center"></td>
								<td align="center"></td>
							</tr>
							<tr>
								<td>Lightness (L*)</td>
								<td align="center">36.87<sup>ab</sup></td>
								<td align="center">36.24<sup>b</sup></td>
								<td align="center">38.97<sup>a</sup></td>
								<td align="center">0.606</td>
								<td align="center">0.019</td>
								<td align="center">0.030</td>
								<td align="center">0.043</td>
							</tr>
							<tr>
								<td>Redness index (a)*</td>
								<td align="center">6.78</td>
								<td align="center">7.18</td>
								<td align="center">6.65</td>
								<td align="center">0.195</td>
								<td align="center">0.183</td>
								<td align="center">0.652</td>
								<td align="center">0.078</td>
							</tr>
							<tr>
								<td>Yellowness index (b*)</td>
								<td align="center">11.79<sup>b</sup></td>
								<td align="center">12.51<sup>ab</sup></td>
								<td align="center">12.82<sup>a</sup></td>
								<td align="center">0.243</td>
								<td align="center">0.030</td>
								<td align="center">0.011</td>
								<td align="center">0.500</td>
							</tr>
							
						</tbody>
					</table>
					<table-wrap-foot>
						<fn id="TFN4">
							<p><sup>a,b</sup> Different superscripts indicate significant differences within a row (p &lt;0.05). PSO: pomegranate seed oil. SEM: standard error means.</p>
						</fn>
					</table-wrap-foot>
				</table-wrap>




		<table-wrap id="t3">
				<label>Table 3</label>
					<caption>
						<title>Effect of pomegranate seed oil (PSO) supplementation to male quails’ (70-140 days of age) diet on carcass traits.</title>
					</caption>
					<table>
						<thead>
							<tr>
								<th align="center" rowspan="2">Parameters</th>
								<th align="center" colspan="3">PSO (mg kg<sup>-1</sup>)</th>
								<th align="center" rowspan="2">SEM</th>
								<th align="center" colspan="3">p-values</th>
							</tr>
							<tr>
								
								<th align="center">0</th>
								<th align="center">100</th>
								<th align="center">200</th>
								
								<th align="center">Anova</th>
								<th align="center">Linear</th>
								<th align="center">Quadratic</th>
							</tr>							
						</thead>
						<tbody>
							<tr>
								<td>Carcass yield <sup>[1,2]</sup></td>
								<td align="center">59.98</td>
								<td align="center">59.39</td>
								<td align="center">60.38</td>
								<td align="center">0.823</td>
								<td align="center">0.700</td>
								<td align="center">0.739</td>
								<td align="center">0.447</td>
							</tr>
							<tr>
								<td>Abdominal fat <sup>[1]</sup></td>
								<td align="center">1.66</td>
								<td align="center">1.72</td>
								<td align="center">1.21</td>
								<td align="center">0.312</td>
								<td align="center">0.474</td>
								<td align="center">0.333</td>
								<td align="center">0.465</td>
							</tr>
							<tr>
								<td>Breast <sup>[3]</sup></td>
								<td align="center">54.24</td>
								<td align="center">53.04</td>
								<td align="center">54.28</td>
								<td align="center">0.867</td>
								<td align="center">0.533</td>
								<td align="center">0.975</td>
								<td align="center">0.272</td>
							</tr>
							<tr>
								<td>Thigh and drumstick <sup>[3]</sup></td>
								<td align="center">36.63</td>
								<td align="center">36.53</td>
								<td align="center">34.86</td>
								<td align="center">0.690</td>
								<td align="center">0.169</td>
								<td align="center">0.096</td>
								<td align="center">0.371</td>
							</tr>
							<tr>
								<td>Liver <sup>[1]</sup></td>
								<td align="center">1.33</td>
								<td align="center">1.43</td>
								<td align="center">1.36</td>
								<td align="center">0.101</td>
								<td align="center">0.779</td>
								<td align="center">0.848</td>
								<td align="center">0.506</td>
							</tr>
							<tr>
								<td>Heart <sup>[1]</sup></td>
								<td align="center">0.79</td>
								<td align="center">0.82</td>
								<td align="center">0.81</td>
								<td align="center">0.036</td>
								<td align="center">0.830</td>
								<td align="center">0.633</td>
								<td align="center">0.716</td>
							</tr>
							<tr>
								<td>Pancreas <sup>[1]</sup></td>
								<td align="center">0.17</td>
								<td align="center">0.17</td>
								<td align="center">0.18</td>
								<td align="center">0.006</td>
								<td align="center">0.178</td>
								<td align="center">0.269</td>
								<td align="center">0.129</td>
							</tr>
						</tbody>
					</table>
					<table-wrap-foot>
						<fn id="TFN3">
							<p><sup>[1]</sup> Calculated as a percentage of live body weight. <sup>[2]</sup> Carcass yield was obtained by separating the head, feet, feathers, digestive organs and giblets. <sup>[3]</sup> Calculated as a percentage of carcass weight. SEM: standard error means.</p>
						</fn>
					</table-wrap-foot>
				</table-wrap>




