ABSTRACT
This study evaluated the effects of whole flaxseed (WFS), fish oil (FO), and
different sources of Se, Zn, and Fe (inorganic, organic, and nano-source) on egg
production, quality, sensory attributes, and serum protein metabolites in laying
hens. A total of 144 hens were divided into six groups with six replicates of
four hens each. Hens were fed six diets as follows: 1) control diet; 2)
7.5%WFS+1.5%FO; 3) 7.5%WFS+1.5%FO+175 mg/kg vitamin E (VE); 4)
7.5%WFS+1.5%FO+175 mg/kg VE + inorganic sources of Se, Zn, and Fe (ISeZnFe); 5)
7.5%WFS+1.5%FO+175 mg/kg VE + organic sources of Se, Zn, and Fe (OSeZnFe); 6)
7.5%WFS+1.5%FO+175 mg/kg VE + nano-source of Se, Zn, and Fe (NSeZnFe) from 40–50
weeks of age. Laying hens fed 7.5% WFS, 1.5% FO, and different sources of trace
elements in their diets had no negative effects on laying rate, egg weight, egg
mass, feed intake, feed conversion ratio, body weight change, or survival rate
compared to that of hens fed the control diet. Dietary treatments did not
negatively affect the external and internal egg characteristics or egg sensory
attributes. Feeding 7.5%WFS+1.5%FO+VE+ISeZnFe positively influenced yolk color
in fresh eggs. Dietary treatments had a significant impact on egg nutritional
composition, with the highest levels of macronutrients found in eggs from hens
fed the 7.5%WFS+1.5%FO+VE+NSeZnFe treatment. The highest plasma globulin
concentrations were observed in hens fed organic and nano-source trace elements.
The same diets reduced plasma uric acid levels. Based on these findings, the
inclusion of organic or nano-source trace minerals in diets containing WFS and
FO positively affected egg quality and hen protein metabolites.
INTRODUCTION
Proteins are among the most important and expensive ingredients in commercial poultry
feed. Due to high costs and unpredictable supply, some protein sources are unable to
meet the rising demand, especially from poultry and aquaculture feed
manufacturers[1]. Recently, traditional
protein feed resources have been subject to a demand-supply mismatch, resulting in
increased feed costs and decreased sustainability of livestock production. In
addition, in regions where soybeans are not grown, there is a movement to replace
them with alternative sources of crop byproducts and vegetable protein for animal
nutrition[2].
Poultry nutrition has received special attention in poultry diets, with less
expensive feedstuffs receiving additional weight. In recent decades, there has been
greater focus on discovering ways to recycle feed manufacturing byproducts. Owing to
its high protein level and important amino acid content, soybean by-product, a
renewable resource, is considered a viable raw material for substituting or
supplementing animal, poultry, and fish meal[1]. Therefore, there is a growing interest in identifying alternatives to
soybean meal, such as other plant protein feeds and feed byproducts. Another purpose
of using crop co-products is to reduce feed costs and environmental pollution.
Moreover, these co-products are important sources of bioactive molecules with
significant advantages for human and animal health and the environment[3].
Marine products, including fish oil (FO) and plant products, which enhance the
nutritive value of poultry products (eggs and meat), are sources of omega-3
polyunsaturated fatty acids (n-3 PUFAs)[1,4,5]. Linseed or flaxseed (Linum usitatissimum L.) is an
important global annual crop valued as a feed additive and food because of its fiber
and oil content. Globally, linseed production of approximately 8,7000,000 tons in
2016 was priced at approximately $70.2[6].
Recently, there has been a growing interest in using flaxseed as a nutrient source
for both humans and animals because of the high levels of protein (20%–25%), oil
(35%–45%), metabolizable energy (3,800–3,960 kcal/kg), fiber (20%–25% including 10%
soluble fiber), and ash (3%–4%)[7,8]. Whole flaxseed (WFS) generally provides
approximately 40% oil, including 9%–10% of saturated fatty acids (mainly as palmitic
and stearic acids), 20% monounsaturated fatty acids (mainly oleic acid), and PUFAs
((mainly α-linolenic acid (ALA, 18:3, n-3))[9]. However, WFS contains many anti-nutritional factors (ANFs), such as
cyanogenic glycosides, tannins, phytic acid, trypsin inhibitors, anti-vitamin B6,
and non-starch polysaccharides, such as mucilage[10,11]. These ANFs impair
digestion and absorption and increase the viscosity of ingesta, which ultimately
limits the use of WFS in poultry nutrition[12]. Therefore, various techniques, including physicochemical and biological
methods, have been used to reduce ANFs[1,12,13].
