Journal of Animal Science and Technology
Korean Society of Animal Sciences and Technology
RESEARCH ARTICLE

Influence of ferulic acid and clinoptilolite supplementation on growth performance, carcass, meat quality, and fatty acid profile of finished lambs

Ana Tánori-Lozano1https://orcid.org/0000-0002-3180-5122, Adrián Imanol Quintana-Romandía1https://orcid.org/0000-0002-2071-8488, Maricela Montalvo-Corral1https://orcid.org/0000-0002-0070-7490, Araceli Pinelli-Saavedra1https://orcid.org/0000-0003-1487-5767, Martín Valenzuela-Melendres1https://orcid.org/0000-0002-7347-9599, José Luis Dávila-Ramírez2https://orcid.org/0000-0002-4297-5426, Thalia Yamileth Islava-Lagarda1https://orcid.org/0000-0001-8404-2969, Humberto González-Ríos1,*https://orcid.org/0000-0002-7463-778X
1Centro de Investigación en Alimentación y Desarrollo, A.C. (CIAD, A.C.), Hermosillo, Sonora 83304, Mexico
2Ciencia Aplicada para el Desarrollo Tecnológico, A.C. (CIADETEC, A.C.), Hermosillo, Sonora 83260, Mexico
*Corresponding author: Humberto González-Ríos, Centro de Investigación en Alimentación y Desarrollo, A.C. (CIAD, A.C.), Hermosillo, Sonora 83304, Mexico. Tel: +52-662-289-2400, E-mail: hugory@ciad.mx

© Copyright 2022 Korean Society of Animal Science and Technology. This is an Open-Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Received: Nov 13, 2021; Revised: Jan 19, 2022; Accepted: Mar 15, 2022

Published Online: Mar 31, 2022

Abstract

This study was conducted to evaluate the effect of ferulic acid (FA) and clinoptilolite (CTL) supplementation on the growth performance, carcass characteristics, and meat quality of hair-breed lambs. Twenty-eight Kathadin male lambs (33.72 ± 3.4 kg) were randomly allocated to one of the four diets (n=7) under a 2 × 2 factorial arrangement to evaluate the effect of FA (0 or 300 ppm) and CTL (0% or 1%) during the last 40 days of the finishing phase. No interaction between additives was shown for growth performance, carcass characteristics and meat quality, with exception of the fatty acid profile (p < 0.05). FA reduced feed intake and carcass conformation (p < 0.05). Wholesale cuts were not affected by FA or CTL (p > 0.05). The L*, a*, and C* color parameters and some intramuscular fatty acids of the longissimus thoracis muscle were positively modified by CTL supplementation (p < 0.05). While there was no FA × CTL interaction, each additive could be used individually in animal nutrition to improve the feedlot performance and meat quality of the lambs.

Keywords: Phytochemicals; Zeolite; Sheep; Carcass evaluation; Feedlot performance

INTRODUCTION

As a strategy to improve the meat production chain, it is common to use synthetic growth promoters as anabolic agents to enhance animal performance and carcass characteristics [1]. However, synthetic compounds are becoming unfeasible due to consumer concerns and strict regulations in Europe [2], and natural alternatives are more acceptable for use in animal production. The phytochemicals dietary supplementation could be a strategy to improve animal performance with the same beneficial effect as synthetic growth promoters but without compromising animal welfare, meat quality, and consumer health [2-4]. Recently, some researchers have reported that ferulic acid (FA; a secondary plant compound with bioactive properties) enhances growth performance, carcass characteristics, and meat quality in ruminants by reducing lipid peroxidation and increasing the average daily gain (ADG) and carcass weight [5-7]. Nevertheless, growth performance and carcass characteristics after FA supplementation in lambs have been inconsistent; in addition, the optimal dose or time exposition has not been established [8-11]. On the other hand, some authors suggest that the moderate growth-promoting effect of phytochemicals such as FA, in the case of ruminants, could be related to their antimicrobial activity, which causes a productive improvement due to the modification of ruminal fermentation [4,12]. However, more studies are required to establish the possible mechanisms of action involved.

On the other hand, clinoptilolite (CTL) a naturally occurring zeolite, is classified as safe and is commercially available for use in diets for broilers and pigs, but it is uncommon to use it in beef cattle production [13,14]. A study has shown that supplementing lambs with CTL increased muscle tissue deposition and decreased fat tissue deposition [15]. Additionally, an improvement in feed efficiency and ADG was reported [16,17]; the ion-exchange property of CTL favorably modulates ruminal fermentation, improving energy efficiency, which is reflected in animal growth performance [18]. However, the available results are conflicting; hence, more research is required in order to fully elucidate CTL’s response in ruminants.

There is a lack of studies reporting the effects of FA or CTL on the growth performance and meat quality of lambs. Moreover, the combined effect of both additives has not been studied. Therefore, this research hypothesis was that the simultaneous administration of dietary FA and CTL to hair lambs could synergistically enhance the growth performance, carcass characteristics, and meat quality. This study aims to evaluate the combined effect of the dietary inclusion of FA and CTL on the growth performance, carcass characteristics, meat quality, and chemical composition of finished hair-breed lambs.

MATERIALS AND METHODS

Study site

The feedlot performance was carried out during the winter season at a commercial sheep production farm located in northwestern Sonora, Mexico (latitude 28.78°N and longitude 111.40°W). The average temperature and relative humidity during the study were 18 ± 12°C and 49 ± 14%, respectively. The animal slaughter and carcass evaluation were conducted at the Agriculture and Livestock Department (ALD) of the Universidad de Sonora (UNISON), situated 21 km from Hermosillo, Sonora, Mexico. Meat quality, chemical composition, and fatty acid profile were evaluated at the Centro de Investigación en Alimentación y Desarrollo (CIAD), also situated in Hermosillo, Sonora.

All animal management and slaughter procedures were conducted in accordance with the official techniques and standards in Mexico (NOM-051-ZOO-1995, NOM-033-ZOO-1995, and NOM-062-ZOO-1999). Furthermore, the Ethics Committee of CIAD accepted and supervised all of the experimental processes (CEI/002-2/2021).

Animals and treatments

Twenty-eight Katahdin male lambs with similar initial body weight (IBW; 33.74 ± 3.4 kg) were individually housed in pens (2.5 m × 2 m) for this experiment. Seven pens per group were randomly assigned to one of the four experimental diets as follows: (1) control (high concentrate basal diet; BD); (2) FA (BD with 300 ppm FA; Laboratorios Minkab, Guadalajara, Jalisco, Mexico); (3) CTL (BD with 1% CTL; Zeolex®; Grupo Sanfer, Ciudad de Mexico, Mexico); and (4) FAZ (BD with 300 ppm FA + 1% CTL). All animals were given a compound of vitamins (A, D, and E) at the beginning of the study and were treated with Ivermectin (ivermectin + ADE; Virbamec ADE FUERTE; Virbac Mexico, Zapopan, Jalisco, Mexico; 0.7 mL/animal) for eliminating internal and external parasites. Also, each lamb was weighed and identified with a numbered plastic ear tag before being assigned to the experimental diets.

The feeding period was 50 days, ten days for the BD adaptation (12.9% crude protein [CP], 3.2 Mcal/kg feed of metabolizable energy [ME], meeting the recommended nutritional requirements for meat-producing lambs) [19] and the next forty days for the experimental trial. To ensure the daily intake of FA and CTL per lamb, both were mixed with mineral premix and subsequently blended with other ground ingredients to create the corresponding FA or CTL dose (300 ppm or 1%). Finally, the concentrate mixture was incorporated with the remaining ingredients. The ingredients and chemical composition of the experimental diets are indicated in Table 1. Feed was offered ad libitum twice a day at 0800 h and 1600 h in half mixed rations, and the lambs had ad libitum access to fresh water. Additionally, their health status was monitored daily by direct visual examination.