		<table-wrap id="t2">
				<label>Table 2</label>
					<caption>
						<title>Effect of pomegranate seed oil (PSO) supplementation to male quails’ (70-140 days of age) diet on performance parameters.</title>
					</caption>
					<table>
						<thead>
							<tr>
								<th align="center" rowspan="2">Parameters</th>
								<th align="center" colspan="3">PSO (mg kg<sup>-1</sup>)</th>
								<th align="center" rowspan="2">SEM</th>
								<th align="center" colspan="3">p-values</th>
							</tr>
							<tr>
								
								<th align="center">0</th>
								<th align="center">100</th>
								<th align="center">200</th>
								
								<th align="center">Anova</th>
								<th align="center">Linear</th>
								<th align="center">Quadratic</th>
							</tr>							
						</thead>
						<tbody>
							<tr>
								<td>Initial body weight (g)</td>
								<td align="center">183.00</td>
								<td align="center">180.70</td>
								<td align="center">178.60</td>
								<td align="center">6.015</td>
								<td align="center">0.876</td>
								<td align="center">0.614</td>
								<td align="center">0.989</td>
							</tr>
							<tr>
								<td>Final body weight (g)</td>
								<td align="center">213.70</td>
								<td align="center">209.80</td>
								<td align="center">194.60</td>
								<td align="center">5.903</td>
								<td align="center">0.093</td>
								<td align="center">0.041</td>
								<td align="center">0.450</td>
							</tr>
							<tr>
								<td>Body weight gain (g)</td>
								<td align="center">30.70<sup>a</sup></td>
								<td align="center">29.10<sup>ab</sup></td>
								<td align="center">16.00<sup>b</sup></td>
								<td align="center">3.492</td>
								<td align="center">0.022</td>
								<td align="center">0.012</td>
								<td align="center">0.204</td>
							</tr>
							<tr>
								<td>Feed intake (g/period/quail)</td>
								<td align="center">693.10<sup>a</sup></td>
								<td align="center">680.80<sup>ab</sup></td>
								<td align="center">610.10<sup>b</sup></td>
								<td align="center">20.813</td>
								<td align="center">0.032</td>
								<td align="center">0.015</td>
								<td align="center">0.274</td>
							</tr>
						</tbody>
					</table>
					<table-wrap-foot>
						<fn id="TFN2">
							<p><sup>a,b;</sup> Different superscripts indicate significant differences within a row (p &lt; 0.05). SEM: standard error </p>
						</fn>
					</table-wrap-foot>
				</table-wrap>