Several studies have shown that WFS and its products may be used in poultry
production to enhance egg and meat quality[14]. Flaxseed, flaxseed meal, and flaxseed oil are the primary sources of
ALA in broiler chickens, laying hens[6], and
Japanese quail[15]. n-3
enriched eggs and poultry meat are successful strategies to reduce the incidence of
lifestyle diseases and enhance the human health index[16]. WFS (10%–20%) in the diets of laying hens increased ALA
levels in egg yolk by 10–20 fold[2].
Furthermore, Scheideler et al.[17] have
observed that 15% WFS in the diet increases the yolk ALA content by up to 7.1 g/100
g of fatty acids compared to that of a control diet. However, there is inconsistency
in the recommended WFS levels in chicken rations, including 2%–15% for
broilers[18,19] and up to 10% for laying hens[2,14]. Leeson et
al.[20] have observed that 20% ground
flaxseed in the diet has a negative effect on laying performance, compared to that
of zero and 10% treatments. Nam et al.[21]
have revealed that the inclusion of 10% WFS in the diet has no significant effect on
the performance of broilers. In addition, Gonzalez-Esquerra and Leeson[22] have concluded that single-comb White
Leghorn roosters tolerate up to 10% WFS in the diet in comparison to that of broiler
chicks, which have a significantly lower tolerance, manifesting as diarrhea.
Similarly, Al-Nasser et al.[14] have reported
that 10% WFS in the diet increases n-3 PUFAs and has no undesirable impact on laying
performance. Recently, Radanović et al.[23]
have studied the impact of feeding mixtures with both 5% linseed oil and fish oil
n-3 PUFAs. They found that consuming n-3 PUFA-enriched eggs had a positive effect on
some blood biochemical indicators compared to that of feeding diets with 5% soybean
oil. In addition, Aguillón-Páez et al.[24]
have observed that dietary treatments with full-fat sunflower and full-fat flaxseed
or seeds have no significant effect on egg quality parameters; however, the rate of
lay and total egg production are significantly lower in birds receiving sunflower
seeds.
Trace minerals, such as Zn, Se, and Fe, are crucial for many physiological and
biochemical functions, including production and reproduction, antioxidative status,
immune status, and lipid metabolism[25].
Optimal mineral concentrations are essential for laying hens because of their roles
in egg production, eggshell formation, egg quality, immune response, and
antioxidative properties[26]. Trace minerals
are present in the form of inorganic compounds, organic compounds, and
nano-particles. Organic sources are more bioavailable and stable than inorganic
sources, allowing the addition of feed containing lower mineral amounts[27]. In a recent study, Arbabi-Motlagh et
al.[28] have shown that organic Zn and Se
supplementation in oxidized fat diets enhances hen productive performance, egg
oxidative stability, and yolk fatty acids. To date, there has been a growing
interest in using elemental nanoparticles owing to their low toxicity, high
bioavailability, and high catalytic efficiency in broilers[29], laying hens[22],
and rabbits[30].
Therefore, the present study investigated the effects of diets containing WFS and FO
with different sources of trace elements (inorganic, organic, or nano-source of Se,
Zn, and Fe) on egg production, egg quality, sensory attributes, and plasma metabolic
profiles of laying hens.
MATERIALS AND METHODS
Ethical Approval
This study was performed using general humane care of animals that refrained from
causing suffering, distress, harm, or pain, as stated by Royal Decree number M59
in 14/9/1431H and Institutional Approval No. ACUC-22-1-2.
Experimental Design and Treatments
A total of 144 forty-week-old Hisex white-shell egg laying hens with a mean body
weight of 1703 ± 28.7 g were used. The trial lasted for 10 weeks (40–50 weeks of
age). The first 14 days (adaptation period) were not included in the study.