Table 1. Ingredients and chemical composition of experimental diets offered to the hair lambs
Item Treatment1)
Control FA CTL FAZ
Ingredients (% of DM)
 Wheat straw 10.2 10.2 10 10
 Alfalfa hay 12 12 12.1 12.1
 Corn grain 61 61 60.1 60.1
 Poultry manure 5 5 5 5
 Soybean meal 8 8 8 8
 Cane molasses 3 3 3 3
 Mineral premix 0.5 0.5 0.5 0.5
 Salt 0.3 0.3 0.3 0.3
 Ferulic acid (ppm) 0 300 0 300
 Zeolite (Clinoptilolite) 0 0 1 1
Chemical composition (DM basis)
 Dry matter (%) 88.5 88.5 88.5 88.5
 CP (%) 12.9 12.9 12.8 12.8
 Ether extract (%) 3.60 3.60 3.61 3.61
 Metabolizable energy2) (Mcal/kg) 3.25 3.25 3.21 3.21
 NEm2) (Mcal/ kg) 2.18 2.18 2.15 2.15
 NEg2) (Mcal/ kg) 1.44 1.44 1.42 1.42

Control, high concentrate basal diet (BD); FA, BD with 300 ppm ferulic acid; CTL, BD with 1% clinoptilolite; FAZ, BD with 300 ppm FA + 1% CTL.

Based on tabular energy values of ingredients from NRC [19].

DM, dry matter; CP, crude protein; NEm, net energy for maintenance; NEg, net energy for gain.

Download Excel Table
Feedlot performance

Animals were individually weighed three times (initial, interim, and final) during the feeding period. From these data, ADG was calculated for the first period (d 0 to 20), the second period (d 21 to 40), and for the whole period (d 0 to 40). Feed intake (feed offered – refusal) was measured daily. At the beginning of the experimental period, dry matter intake was established at 5% of the live weight, and then, the daily feed ration was adjusted to have a minimum refusal (< 10%). Finally, feed efficiency was indicated as the ratio of daily feed intake to ADG.

Carcass characteristics, longissimus thoracis muscle sampling, and wholesale cut yields

At the end of the experimental period, the lambs were slaughtered (ALD slaughterhouse) after fasting for 16 h (with access to water). Then, once the lambs were skinned and eviscerated, their carcasses were weighed to determine hot carcass weight (HCW); after chilling the carcasses for 24 h at 4°C, the cold carcass weight (CCW) was measured. Subsequently, following the methodology described byAMSA [20], carcass conformation was evaluated (numeric scale: 1 = bad to 8 = excellent). The cooling loss was also estimated by the difference between HCW and CCW expressed as a percentage. Moreover, body measurements were registered: carcass length, thorax depth, leg length, and leg perimeter [21].

Then, the carcasses were divided along the midline, and each left side was ribbed between the 12th and 13th ribs to measure the back-fat thickness and loin area using a dot squared grid. Finally, half of the carcasses were weighed and split into forequarters and hindquarters to obtain wholesale cuts: neck, shoulder, loin, ribs and flank, plain loin, and leg [22]. Each cut was expressed as a percentage of the half carcass weight.

Meat quality evaluation

At 24 h postmortem, the pH from the longissimus thoracis (LT) was directly measured between the 12th and 13th ribs with a portable pH meter (model HI 99163, Hanna Instruments, Wilmington, MA, USA) with a puncture electrode. Then, after splitting and weighing each wholesale cut, the LT muscle was dissected from the loin of each left side carcass (located at the top of the 4th and 12th ribs). Subsequently, each muscle sample was vacuum packed, stored, and aged for one week at −18°C in the Meat Science Laboratory of CIAD. At the end of the aging time, the samples were thawed (24 h at 0°C) and sectioned following the diagram shown in Fig. 1 to determine final pH, water-holding capacity (WHC), color values, cooking loss, fatty acid profile, and Warner–Bratzler shear force (WBSF).

jast-64-2-274-g1
Fig. 1. Longissimus thoracis muscle sampling diagram.
Download Original Figure

To evaluate the final meat pH, each steak was homogenized in distilled water with a meat: water ratio of 1:3 (g:mL). Then, the final meat pH was measured using a benchtop digital pH meter previously calibrated with pH 4 and 7 standards (Model HI-2210, Hanna Instruments Digital, Woonsocket, RI, USA). WHC was evaluated following the technique previously described by Sutton et al. [23]. Meat samples were centrifuged for a lapse of time (1,420 g × 5 min, at 5°). The WHC value was calculated by the difference between the initial weight (before centrifugation) and final weight (after centrifugation) of the sample, expressed as a percentage.

LT muscle color analysis was conducted after 30 min of blooming using a Minolta CR-2600 colorimeter with illuminant D65 (10° observer and an 8 mm diameter aperture; Konica Minolta Sensing, Osaka, Japan), where L*, a*, and b* were documented. Chroma (C*; color saturation index) and hue angle (H*) were calculated using the redness and yellowness values, according to the Cassens et al. [24] methodology. Five measures were taken directly from the LT muscle surface in distinct locations from each sample.

To evaluate the cooking loss and WBSF, LT samples were weighed raw, and after being cooked until the internal temperature of the steak reached 71°C (an electric skillet was employed; Cook Master 3222-3, Oster, Mississauga, Ontario, Canada), they were immediately weighed again. Next, the cooked samples were chilled, first, at room temperature (22°C–27°C), and then, at 4°C for 24 h. Finally, from each steak, six cores of 3 cm long and 1.27 cm2 in cross-section were cut parallel to the muscle fibers and used to evaluate WBSF with a WBSF device (TA. XT Plus Texture Analyzer texturometer, Texture Technologies, Scarsdale, NY, USA). The WBSF value was indicated in kg. The percentage of cooking loss was determined as the ratio of the raw weight to the cooked weight.

Chemical composition and fatty acid profile

Moisture, ash, fat, and protein contents of the LT muscle were determined as described by the AOAC standardized procedures [25]. To evaluate the fatty acid profile, lipid extraction was carried out following an adaptation to the Bligh and Dyer [26] procedure with the following modifications: 2 g of meat was mixed with 10 mL of methanol and homogenized (Ultra-Turrax T25, IKA, Staufen, Germany) for 1 minute at 1,450×g. Then, 20 mL of chloroform was added, homogenized again for 2 minutes at 1,450×g, and filtered with Whatman no. 41. Later, three milliliters of KCl (0.88%) was added, and after vigorous shaking, the top layer was poured out. Finally, 4 mL of distilled water and methanol were added to assist separation before the sample was shaken again; the top layer containing the non-lipid fractions was removed, and the lower phase was used for esterification.

Fatty acid methyl esters (FAMEs) were prepared following an adaptation of the standard method described by Li and Watkins [27]. First, samples were placed in a water bath (40°C) with a constant nitrogen gas flow until all solvents were evaporated. Then, 4 mL of NaOH (0.5 N in methanol) was added; later, tubes (previously capped) were placed in a water bath (90°C) for 5 min, and after cooling at room temperature, 5 mL of boron trifluoride (BF3) was added to the samples and heated in a water bath for 5 min again. Next, when the samples were cooled, 4 mL of hexane was added and then heated for 2 min in a water bath (90°C). Finally, 0.5 g of anhydrous sodium sulfate and 1 mL of saturated NaCl solution were added, and 1 mL of the top lipid phase was carefully collected into a 2 mL gas chromatography sample vial containing 1 mL of hexane.