		<sec id="sec4" sec-type="results">
			<title>Discussion</title>

			<p>Pomegranate by-products may have the potential to be a good source of nutrients and antioxidants for livestock nutrition (Emami et al., 2015). In the current experiment, PSO had an impact on some performance parameters such as body weight gain and FI. In addition, it was observed that levels of 200 mg kg<sup>-1</sup> of PSO resulted in the lowest value of final body weight and abdominal fat. Although these differences were not significant (Table 3), a trend was observed for final body weight. Similar to the results of the current study, adding PSO to high-fat diets decreased body weight, body weight gain, fat, and biomarkers of cholesterol profile in rats (McFarlin et al., 2008; Vroegrijk et al., 2011). These authors proposed PSO supplementation in the diet would reduce leptin and increase adiponectin concentration, leading to lower weight and body fat content. Furthermore, McFarlin et al. (2008) speculated that PSO supplementation can alter the way that dietary fat interacts with the brain. These authors suggested that PSO could increase fat metabolism, resulting in the attenuation of weight gain that they observed. However, further research is needed to explain the effect of PSO on pathways that mediate body weight gain. These results were in disagreement with Szymczyk &amp; Szczurek (2016) who have shown that body weight gain was not affected by PSO (from 5000 to 15000 mg kg<sup>-1</sup>) supplementation to the diet in broilers, while Saleh et al. (2016) mentioned the addition of 100, 200, and 300 mg kg<sup>-1</sup> of pomegranate peel extract or 1, 2, and 3 g kg<sup>-1</sup> of pomegranate peel to the diet resulted in the reduction of growth performance for broilers up to 42 days. Nekooeian et al. (2014) showed that 200 or 600 mg kg<sup>-1</sup> PSO per day did not cause significant changes in the body weight of diabetic rats. Probably, those differences could be attributed to the different animal species included in the trials and the total amount of PSO included in the diet.</p>

			<p>In addition, this study showed that FI decreased linearly as dietary PSO levels increased. This fact seems likely to be related to the palatability of the feed. The decrease in FI also affected the final body weight and body weight change of male quails. Pomegranate is a rich source of hydrolysable tannins, although the content of tannins is lower in the seed (Bar-Ya’akov et al., 2019). The reduction in FI can be explained by reduced diet palatability due to the presence of tannin in the PSO (Gungor et al., 2021) and body weight gain and final body weight could be affected. In agreement with these results, the lowest FI was observed by previous authors at the highest concentration of PSO in laying hens (Kostogrys et al., 2017) and broilers (Gungor et al., 2021). Rezvani et al. (2018) found that PSO negatively affected FI and growth performance, especially in young broilers. There is a possibility that these results are due to the amount of pomegranate seed extract provided in the diet and that the animals may not be able to adapt to the new diets.</p>

			<p>Dressed poultry carcasses, which are dead poultry bodies after being partially butchered without internal organs, heads, and legs, are influenced by both genetic and environmental factors including age, sex, line, and livestock management (Ali et al., 2019). The values obtained for carcass yield agree with those published by Ali et al. (2019) which are between 55.73 and 58.86 (%). Conversely, these results were relatively lower than those published by Raji et al. (2015) for mature Japanese quail, which can reach 89.82%. Several factors may contribute to this difference, including genetics and environment. In addition, carcass percentage may be directly affected by live body weight. A higher body weight of quail results in a higher dressed carcass weight and carcass percentage. These findings are consistent with those observed in our study, which showed a lower final weight of quails. The values obtained for breast percent (54.28 to 53.04%) were higher than those found by Raji et al. (2015), although these authors evaluated deboned breasts in their study. In line with the current findings are those published by previous authors for bone-in breast, which range from 40 to 50% (Alkan et al., 2012; Lukanov et al., 2018; Hussen, 2020). Similar results are observed for the thigh value, which was higher in the current study because the thigh and drumstick were evaluated together. The rest of the parameters evaluated (including organs and fat) are in line with those shown by Raji et al. (2015) for Japanese quails. However, slight differences can be found which, according to these authors, could be justified by the genetics of the quail. Quail meat with excellent dressed carcass characteristics could be produced by crossbreeding quails that are genetically superior to quail meat.</p>