Throughout the study, laying hens were housed in floor pens of 1.5 × 0.6 × 2.0
m, with six hens/pen and six replicates/treatment. Each pen had a feeder and
drinker and food and water were provided ad libitum. All pens
were furnished with wood shavings and hens were kept in an environmentally
controlled room at 22−24 °C with a 16 h light /8 h dark daily cycle. The feeding
trial design consisted of six experimental treatments, each containing eight
replicates (three hens per replicate), housed in floor pens. Hens were fed six
diets as follows: 1) control diet; 2) 7.5%WFS+1.5%FO; 3) 7.5%WFS+1.5%FO+175
mg/kg vitamin E (VE); 4) 7.5%WFS+1.5%FO+175 mg/kg VE + inorganic sources of Se,
Zn, and Fe (ISeZnFe); 5) 7.5%WFS+1.5%FO+175 mg/kg VE + organic sources of Se,
Zn, and Fe (OSeZnFe); and 6) 7.5%WFS+1.5%FO+175 mg/kg VE + nano-sources of Se,
Zn, and Fe (NSeZnFe). Diets were supplemented with 0.20 mg/kg inorganic Se + 55
mg/kg inorganic Zn from Zn oxide (72% Zn) + 441 mg/kg inorganic Fe from ferrous
carbonate (40% ferrous) (WFSFOVE-Inorganic-SeZnFe), 7.5% WFS + 1.5% FO + 0.20
mg/kg organic Se (from Bio-Sel, Se yeast, Saccharomyces
cerevisiae, 0.2% with selenomethionine, min 70%) + 55 mg/kg organic
Zn from Zn chelate of glycine hydrate + 441 mg/kg organic Fe from ferrous
chelate of glycine hydrate (WFSFOVE-Organic-SeZnFe), or 7.5% flaxseed + 1.5% FO
+ 0.2 mg/kg Nano Se + 55 mg/kg Nano-Zn + 441 mg/kg Nano Fe
(WFSFOVE-Nano-SeZnFe), respectively, from a nano source containing 100%
concentration of each element. Organic Se was added as Bio-Sel 2000 Se yeast
(0.2% Se, 70% selenomethionine; IBEX International). The organic sources of Zn
and Fe were products of Wspieramy Nature (Kolejowa, Poland), as Glystatr-25% Zn
and Glystar®-Fe-17%, respectively. The nano compounds were prepared by
mechanical grinding (using a ball mill) by interaction with ascorbic acid (the
number of acid equivalents was equal to the number of salt equivalents and
therefore equal to the number of prepared nano equivalents), which performed the
bio-reduction of ions and converted them into charge less nanoparticles with
mechanical grinding that reduced the size of the nanoparticles to reach an
average size from 1–100 nm. The nanoparticles were chemically stable compounds
that were not affected by long-term storage; since the number of equivalents was
equal in the preparation, there was no increase in acid or salt during the
reaction, and therefore the concentration of the prepared nanoparticles was
100%. Since the compound was stable, it was added to the feed mixture throughout
the production cycle, but for safety, if the production cycle was 45 days, it
was added every 15 days. Whole flaxseed was purchased from a commercial supplier
and used in the experiments (Table
1). The nutritional composition and fatty acid content of the
experimental diets (Table 2) were
computed using Centraal Veevoerder Bureau feedstuff[31] analytical values and previously reported WFS chemical