FAMEs were analyzed using a gas chromatograph (Hewlett Packard model 6890, Waldbronn, Germany) equipped with a flame ionization detector and a capillary column (Agilent J&W DB-23 [0.25mm internal diameter x 60 m; 0.25 μm film thickness]). The chromatographic conditions were as previously described by González-Ríos et al. [5]. Identification of each FAME was based on the retention times compared with those from a known standard (tridecanoic acid; 13:0, Sigma-Aldrich, St Louis, MO, USA). Data are expressed as the percentage of total fatty acids identified. Nutritional ratios (polyunsaturated fatty acid [PUFA] / saturated fatty acid [SFA], monounsaturated fatty acid [MUFA] / SFA, and n-6 / n-3) were calculated as well as the total amounts of PUFA, MUFA, and SFA.

Statistical analysis

Data acquired from the study were analyzed with the NCSS statistical program (version 2007, NCSS, Kaysville, Utah, USA). Data from feedlot performance, carcass characteristics, wholesale cuts, meat quality, chemical composition, and fatty acid profile were analyzed as a 2 × 2 factorial under a complete randomized design. The model considered fixed effects of dietary supplementation of FA and CTL and their interaction (CTL × FA). For feedlot performance variables, IBW was included as a covariate in the model. Differences among means were compared using the Tukey–Kramer test. Significances were considered when P < 0.05, and a trend was declared when 0.05 < p < 0.10.

RESULTS

Feedlot performance

Table 2 presents the influence of dietary supplementation with FA and CTL on hair-breed lamb feedlot performance. No variable throughout the feeding trial or by period was affected by the FA × CTL interaction (p > 0.05). Neither IBW nor final body weight differed between experimental diets (p > 0.05). Additionally, ADG, feed intake, and feed conversion were not affected (p > 0.05) during the 40 d trial period. Lambs supplemented with CTL tended to increase their daily feed intake through each period by 7.5% (d 0 to 20; p = 0.088) and 7.6% (d 21 to 40; p = 0.054), while feed conversion tended (p = 0.065) to be higher in the second period; however, this trend was not shown during overall feeding period (d 0 to 40; p > 0.05). Moreover, from d 21 to 40, dietary FA decreased lamb daily feed intake by 7% (p < 0.05), but this reduction did not affect their feed conversion (p > 0.05).

Table 2. Feedlot performance of the hair lambs supplemented with FA and CTL
Item FA (ppm)1) CTL (%)2) SEM p-value
0 300 0 1 FA CTL FA × CTL
Initial body weight (kg) 34.10 33.39 33.78 33.70 0.97 0.619 0.956 0.619
Final body weight (kg) 45.55 45.39 45.57 45.37 0.65 0.866 0.833 0.673
Period 1 (d 0–20)
 Average daily gain (kg/d) 0.248 0.259 0.249 0.258 0.03 0.736 0.787 0.456
 Feed intake (kg/d, DM) 1.40 1.34 1.32 1.42 0.05 0.310 0.088 0.508
 Feed conversion (kg DM) 5.74 5.44 5.19 5.99 0.56 0.618 0.182 0.641
Period 2 (d 21–40)
 Average daily gain (kg) 0.343 0.330 0.349 0.324 0.03 0.708 0.444 0.740
 Feed intake (kg/d, DM) 1.69 1.57 1.57 1.69 0.05 0.039 0.054 0.995
 Feed conversion (kg DM) 4.54 4.91 4.35 5.11 0.37 0.357 0.065 0.488
Total (d 0–40)
 Average daily gain (kg/d) 0.295 0.291 0.296 0.291 0.02 0.866 0.833 0.673
 Feed intake (kg/d, DM) 1.52 1.46 1.45 1.53 0.05 0.264 0.142 0.809
 Feed conversion (kg DM) 5.30 5.25 5.06 5.49 0.46 0.913 0.381 0.545

Lambs fed 0 or 300 ppm of ferulic acid through the 40-d feeding trial.

Lambs fed 0 or 1% of clinoptilolite through the 40-d feeding.

FA, ferulic acid; CTL, clinoptilolite; DM, dry matter.

Download Excel Table
Carcass characteristics and wholesale cuts yield

Regarding the carcass characteristics, no significant FA × CTL interaction (p > 0.05) was shown (Table 3). Neither FA nor CTL had an effect on HCW, CCW, dressing weight, fat thickness, or loin area (p > 0.05; Table 3). However, the cooling loss value of the carcass from lambs supplemented only with CTL tended to be 47% lower (p = 0.072). In addition, dietary FA did affect the carcass conformation (p < 0.05); nevertheless, the conformation score was lower for this group (7.4%; Table 3). No FA × CTL interaction or FA main effect (p > 0.05) was presented for any of the body measurements (Table 3). On the other hand, while CTL tended (p = 0.057) to increase leg length (4%), other variables (carcass length, thorax depth, and leg perimeter) remained unaltered (p > 0.05). Wholesale cuts yield are presented in Table 4. Dietary FA, CTL, or their interaction (FA × CTL) did not impact (p > 0.05) any wholesale cut yield of the hair-breed lambs: neck, shoulder, loin, ribs and flank, plain loin, or leg.

Table 3. Carcass characteristics of the hair lambs supplemented with FA and CTL
Item FA (ppm)1) CTL (%)2) SEM p-value
0 300 0 1 FA CTL FA × CTL
HCW (kg) 22.46 21.91 22.33 22.03 0.49 0.304 0.573 0.164
CCW (kg) 22.00 21.50 21.83 21.68 0.49 0.356 0.776 0.134
Cooling loss (%) 1.15 1.11 1.48 0.78 0.35 0.910 0.072 0.325
Dressing weight (%) 48.13 47.13 47.92 47.34 0.95 0.328 0.573 0.565
Conformation (units) 6.46 5.98 6.32 6.12 0.20 0.036 0.355 0.583
Fat thickness (mm) 1.57 1.88 1.82 1.62 0.23 0.206 0.419 0.176
Loin area (cm2) 16.43 15.44 15.42 16.44 0.86 0.289 0.277 0.769
Body measurements (cm)
 Carcass length 64.13 64.92 65.06 64.00 1.38 0.592 0.475 0.859
 Thorax depth 18.38 18.30 18.44 18.24 0.31 0.805 0.557 0.879
 Leg length 38.68 37.57 37.35 38.91 0.72 0.161 0.057 0.305
 Leg perimeter 43.97 43.73 44.39 43.55 0.76 0.767 0.724 0.182

Lambs fed 0 or 300 ppm of ferulic acid through the 40-d feeding trial.

Lambs fed 0 or 1% of clinoptilolite through the 40-d feeding.

FA, ferulic acid; CTL, clinoptilolite; HCW, hot carcass weight; CCW, cold carcass weight.

Download Excel Table
Table 4. Wholesale cuts of the hair lambs supplemented with FA and CTL
Item FA (ppm)1) CTL (%)2) SEM p-value
0 300 0 1 FA CTL FA × CTL
Neck (%) 8.66 7.60 8.31 7.94 0.64 0.134 0.592 0.957
Shoulder (%) 29.02 30.26 29.51 29.77 0.96 0.238 0.800 0.374
Loin (%) 7.99 8.01 8.30 7.70 0.60 0.974 0.358 0.679
Ribs and flank (%) 14.08 13.79 14.13 13.74 0.72 0.702 0.613 0.574
Plain loin (%) 11.35 11.17 12.10 10.42 1.57 0.913 0.321 0.424
Leg (%) 29.21 28.44 27.33 30.31 1.67 0.668 0.109 0.524

Lambs fed 0 or 300 ppm of ferulic acid through the 40-d feeding trial.

Lambs fed 0 or 1% of clinoptilolite through the 40-d feeding.

FA, ferulic acid; CTL, clinoptilolite.