			<p>Despite the reduction in final body weight, no changes were observed in the slaughter yield of quails with the dietary addition of PSO at 100 and 200 mg kg<sup>-1</sup>. Moreover, in the current study proportions of breast, thigh, drumstick, liver, heart, and pancreas in the carcasses did not differ among the groups. Also, the findings of the current study are consistent with those get by Szymczyk &amp; Szczurek (2016) who replaced soybean oil with PSO in chicken diets. Those authors did not find significant differences in carcass yield, breast and leg muscles, liver, gizzard, and heart weights of chickens. The only exception was an abdominal fat percentage, which was found to be higher in broilers fed a diet containing 1500 mg kg<sup>-1</sup> PSO. Banaszkiewicz et al. (2018) did not find any influence of PSO supplementation in the slaughter yield, proportions of breast, thigh, total muscle, and abdominal fat. In the same trend, Jankowski et al. (2012) observed no differences in slaughter yield or breast, thigh, and drumstick muscle yield in turkeys fed diets containing soybean, linseed, and rapeseed oils. In partial agreement with our feedback Hamady et al. (2015) proved that the supplementation of pomegranate peel powder did not influence liver, gizzard, or heart weight percentages of broiler chickens. However, those authors noticed that the addition of pomegranate peel powder to the diet increased thigh and breast weight percentages. The findings of the current study on carcass traits disagree with Akuru et al. (2021), who reported the hat thigh, breast, and heart weight of broilers improved with pomegranate peel powder supplementation in the diet. Those differences are probably due to the different compositions of the pomegranate co-product included in the diet.</p>

			<p>Colour is one of the important quality traits influencing consumer preferences (Vieira et al., 2021). The addition of PSO led to significant differences in the colour parameters. Increased lightness (L*) was found in breast and thigh meat as PSO levels increased. According to Abdel Baset et al. (2022), meat weight might decrease due to reduced water-holding capacity (WHC), resulting in lighter meat, which is consistent with what was observed in this study. Moreover, an increase in yellowness index (b*) was observed in the breast and thigh when PSO was added to the diet. In different studies, pomegranate coproducts had a colour stabilizing effect on muscle food. However, the results are not conclusive. For example, Devatkal et al. (2010) found that the incorporation of pomegranate rind powder was effective in reducing L-values, but no differences were observed among pomegranate seed powder and control for L values. Conversely, redness was reduced when using pomegranate rind powder and pomegranate seed powder compared with the control goat patties. Similar results have been described in frankfurter samples containing white pomegranate juice concentrate, red pomegranate juice concentrate, and pomegranate rind powder extract (Das et al., 2021). It was observed that the inclusion of these ingredients reduced the L* and a* values concerning to the control. In a similar way, Serdaroğlu et al. (2021) reported a decrease in these values when pomegranate seed powder was used in chicken meat emulsion. Contrary, Gungor et al. (2021) did not find any difference in breast colour (in terms of lightness, yellowness or redness) among different PSO diets. Pomegranate is a good source of bioactive substances such as tocopherol and xanthophyll (Madrigal-Carballo et al., 2009). The consumption of natural ingredients rich in these substances has been linked to changes in meat colour. Processing and storing of these ingredients may alter their oxidation levels, resulting in a change in FI and consequently, a change in xanthophyll deposition in meat (Pertiwi et al., 2022). The differences in meat colour found among the studies consulted could be justified for this reason.</p>

			<p>According to Terlouw et al. (2008) normal pH of quail breast meat ranged between 5.4 and 6.0. The pH in the current study was on average 5.67, regardless of the diet received, which can be characterized as normal breast pH. On the other hand, the values for thigh pH (6.22) were slightly lower than those reported by Nasr et al. (2017), which ranged between 6.24-6.37. Those differences could be attributed to slaughter age, strain, or line. The use of PSO in the current study did not significantly affect the pH of breast and thigh muscles, but the addition of PSO numerically decreased the pH in the breast and thigh muscles. These results are in line with those published by Banaszkiewicz et al. (2018), who showed that the use of PSO in a broiler diet did not influence meat pH. Likewise, Akuru et al. (2021) pointed out that dietary supplementation of pomegranate peel powder meal at four levels (2, 4, 6 and 8 g/kg) did not influence the pH of the broiler breast meat compared to the control diet.</p>