assessments.
Table 1. Fatty acid profile of whole flaxseed and fish oil used in the
formulation of experimental diets for laying hens.
|
Chemical name
|
Flaxseed
|
Fish oil
|
|
Common Name
|
Systematic Name
|
%
|
%
|
| Adipic acid |
Hexanedioic acid |
- |
0.02 |
| Lauric acid |
Dodecanoic acid |
- |
0.25 |
| Tridecylic acid |
Tridecanoic acid |
- |
0.63 |
| Myristic acid |
Tetradecanoic acid |
- |
5.39 |
| Pentadecylic acid |
Pentadecanoic acid |
- |
0.98 |
| Palmitic acid |
Hexadecanoic acid |
8.90 |
11.89 |
| Margaric acid |
Heptadecanoic acid |
|
1.46 |
| Stearic acid |
Octadecanoic acid |
7.75 |
5.07 |
| Nonadecylic acid |
Nonadecanoic acid |
- |
0.44 |
| Arachidic acid |
Eicosanoic acid |
0.22 |
- |
| Heneicosylic acid |
Heneicosanoic acid |
- |
0.40 |
| Behenic acid |
Docosanoic acid |
- |
1.81 |
| Tricosylic acid |
Tricosanoic acid |
- |
0.23 |
| Lignoceric acid |
Tetracosanoic acid |
- |
0.29 |
| Cerotic acid |
Hexacosanoic acid |
- |
0.06 |
|
ΣSFA
|
16.9
|
28.9
|
| Myristoleic acid |
|
- |
0.17 |
| Palmitoleic acid |
9-Hexadecenoic acid |
- |
8.50 |
|
11-Octadecenoic acid |
- |
13. 8 |
| Oleic Acid |
9-Octadecenoic acid |
- |
6.49 |
|
cis-11-Eicosenoic
acid |
- |
3.47 |
|
13-Docosenoic acid |
- |
2.45 |
|
15-Tetracosenoic acid |
- |
1.17 |
|
6-Hexadecenoic acid |
- |
0.34 |
|
9-Eicosenoic |
- |
2.08 |
|
Petroselinic acid |
- |
0.13 |
|
ΣMUFA
|
-
|
38.6
|
|
5,11,14,17-eicosatetraenoic acid |
- |
1.05 |
| Eicosapentaenoic acid (EPA) |
cis-5,8,11,14,17-eicosapentaenoic acid |
- |
19.5 |
| Heneicosapentaenoic acid (HPA) |
6,9,12,15,18-heneicosapentaenoic
acid |
- |
0.94 |
| Docosahexaenoic acid (DHA) |
4,7,10,13,16,19-docosahexaenoic
acid |
- |
8.70 |
| α-Linolenic acid (ALA) |
9,12,15-octadecatrien-1-ol,
(Z,Z,Z) |
61.9 |
0.59 |
|
Squalene |
- |
0.13 |
|
Σn-3 PUFA
|
61.9
|
30.9
|
|
EPA+DHA
|
- |
29.2 |
| Linoleic acid |
9,12-octadecadienoic acid (Z,Z)- |
21.2 |
- |
| γ-Linolenic acid |
Gamolenic Acid |
- |
1.62 |
|
Σn-6 PUFA
|
21.2
|
1.62
|
|
Σn-6 PUFA/Σn-3
PUFA
|
0.342 |
0.052 |
|
ΣUFAs
|
83.1 |
71.1 |
|
ΣSFAs + ΣUFAs
|
100
|
100
|
ΣSFA=sum of saturated fatty acids; ΣMUFA=sum of monounsaturated fatty
acids; Σ n-3 PUFA=sum of n-3 polyunsaturated fatty acids;
EPA=eicosapentaenoic acid; DHA=ocosahexaenoic acid
Table 2. Composition and chemical analysis of experimental diets fed to
laying hens from 40–48 weeks of age.
|
Diets
|
|
Ingredients
|
Control
|
7.5%WFS+1.5%FO
|
7.5%WFS+1.5%FO+ VE
|
7.5%WFS+1.5%FO +VE+ISEZNFE1
|
7.5%WFS+1.5%FO +VE+OSEZNFE2
|
7.5%WFS+1.5%FO +VE+NSEZNFE3
|
| Yellow corn |
64.250 |
52.515 |
52.515 |
52.515 |
52.515 |
52.515 |
| Soybean meal, 48% CP |
21.50 |
20.10 |
20.10 |
20.10 |
20.10 |
20.10 |
| Corn gluten meal |
1.50 |
1.40 |
1.40 |
1.40 |
1.40 |
1.40 |
| Whole Flaxseed |
0.0 |
7.5 |
7.5 |
7.5 |
7.5 |
7.5 |
| What bran |
0.0 |
0.65 |
0.65 |
0.65 |
0.65 |
0.65 |
| Calcium diphosphate |
1.55 |
1.51 |
1.51 |
1.51 |
1.51 |
1.51 |
| Calcium carbonate |
9.00 |
9.00 |
9.00 |
9.00 |
9.00 |
9.00 |
| Sodium chloride |