Download Excel Table
Meat quality evaluation

Neither the FA × CTL interaction nor the additives themselves had any impact on pH 24 h, WHC, cooking loss, or WBSF (p > 0.05) (Table 5). The final pH was unaltered (p > 0.05) by FA or CTL supplementation, while their interaction (FAZ treatment) tended (p = 0.095) to decrease the final FAZ sample pH (pH = 5.67; unpublished raw data from the study). Regarding color variables, dietary FA did not have any effect on the L*, a*, b*, or C* values (p > 0.05; Table 5). On the other hand, dietary CTL improved (p < 0.05) the L*, a*, and C* values, whereas it tended to increase yellowness (b*; p = 0.070). The hue angle (H*) remained unaltered for all treatments (p > 0.05).

Table 5. Effect of FA and CTL on meat quality of the hair lambs
Item FA (ppm)1) CTL (%)2) SEM p-value
0 300 0 1 FA CTL FA × CTL
pH 24 h 5.50 5.49 5.53 5.45 0.08 0.923 0.391 0.317
Final pH 6.01 5.76 5.86 5.92 0.142 0.116 0.686 0.095
WHC (%) 79.52 81.41 80.79 80.14 1.30 0.183 0.639 0.612
Color variables
 L* 38.63 38.09 37.43 39.28 0.804 0.527 0.041 0.703
 a* 17.83 17.54 16.91 18.46 0.642 0.668 0.034 0.178
 b* 8.83 8.34 7.93 9.24 0.649 0.482 0.070 0.167
 Chroma (C*) 19.91 19.45 18.70 20.66 0.837 0.604 0.038 0.160
 Hue angle (H*) 26.22 25.09 24.93 26.39 1.07 0.328 0.209 0.265
 Cooking loss (%) 15.97 17.21 17.30 15.87 1.98 0.558 0.500 0.456
 WBSF (kg) 7.36 6.80 7.60 6.56 0.850 0.539 0.258 0.623

Lambs fed 0 or 300 ppm of ferulic acid through the 40-d feeding trial.

Lambs fed 0 or 1% of clinoptilolite through the 40-d feeding.

FA, ferulic acid; CTL, clinoptilolite; WHC, water-holding capacity; WBSF, Warner-Bratzler shear force.

Download Excel Table
Chemical composition and fatty acid profile

The chemical composition of the lamb meat is shown in Table 6. In general, the chemical composition of the lamb meat was not influenced (p > 0.05) by the additives themselves or by their interaction in terms of moisture, ash, fat, or protein contents. Table 7 presents the influence of dietary supplementation with FA and CTL on the fatty acid profile of the intramuscular fat of hair lambs. Palmitic (C16:0), stearic (C18:0), and oleic (C18:1ɷ9c) acids represent the greatest proportion of the fatty acid profile of the lambs’ LT muscle in all treatments. FA supplementation did not have any effect (p > 0.05) on the fatty acid profile of the lamb meat, whereas the CTL and FA x CTL interaction modified the content of some fatty acids (Table 7). CTL increased (p < 0.05) the percentages of C10:0, C14:1, and C15:0 (18, 40, and 22%, respectively) in comparison to animals not supplemented; meanwhile, C14:0 decreased (p > 0.05) 13.30% by CTL supplementation. Additionally, CTL tended to reduce C16:0 (p = 0.068; 5.4%) and C20:1ɷ9 (p = 0.060; 26.3%). Regarding C18:0, C18:2ɷ6t, and C20:3ɷ6, the FA x CTL interaction increased (p < 0.05) the content of the aforementioned fatty acids while it tended to increase C17:0 (p = 0.098) and C20:3ɷ6 (p = 0.097). No FA × CTL interaction or FA main effect (p > 0.05) was presented for any of the sums of fatty acids and nutritional indices of lamb meat (Table 8). However, while CTL tended to increase the sum of PUFAs (p = 0.053; 12.04%) and n-6 (p = 0.066; 11.87%) and the PUFA/SFA ratio (p = 0.070; 21.05%), the other sums and nutritional indices remained unaltered (p > 0.05).

Table 6. Chemical composition of meat from hair lambs supplemented with FA and CTL
Chemical composition (%) FA (ppm)1) CTL (%)2) SEM p-value
0 300 0 1 FA CTL FA × CTL
Moisture 72.11 73.05 73.32 71.84 0.928 0.350 0.143 0.697
Ash 0.98 0.98 0.98 0.98 0.031 0.893 0.793 0.102
Fat 4.19 5.39 4.64 4.94 1.109 0.310 0.794 0.194
Protein 21.74 20.64 21.30 21.08 0.768 0.182 0.791 0.219

Lambs fed 0 or 300 ppm of ferulic acid through the 40-d feeding trial.

Lambs fed 0 or 1% of clinoptilolite through the 40-d feeding.

FA, ferulic acid; CTL, clinoptilolite.

Download Excel Table
Table 7. Fatty acids profile of the longissimus thoracis muscle from hair lambs supplemented with FA and CTL
Fatty acid (%) FA (ppm)1) CTL (%)2) SEM p-value
0 300 0 1 FA CTL FA × CTL
C10:0 0.11 0.13 0.11 0.13 0.007 0.108 0.038 0.266
C12:0 0.08 0.09 0.09 0.08 0.008 0.558 0.150 0.203
C14:0 2.01 2.06 2.18 1.89 0.114 0.728 0.035 0.138
C14:1 0.76 0.72 0.62 0.868 0.074 0.649 0.008 0.672
C15:0 0.61 0.58 0.54 0.658 0.043 0.541 0.039 0.672
C15:1 1.38 1.39 1.36 1.408 0.189 0.963 0.846 0.462
C16:0 22.53 21.91 22.84 21.60 0.591 0.338 0.068 0.647
C16:1 2.65 2.35 2.45 2.553 0.157 0.122 0.606 0.278
C17:0 1.56 1.55 1.56 1.555 0.080 0.947 0.911 0.098
C17:1 1.00 1.05 0.94 1.107 0.098 0.642 0.152 0.152
C18:0 15.88 15.17 15.78 15.27 0.497 0.211 0.365 0.019
C18:1ɷ9c 42.56 42.20 42.37 42.4 0.602 0.586 0.964 0.939
C18:2ɷ6t 0.48 0.44 0.45 0.479 0.028 0.190 0.319 0.019
C18:2ɷ6c 5.68 5.65 5.33 6.01 0.378 0.946 0.116 0.680
C18:3ɷ3 0.38 0.40 0.39 0.39 0.034 0.616 0.963 0.043
C20:1ɷ9 0.15 0.18 0.19 0.14 0.029 0.301 0.060 0.288
C20:2 0.32 0.36 0.34 0.34 0.047 0.466 0.944 0.229
C20:3ɷ6 2.02 1.89 1.89 2.01 0.132 0.365 0.418 0.097
C20:5ɷ3 0.27 0.26 0.28 0.25 0.044 0.814 0.575 0.148
C24:0 0.33 0.37 0.37 0.34 0.035 0.350 0.475 0.102

Lambs fed 0 or 300 ppm of ferulic acid through the 40-d feeding trial.

Lambs fed 0 or 1% of clinoptilolite through the 40-d feeding.

FA, ferulic acid; CTL, clinoptilolite.