			<p>Water holding capacity is determined as drip losses and cooking losses. The lowest value of cooking losses in the breast was found for the 100 mg kg<sup>-1</sup> dose. However, contrary to expectations, the value of cooking losses was increased in the thigh of quails that had received 200 mg kg<sup>-1</sup> of PSO. Kaur et al. (2015) reported that with the increase in the ratio of pomegranate seed powder added to chicken nuggets, there was a significant decrease in emulsion stability and cooking efficiency. The authors propose a decrease in WHC as a consequence of lower pH. In contrast, Keşkekoğlu &amp; Üren (2014) described that there were no significant differences between the controls and the pomegranate seed extract containing meatball samples, which is consistent with those described by Kurt (2017) for beef patties. Moreover, a trend was observed for drip losses, which decreased as the concentration of PSO in the diet increased. Similar results had been reported by Emami et al. (2015), who showed that the inclusion of pomegranate seed powder in a goat diet decreased drip loss of meat. Those authors suggest a negative correlation between intramuscular fat content and drip losses. Intramuscular fat may act as ‘insulation’ and reduce water loss in meat. The same behaviour was observed by several authors, who proposed that the incorporation of pomegranate coproducts in meat enhances the WHC of raw beef sausage (Gullón et al., 2020), beef meatballs (Morsy et al., 2018), and beef burgers (Abdel-Fattah et al., 2016). However, since our results are not conclusive for WHC in the different parts of the carcass evaluated, further research is required.</p>

			<p>In summary, pomegranate seed oil supplementation at a level of 200 mg kg<sup>-1</sup> reduced FI and body weight gain without affecting carcass traits. Compared to the control, PSO supplementation increased L* and b* in the thigh and breast meat. Likewise, dietary PSO at 100 mg kg<sup>-1</sup> significantly improved some characteristics of cooked meat. According to these findings, the inclusion of PSO at 100 mg kg<sup>-1</sup> level in the quails can be a promising method to increase some characteristics of meat quality, but a higher concentration has detrimental effects on quail performance and meat quality. Further research is needed to find the optimal dose of PSO for quails to improve these parameters.</p>

		</sec><!--/sec 4-->				

					
		
	</body>
	<back>
		<author-notes>
			<title>Authors’ contributions</title>
			<fn>Conceptualization: A Sarmiento-García, B Sevim, O Olgun.</fn>
			<fn>Data curation: A Sarmiento-García, B Sevim, O Olgun.</fn>
			<fn>Formal analysis: A Sarmiento-García, B Sevim, O Olgun.</fn>
			<fn>Funding acquisition: B Sevim, O Olgun.</fn>
			<fn>Investigation: B Sevim, O Olgun, SA Gökmen.</fn>
			<fn>Methodology: A Sarmiento-García, B Sevim, O Olgun.</fn>
			<fn>Project administration: B Sevim, O Olgun, SA Gökmen.</fn>
			<fn>Resources: A Sarmiento-García, B Sevim, O Olgun.</fn>
			<fn>Software: SA Gökmen.</fn>
			<fn>Supervision: A Sarmiento-García, B Sevim, O Olgun.</fn>
			<fn>Validation: A Sarmiento-García, B Sevim, O Olgun.</fn>
			<fn>Visualization: A Sarmiento-García, B Sevim, O Olgun.</fn>
			<fn>Writing – original draft: A Sarmiento-García.</fn>
			<fn>Writing – review &amp; editing: A Sarmiento-García, B Sevim, O Olgun, SA Gökmen.</fn>
		</author-notes>

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