Download Excel Table
Table 8. Sums of fatty acids and nutritional indices value of the longissimus thoracis muscle from hair lambs supplemented with FA and CTL
Item FA (ppm)1) CTL (%)2) SEM p-value
0 300 0 1 FA CTL FA × CTL
∑ SFA 43.2 44.06 44.71 42.55 1.40 0.556 0.155 0.908
∑ MUFA 47.34 47.72 46.86 48.20 1.13 0.760 0.286 0.510
∑ PUFA 9.42 9.07 8.72 9.77 0.48 0.494 0.053 0.311
MUFA / SFA 1.06 1.07 1.05 1.09 0.059 0.948 0.501 0.613
PUFA / SFA 0.21 0.21 0.19 0.23 0.017 0.788 0.070 0.114
∑ n-3 0.65 0.66 0.65 0.65 0.068 0.871 0.983 0.686
∑ n-6 8.41 8.02 7.75 8.67 0.438 0.418 0.066 0.484
n-6 / n-3 13.70 12.29 12.69 13.30 1.09 0.218 0.581 0.869

Lambs fed 0 or 300 ppm of ferulic acid through the 40-d feeding trial.

Lambs fed 0 or 1% of clinoptilolite through the 40-d feeding.

FA, ferulic acid; CTL, clinoptilolite; SFA, saturated fatty acid; MUFA, monounsaturated fatty acid; PUFA, polyunsaturated fatty acid.

Download Excel Table

DISCUSSION

The findings of the present study did not show any additive or synergistic effect in the supplemented animals that could support our hypothesis; therefore, the discussion focuses on the individual impact of each supplement.

Feedlot performance

The mechanism of action of FA has not been fully elucidated. Based on previous studies with this compound, it has proposed different mechanisms of action involved in the growth-promoting effect of FA: 1) FA could enhance animal performance by reducing their oxidative stress [28]. 2) FA has a similar action mechanism to beta-agonists (βAAs) due to the similarities of their molecular structures [29,30]. 3) FA acts as a modulator of ruminal fermentation [12,31].

In our current study, we have no clear information to explain why FA supplementation did not improve feedlot performance traits, carcass characteristics, or wholesale cuts of the finished lambs. In agreement with our results, other studies reported that dietary FA administration did not affect the growth performance of hair lambs [8-10,32]. Contrary to our results, Peña-Torres et al. [30] observed that feeding FA (300 ppm and 600 ppm) enhanced the ADG of lambs; however, in that study, the lambs had a lower body weight and age than the animals in our study. This growth-promoting effect reported in the last study can be related to the high growth rate of the lambs at younger ages, which is usually the period when lambs have continuous growth without reaching their inflection point in the growth curve [33]. Nevertheless, an anabolic effect has been shown in other species, such as pigs and beef cattle [7,34]. Further research on beta-adrenergic receptor (β-AR; subtypes β1, β2 and β3) gene expression or receptor binding affinity evaluation is needed to elucidate these inconsistencies between species.

Based on the aforementioned β–AR gene expression, Valenzuela-Grijalva et al. [29] studied the change in pigs’ skeletal muscle mRNA abundance after FA supplementation. β2-AR mRNA expression was increased by FA intake. Baxa et al. [35] reported a similar effect in finishing steers by zilpaterol hydrochloride supplementation. It is worth mentioning that the β2-AR subtype is predominant in bovine and porcine skeletal muscle [36]. On the other hand, Ekpe et al. [37] found an increase in the receptor density of β1-AR in sheep under feed restriction and observed a trend toward cold stress. Previous studies have reported a low plasma concentration of endogenous catecholamines in lambs under thermoneutral conditions. Thus, there might be a lower activation of skeletal muscle β-receptors, which results in less receptor binding affinity [37,38]. These observations suggest that in pigs and beef cattle, FA stimulates β2-AR to promote a growth effect, while in lambs, FA behaves differently.

Based on the above, the lack of effects on the feedlot performance of lambs supplemented with FA can be explained. However, the lambs in our study significantly decreased their feed intake without affecting their performance, which could represent an advantage in meat production development because feeding is an element that elevates production costs. In contrast with our results, other studies with lambs found no effect of FA or feruloyl oligosaccharides on feed intake [8-10,32]. Additionally, published reports have studied the effect of supplementing lambs with different phytochemicals or plant extracts; their results have shown that feeding these compounds either increased or did not affect the feed intake of fattening lambs [39]. In our study, a decrease in feed intake was only present in the second period (days 21 to 40), and this reduction did not affect the lambs’ weight gain. In this regard, prolonged feeding of FA could positively modify the ruminal environment and, consequently, some ruminal fermentation patterns, nutrient digestion, or energy utilization [12,31].

On the other hand, dietary supplementation of fattening lambs with CTL tended to increase feed intake in each experimental period. Similarly, several studies have reported that CTL supplementation increased the daily feed intake of livestock [13,40,41]. Otherwise, the lambs’ weight gain in our experiment was not affected by CTL supplementation, while in other studies, dietary supplementation with CTL has been proven to increase weight gain in lambs [18,40,41]. This lack of effect on the weight gain of the lambs might be due to the limited sample size used in our study. On the other hand, Pond et al. [42] mentioned that a CTL advantage on lamb growth performance appeared when diets contained an intact protein source and a high protein level. According to the previous study, it is suggested that high protein intake elevates plasma urea-nitrogen and ruminal ammonium (NH4+) concentrations [42]. Since CTL has a high NH4+ binding capacity, NH4+ ions are immediately exchanged for the compensating cation of the zeolite and held there until they are released when saliva enters the rumen during rumination. It is possible that the dosed release of NH4+ ions enhanced ruminal bacteria growth, which in turn could improve ruminal fermentation patterns and microbial protein production. Moreover, Abdelrahman et al. [41] observed that lambs treated with 1% zeolite presented a similar performance to the lambs of the control group, while lambs fed with 2% zeolite showed a better growth performance. In the present work, the failure of CTL to affect weight gain in hair-breed lambs might be due to either an inappropriately low dose of CTL (chosen in this study) or balanced protein intake.

Carcass characteristics and wholesale cuts yield

FA supplementation did not affect either the carcass characteristics or the wholesale cuts. The only change shown in these traits was a reduction in the conformation of the FA carcasses. The lack of effects of FA in our results is in accordance with [9,10,30] findings. In contrast, Peña-Torres et al. [6] found that by including 250 ppm FA in the diet of heifers, dressing weight and loin area were improved. In addition, previous studies have shown that phytochemicals and plant extracts did not alter carcass characteristics in fattening lambs [39]. Although our values were within the reference ranges, the lack of an effect on the carcass traits and wholesale cuts suggests that FA supplementation does not have anabolic stimulation in hair-breed lambs, as shown in younger lambs and other species [7,9,29,30]. Since the results of carcass characteristics and wholesale cuts have not been consistent throughout studies, these inconsistencies may be due to the growth effect shown by FA is associated with the breed, sex, species, environmental conditions, or the dose used [43].

Regarding CTL addition, there was no evidence of remarkable effects on carcass characteristics and wholesale cuts. Previous reports indicate that CTL did not affect any carcass characteristics of hair-breed lambs [15,40,43]. Since the CTL supplementation did not improve lambs’ muscular development in our study, consequently, no changes in carcass characteristics were expected. Deligiannis et al. [40] reported that lambs supplemented with 3% of CTL showed a better weight gain with no change on carcass traits. Similarly, other studies have reported that CTL supplementation enhances weight gain (carcass characteristics were not evaluated) [17,41,42]. The effects of CTL supplementation in lambs in other studies, could be related to the ability of CTL to modify ruminal fermentation due to its cation exchange property, resulting in better energy efficiency by improving the acetate:propionate ratio or by stimulating some populations of ruminal bacteria [18,44,45].

Meat quality

In the present study, FA supplementation did not affect the meat quality of hair lambs. These parameters (pH, WHC, cooking loss, color, and WBSF) were maintained within the reported values for lambs [9,46-48]. The lack of effects on color parameters between groups is consistent with previous reports with FA supplementation [6-9,48]. It has been reported that phytochemicals that exhibited antioxidant properties could delay metmyoglobin formation by reducing myoglobin and lipid oxidation, improving meat color stability [5,49]; however, FA did not show any antioxidant properties in our study. Phytochemicals (such as FA) are biotransformed by methylation, glucuronidation, and sulphation into conjugated metabolites because the body recognizes phytochemicals as xenobiotics. Therefore, after intake, concentrations of phytochemicals remain relatively low. Thus, these metabolites can partially reach the target tissues. Besides, they are rapidly excreted via urine and bile [28], limiting their tissue accumulation.

To our knowledge, this is the first report to evaluate meat quality in hair lambs supplemented with CTL. Meat quality characteristics (WHC, pH, WBSF, and cooking loss) were not influenced by CTL supplementation. The only change shown in these traits was an increase in some color variables. Information about the influence of CTL on meat color parameters is limited. In the current study, CTL supplementation improved L*, a*, and C* in the meat of lambs. In contrast, Hcini et al. [13] reported no change in the color of breast meat of turkeys supplemented with 1% and 2% CTL. No antioxidant status of lambs was measured in this work; however, the improvement observed in color parameters of lamb meat could be attributed to the antioxidant activity of CTL [49]. The antioxidant capacity of CTL is associated with the scavenging of reactive oxygen species (ROS) and transition metals or to the increased activity of endogenous antioxidant enzymes (glutathione peroxidase, superoxide dismutase, catalase, and nitric oxide synthase) by offering cofactors such as trace elements selenium, copper, zinc, and manganese within the CTL structure [13,50]. As mentioned above, we hypothesize that the microelements contained in CTL could reach muscle tissue and perform its antioxidant activity. However, future research is needed to demonstrate the fate and effect of CTL on meat quality of lambs.

Chemical composition and fatty acid profile

The addition of FA or CTL to the lamb diet did not negatively affect the chemical composition of lamb meat. Moisture, ash, fat, and protein were maintained within the range reported by other authors [46,47] for hair lambs. Since the fat content was slightly higher than that reported in other studies, we believe that this result may be attributed to the older lambs used in this study. Mature animals deposit around 85% of their energy as fat [51].

In the current study, FA supplementation did not change the fatty acid profile, the overall fatty acids, or the nutritional indices of hair lambs, whereas CTL and FA × CTL positively modified the fatty acid profile. In agreement with our findings, Mallek et al. [52] reported a decrease in myristic (C14:0), palmitic (C16:0), and stearic (C18:0) acids in broilers’ fatty acid profile by zeolite supplementation as well as an increase in linolenic acid (C18:3ɷ3). Additionally, a study reported a reduction in DHA (C22:6 n-3) and eicosenoic acid (C20:1 n-9) in lambs’ fatty acid profile by monensin sodium supplementation (33 mg/kg of Rumensin 200® for 70 d) [53]. The potential pathways by which CTL or FA × CTL modify lambs’ fatty acid profile is unclear, but an enhancement of the fatty acid desaturase activity [13,52] or changes in the rumen microbial population [53,54], could be involved.

Fatty acids desaturases such as Δ9-desaturase introduce a double bond across carbons of a SFAs carbon chain. A study reported that a higher desaturase product in the muscle of supplemented kids (Terminalia chebula extract 2 and 6 mg/mL of rumen volume) is due to the action of the Δ9-desaturase enzyme, suggesting that the desaturase activity was influenced by the presence of phenolic compounds [55]. Also, Δ9-desaturase activity could be regulated by some elements in CTL [13-50,52]. Concerning the second pathway proposed, CTL ion-exchange activity may modify the ruminal environment, resulting in changes in the rumen microbial population, especially in cellulolytic bacteria [44]. Cellulolytic bacteria such as Ruminococcus albus y Butyrivibrio sp. are placed in the first group of rumen bacteria which hydrogenate unsaturated fatty acids. Butyrivibrio fibrisolvens have been used to explore the multiple biohydrogenation (BH) pathways. These bacteria can hydrogenate linoleic and alfa-linolenic acids to a conjugated form as the final product (without forming stearic acid as the end products) [54]. Based on the aforementioned, our results could be caused by a regulation of Δ9-desaturase activity or by a slight inhibition of the BH of unsaturated fatty acids [53,54], which may increase the total PUFA, n-6, and n-3 contents and enhance their nutritional ratios. Nevertheless, further studies are needed to show whether changes in fatty acid profile were modified via regulation of Δ9-desaturase activity or by changes in the ruminal microbial population.

In addition, although the ∑ PUFA, ∑ n-6, and PUFA/SFA ratios in our results did not reach significance, these values tended to improve, which may lead to healthier meat from a nutritional viewpoint [56]. Recently, there has been an interest in improving the nutritional indices of meat, as high values of PUFA/SFA and n-6/n-3 ratios represent a risk factor for cancer and coronary heart disease. The recommended ratio of n-6/n-3 is less than 4, and PUFA/SFA should be above 0.4 [56]. In general, in this study, the n-6/n-3 ratio was high for all treatments, which may have occurred due to the low content of omega-3 fatty acids found in our results. In the current work, DHA (C22:6 n-3) was not detectable, and eicosapentaenoic acid (EPA; C20:5 n-3) was found at a low concentration in lamb meat [47-53,57] in all treatments, which could explain the undesirably high n-6/n-3 ratio found in this work.

CONCLUSION

Feeding FA and CTL did not have any combined effect on the growth performance, carcass characteristics, wholesale cut yields, or meat quality of hair lambs. This study provides information about how adding FA reduces daily feed intake without compromising weight gain, which could represent an advantage for lowering feeding costs.

Additionally, this experiment provides new evidence that CTL supplementation improves some meat quality traits and the fatty acid profile of hair lambs. Nevertheless, further studies in ruminants, including a high CTL dose, are needed in order to understand the conditions of CTL beneficial effects on feedlot performance traits.

Factors such as dosage, age, diet composition, and environmental conditions need more investigation to elucidate the beneficial effects of FA and CTL supplementation on hair lambs. Besides, further research is necessary to investigate the effects of FA and CTL supplementation on the ruminal fermentation parameters and microbial populations of the rumen of hair-breed lambs to complement the findings in this work.

Competing interests

No potential conflict of interest relevant to this article was reported.

Funding sources

This work was supported by the Centro de Investigación en Alimentación y Desarrollo A. C. and Carnes Tapeke´nos S.A. de C.V. in Mexico.

Acknowledgements

We appreciate the donation of supplies by Roberto Laguna, Felizardo Leon and Geovanni Leyva. We thank the support provided in the laboratory by M.C. Libertad Zamorano, Julio González, Francisco Alday, and Laura Flores. Also, we acknowledge Leonardo Lozano, Sebastian Ibarra, and Carnes Tapeke´nos agricultural operators for their facilities.

Availability of data and material

Upon reasonable request, the datasets of this study can be available from the corresponding author.

Authors’ contributions

Conceptualization: González-Ríos H.

Data curation: Tánori-Lozano A, Pinelli-Saavedra A, Valenzuela-Melendres M.

Formal analysis: Tánori-Lozano A, Quintana-Romandía AI, Dávila-Ramírez JL.

Methodology: Tánori-Lozano A, Quintana-Romandía AI, Montalvo-Corral M, Valenzuela-Melendres M, Islava-Lagarda TY, González-Ríos H.

Investigation: Tánori-Lozano A, Quintana-Romandía AI, Pinelli-Saavedra A, Islava-Lagarda TY, González-Ríos H.

Writing - original draft: Tánori-Lozano A, Dávila-Ramírez JL, González-Ríos H.

Writing - review & editing: Tánori-Lozano A, Quintana-Romandía AI, Montalvo-Corral M, Pinelli-Saavedra A, Valenzuela-Melendres M, Dávila-Ramírez JL, Islava-Lagarda TY, González-Ríos H.

Ethics approval and consent to participate

The study was conducted according to the local official guidelines for animal care in Mexico, and approved by the Research Ethics Committee of the Centro de Investigación en Alimentación y Desarrollo A.C. (CEI/002-2/2021).

REFERENCES

1.

Heitzman RJ. The efficacy and mechanism of action of anabolic agents as growth promoters in farm animals. J Steroid Biochem. 1979; 11:927-30

2.

Gonzalez Ronquillo M, Angeles Hernandez JC. Antibiotic and synthetic growth promoters in animal diets: review of impact and analytical methods. Food Control. 2017; 72:255-67

3.

Dávila-Ramírez JL, Avendaño-Reyes L, Macías-Cruz U, Torrentera-Olivera NG, Zamorano-García L, Peña-Ramos A, et al. Effects of zilpaterol hydrochloride and soybean oil supplementation on physicochemical and sensory characteristics of meat from hair lambs. Small Rumin Res. 2013; 114:253-7

4.

Lillehoj H, Liu Y, Calsamiglia S, Fernandez-Miyakawa ME, Chi F, Cravens RL, et al. Phytochemicals as antibiotic alternatives to promote growth and enhance host health. Vet Res. 2018; 49:76

5.

González-Ríos H, Dávila-Ramírez JL, Peña-Ramos EA, Valenzuela-Melendres M, Zamorano-García L, Islava-Lagarda TY, et al. Dietary supplementation of ferulic acid to steers under commercial feedlot feeding conditions improves meat quality and shelf life. Anim Feed Sci Technol. 2016; 222:111-21

6.

Peña-Torres EF, Dávila-Ramírez JL, Peña-Ramos EA, Valenzuela-Melendres M, Pinelli-Saavedra A, Avendaño-Reyes L, et al. Effects of dietary ferulic acid on growth performance, carcass traits and meat quality of heifers. J Sci Food Agric. 2021; 101:548-54

7.

González Ríos H, Lozano DAG, Mir AB. Ferulic acid as feed supplement in beef cattle to promote animal growth and improve the meat quality of the carcass and the meat. United States patent US 20130041036A1 2013 Fed; 14

8.

Wang Y, Wang W, Wang R, Meng Z, Duan Y, An X, et al. Dietary supplementation of ferulic acid improves performance and alleviates oxidative stress of lambs in a cold environment. Can J Anim Sci. 2019; 99:705-12

9.

Valadez-García KM, Avendaño-Reyes L, Díaz-Molina R, Mellado M, Meza-Herrera CA, Correa-Calderón A, et al. Free ferulic acid supplementation of heat-stressed hair ewe lambs: oxidative status, feedlot performance, carcass traits and meat quality. Meat Sci. 2021; 173:108395

10.

Macías-Cruz U, Perard S, Vicente R, Álvarez FD, Torrentera-Olivera NG, González-Ríos H, et al. Effects of free ferulic acid on productive performance, blood metabolites, and carcass characteristics of feedlot finishing ewe lambs. J Anim Sci. 2014; 92:5762-8

11.

Soberon MA, Cherney DJR, Cherney JH. Free ferulic acid uptake in ram lambs. J Anim Sci. 2012; 90:1885-91

12.

Bodas R, Prieto N, García-González R, Andrés S, Giráldez FJ, López S. Manipulation of rumen fermentation and methane production with plant secondary metabolites. Anim Feed Sci Technol. 2012; 176:78-93

13.

Hcini E, Slima AB, Kallel I, Zormati S, Traore AI, Gdoura R. Does supplemental zeolite (clinoptilolite) affect growth performance, meat texture, oxidative stress and production of polyunsaturated fatty acid of Turkey poults?. Lipids Health Dis. 2018; 17:177

14.

Valpotić H, Gračner D, Turk R, Đuričić D, Vince S, Folnožić I, et al. Zeolite clinoptilolite nanoporous feed additive for animals of veterinary importance: potentials and limitations. Period Biol. 2017; 119:159-72

15.

Coronel-Burgos F, Plascencia A, Castro-Pérez BI, Contreras-Pérez G, Barreras A, Estrada-Angulo A. Influencia de la sustitución parcial del maíz y de la pasta de soja por zeolita en ovinos en etapa de finalización: características de la canal, composición tisular y masa visceral. Arch Zootec. 2017; 66:223-8

16.

Estrada-Angulo A, Coronel-Burgos F, Castro Pérez BI, López Soto MA, Barreras A, Angulo-Montoya C, et al. Efecto de la inclusión de zeolita (clinoptilolita) en ovinos en etapa de finalización: respuesta productiva y energética de la dieta. Arch Zootec. 2017; 66:381-6

17.

Roque-Jiménez JA, Pinos-Rodríguez JM, Rojo-Rub R, Mendoza GD, Vazquez A, Cayetano De Jesus JA, et al. Effect of natural zeolite on live weight changes, ruminal fermentation and nitrogen metabolism of ewe lambs. S Afr J Anim Sci. 2018; 48:1148-55

18.

Toprak NN, Yılmaz A, Öztürk E, Yigit O, Cedden F. Effect of micronized zeolite addition to lamb concentrate feeds on growth performance and some blood chemistry and metabolites. S Afr J Anim Sci. 2016; 46:313-20

19.

NRC [National Research Council], Committee on Nutrient Requirements of Small Ruminants. Nutrient requirements of small ruminants: sheep, goats, cervids, and new world camelids. Washington, DC: The National Academies Press. 2007

20.

AMSA [American Meat Science Association]. Meat evaluation handbook. Savoy, IL: American Meat Science Association. 2001

21.

Parés-Casanova PM. Morphometric dimensions allow differentiation of lamb carcasses for some breeds. Egypt J Sheep Goats Sci. 2013; 8:167-70

22.

Avendaño-Reyes L, Macías-Cruz U, Álvarez-Valenzuela FD, Águila-Tepato E, Torrentera-Olivera NG, Soto-Navarro SA. Effects of zilpaterol hydrochloride on growth performance, carcass characteristics, and wholesale cut yield of hair-breed ewe lambs consuming feedlot diets under moderate environmental conditions. J Anim Sci. 2011; 89:4188-94

23.

Sutton DS, Ellis M, Lan Y, McKeith FK, Wilson ER. Influence of slaughter weight and stress gene genotype on the water-holding capacity and protein gel characteristics of three porcine muscles. Meat Sci. 1997; 46:173-80

24.

Cassens RG, Demeyer D, Eikelenboom G, Honikel KO, Johansson G, Nielsen T, et al. Recommendation of reference method for assessment of meat color.In In: Proceedings of 41st Annual International Congress of Meat Science and Technologyvol2 1995; , San Antonio, TXp p. 410-1

25.

AOAC [Association of Official Analytical Chemists] International. Official methods of analysis of AOAC International. 17th ed Gaithersburg, MD: AOAC International. 2000

26.

Bligh EG, Dyer WJ. A rapid method of total lipid extraction and purification. Can J Biochem Physiol. 1959; 37:911-7

27.

Li Y, Watkins BA. Analysis of fatty acids in food lipids. Curr Protoc Food Anal Chem. 2001; (1):D1. 2.1-15

28.

Gessner DK, Ringseis R, Eder K. Potential of plant polyphenols to combat oxidative stress and inflammatory processes in farm animals. J Anim Physiol Anim Nutr. 2017; 101:605-28

29.

Valenzuela-Grijalva N, Jiménez-Estrada I, Mariscal-Tovar S, López-García K, Pinelli-Saavedra A, Peña-Ramos EA, et al. Effects of ferulic acid supplementation on growth performance, carcass traits and histochemical characteristics of muscle fibers in finishing pigs. Animals. 2021; 11:2455

30.

Peña-Torres EF, Castillo-Salas C, Jiménez-Estrada I, Muhlia-Almazán A, Peña-Ramos EA, Pinelli-Saavedra A, et al. Growth performance, carcass traits, muscle fiber characteristics and skeletal muscle mRNA abundance in hair lambs supplemented with ferulic acid. J Anim Sci Technol. 2022; 64:52-69

31.

Patra AK, Saxena J. Dietary phytochemicals as rumen modifiers: a review of the effects on microbial populations. Antonie van Leeuwenhoek. 2009; 96:363-75

32.

Wang Y, Meng Z, Guo J, Wang W, Duan Y, Hao X, et al. Effect of wheat bran feruloyl oligosaccharides on the performance, blood metabolites, antioxidant status and rumen fermentation of lambs. Small Rumin Res. 2019; 175:65-71

33.

Domínguez-Viveros J, Canul-Santos E, Rodríguez-Almeida FA, Burrola-Barraza ME, Ortega-Gutiérrez JÁ, Castillo-Rangel F. Defining growth curves with nonlinear models in seven sheep breeds in Mexico. Rev Mex Cienc Pecu. 2019; 10:664-75

34.

Herrera RH, Castillo MLA, Torres AJA. Methods to accelerate muscle development, decrease fat deposits, and enhance feeding efficiency in pigs. United States Patent US 20110046224A1 2011 Feb; 24

35.

Baxa TJ, Hutcheson JP, Miller MF, Brooks JC, Nichols WT, Streeter MN, et al. Additive effects of a steroidal implant and zilpaterol hydrochloride on feedlot performance, carcass characteristics, and skeletal muscle messenger ribonucleic acid abundance in finishing steers. J Anim Sci. 2010; 88:330-7

36.

Sato S, Shirato K, Tachiyashiki K, Imaizumi K. Muscle plasticity and β2-adrenergic receptors: adaptive responses of β2-adrenergic receptor expression to muscle hypertrophy and atrophy. J Biomed Biotechnol. 2011; 2011:729598

37.

Ekpe ED, Moibi JA, Christopherson RJ. Beta-adrenergic receptors in skeletal muscles of ruminants: effects of temperature and feed intake. Can J Anim Sci. 2000; 80:79-86

38.

Thompson JR, Christopherson RJ, Hammond VA, Hills GA. Effects of acute cold exposure on plasma concentrations of noradrenaline and adrenaline in sheep. Can J Anim Sci. 1978; 58:23-8

39.

Yagoubi Y, Smeti S, Ben Saïd S, Srihi H, Mekki I, Mahouachi M, et al. Carcass traits and meat quality of fat-tailed lambs fed rosemary residues as a part of concentrate. Animals. 2021; 11:655

40.

Deligiannis K, Lainas T, Arsenos G, Papadopoulos E, Fortomaris P, Kufidis D, et al. The effect of feeding clinoptilolite on food intake and performance of growing lambs infected or not with gastrointestinal nematodes. Livest Prod Sci. 2005; 96:195-203

41.

Abdelrahman MM, Alhidary I, Adeniji YA, Alobre MM, Albaadani H, Aljumaah R. Manipulating phosphorus, calcium, and magnesium utilization by growing lambs using natural zeolite (clinoptilolite). Sustainability. 2021; 13:1539

42.

Pond WG, Laurent SM, Orloff HD. Effect of dietary clinoptilolite or zeolite Na-A on body weight gain and feed ulilization of growing lambs fed urea or intact protein as a nitrogen supplement. Zeolites. 1984; 4:127-32

43.

Valadez-García KM, Avendaño-Reyes L, Meza-Herrera CA, Mellado M, Díaz-Molina R, González-Ríos H, et al. Ferulic acid in animal feeding: mechanisms of action, productive benefits, and future perspectives in meat production. Food Biosci. 2021; 43:101247

44.

Goodarzi M, Nanekarani S. The effects of calcic and potassic clinoptilolite on ruminal parameters in lori breed sheep. APCBEE Procedia. 2012; 4:140-5

45.

Urías-Estrada JD, López-Soto MA, Barreras A, Aguilar-Hernández JA, González-Vizcarra VM, Estrada-Angulo A, et al. Influence of zeolite (clinoptilolite) supplementation on characteristics of digestion and ruminal fermentation of steers fed a steam-flaked corn-based finishing diet. Anim Prod Sci. 2017; 58:1239-45

46.

López-Baca MÁ, Avendaño-Reyes L, Macías-Cruz U, Muhlia-Almazán A, Valenzuela-Melendres M, Peña-Ramos EA, et al. Muscle fiber morphometry and physicochemical characteristics of the Longissimus thoracis muscle of hair male lambs fed zilpaterol hydrochloride and implanted with steroids. Meat Sci. 2021; 177:108490

47.

Dávila-Ramírez JL, Avendaño-Reyes L, Macías-Cruz U, Peña-Ramos EA, Islava-Lagarda TY, Zamorano-García L, et al. Fatty acid composition and physicochemical and sensory characteristics of meat from ewe lambs supplemented with zilpaterol hydrochloride and soybean oil. Anim Prod Sci. 2016; 57:767-77

48.

Peña E, Gonzalez H, Rosales F, Jiménez I, Valenzuela M, Peña A, et al. PSII-21 fiber type characterization and meat quality of hair lambs supplemented with ferulic acid. J Anim Sci. 2018; 96:75

49.

Simitzis PE, Deligeorgis SG, Bizelis JA, Dardamani A, Theodosiou I, Fegeros K. Effect of dietary oregano oil supplementation on lamb meat characteristics. Meat Sci. 2008; 79:217-23

50.

Kraljević Pavelić S, Simović Medica J, Gumbarević D, Filošević A, Pržulj N, Pavelić K. Critical review on zeolite clinoptilolite safety and medical applications in vivo. Front Pharmacol. 2018; 9:1350

51.

Fiems LO. Effect of beta-adrenergic agonists in animal production and their mode of action. Ann Zootech. 1987; 36:271-90

52.

Mallek Z, Fendri I, Khannous L, Hassena AB, Traore AI, Ayadi MA, et al. Effect of zeolite (clinoptilolite) as feed additive in Tunisian broilers on the total flora, meat texture and the production of omega 3 polyunsaturated fatty acid. Lipids Health Dis. 2012; 11:35

53.

Garcia-Galicia IA, Arras-Acosta JA, Huerta-Jimenez M, Rentería-Monterrubio AL, Loya-Olguin JL, Carrillo-Lopez LM, et al. Natural oregano essential oil may replace antibiotics in lamb diets: effects on meat quality. Antibiotics. 2020; 9:248

54.

Jenkins TC, Wallace RJ, Moate PJ, Mosley EE. Board-invited review: recent advances in biohydrogenation of unsaturated fatty acids within the rumen microbial ecosystem. J Anim Sci. 2008; 86:397-412

55.

Rana MS, Tyagi A, Hossain SA, Tyagi AK. Effect of tanniniferous Terminalia chebula extract on rumen biohydrogenation, ∆9-desaturase activity, CLA content and fatty acid composition in longissimus dorsi muscle of kids. Meat Sci. 2012; 90:558-63

56.

Great Britain Cardiovascular Review Group, Great Britain Department of Health. Nutritional aspects of cardiovascular disease: review Group Committee on Medical Aspects of Food Policy. London: H.M. Stationery Office. 1994

57.

de Evan T, Cabezas A, de la Fuente Vázquez J, Carro MD. Feeding agro-industrial by-products to light lambs: influence on meat characteristics, lipid oxidation, and fatty acid profile. Animals. 2020; 10:1572