0.30 |
0.30 |
0.30 |
0.30 |
0.30 |
0.30 |
| Vitamin-mineral
premix4 |
0.50 |
0.50 |
0.50 |
0.50 |
0.50 |
0.50 |
| DL-methionine |
0.10 |
0.10 |
0.10 |
0.10 |
0.10 |
0.10 |
| L-Lysine |
0.200 |
0.175 |
0.175 |
0.175 |
0.175 |
0.175 |
| Sodium bicarbonate |
0.10 |
0.10 |
0.10 |
0.10 |
0.10 |
0.10 |
| Choline chloride 50% |
0.05 |
0.05 |
0.05 |
0.05 |
0.05 |
0.05 |
| Sardine oil |
0.0 |
1.5 |
1.5 |
1.5 |
1.5 |
1.5 |
| Building sand4 |
0.95 |
4.60 |
4.60 |
4.60 |
4.60 |
4.60 |
| Vitamin E, mg/kg5 |
0.0 |
0.0 |
350 |
350 |
350 |
350 |
| Se, mg/kg |
0.0 |
0.0 |
0.0 |
4.4 |
100 |
0.20 |
| Zn, mg/kg |
0.0 |
0.0 |
0.0 |
76.4 |
220 |
55 |
| Fe, mg/kg |
0.0 |
0.0 |
0.0 |
1102.5 |
2450 |
441 |
|
Determined analysis
|
| Dry matter % |
90.87 |
90.63 |
90.57 |
91.43 |
91.13 |
90.71 |
| Crude protein % |
16.54 |
16.63 |
16.34 |
16.44 |
16.44 |
16.58 |
| Crude fat % |
2.71 |
6.21 |
6.48 |
6.51 |
6.55 |
6.63 |
| Crude fiber % |
3.27 |
3.63 |
4.13 |
4.13 |
4.13 |
4.13 |
| Ash |
16.31 |
18.93 |
15.67 |
15.81 |
15.76 |
15.68 |
|
Calculated analysis
|
| ME, kcal/kg |
2700 |
2701 |
2701 |
2701 |
2701 |
2701 |
| Ca, % |
4.04 |
4.04 |
4.04 |
4.04 |
4.04 |
4.04 |
| P available, % |
0.346 |
0.34 |
0.34 |
0.34 |
0.34 |
0.340 |
| Se, ppm |
0.367 |
0.378 |
0.378 |
0.578 |
0.578 |
0.578 |
| Zn, ppm |
93.4 |
94.4 |
94.4 |
149.4 |
149.4 |
149.4 |
| Fe, ppm |
33.3 |
40.4 |
40.4 |
481.4 |
481.4 |
481.4 |
| Methionine % |
0.507 |
0.507 |
0.507 |
0.507 |
0.507 |
0.507 |
| Lysine,% |
0.901 |
0.893 |
0.893 |
0.893 |
0.893 |
0.893 |
| C18-1, % |
0.595 |
1.166 |
1.166 |
1.166 |
1.166 |
1.166 |
| C18-2 PUFA % |
1.37 |
1.53 |
1.53 |
1.53 |
1.53 |
1.53 |
| C18-3 PUFA, % |
0.046 |
1.382 |
1.382 |
1.382 |
1.382 |
1.382 |
| C18:2/C18:3 ratio |
29.7 |
1.11 |
1.11 |
1.11 |
1.11 |
1.11 |
| Vitamin E |
25.5 |
25.5 |
200.5 |
200.5 |
200.5 |
200.5 |
1The 7.5%WFS+1.5%FO+VE+ISEZNFE-diets contain 7.5%
flaxseed + 1.5% fish oil + 175 mg/kg Vit E + 0.2 mg/kg inorganic Se as
sodium selenite (4.5% Se) + 55 mg/kg inorganic Zn from Zn oxide (72% Zn)
+ 441 mg/kg inorganic Fe from Fe carbonate (40% Fe). 2The
7.5%WFS+1.5%FO+VE+OSEZNFE-diets contain 7.5% flax seed + 1.5% fish oil +
175 mg/kg Vit E+ 0.2 mg/kg organic Se from Bio-Sel, Se yeast
(Saccharomyces cerevisiae, 0.2% with
selenomethionine, min 70%) + 55 mg/kg organic Zn from Zn chelate of
glycine hydrate + 441 mg/kg organic Fe from Fe chelate of glycine.
Organic Se was added as Bio-Sel 2000 Se yeast (0.2% Se, 70%
selenomethionine; IBEX International). The organic sources of Zn and Fe
are products of Wspieramy Nature (Kolejowa, Poland), as Glystatr-25% Zn
and Glystar®-Fe-17%, respectively. 3The
7.5%WFS+1.5%FO+VE+OSEZNFE-diets contain 7.5% flax seed + 1.5% fish oil
+ 175 mg/kg Vit E + 0.2 mg/kg nano Se + 55 mg/kg nano-Zn + 441 mg/kg
nano Fe, respectively, from nano sources containing 100% concentration
of each element. 4Building sand was added to improve crushing
of the entire flaxseed in the gizzard. 5Provided per kg diet:
Vit. A 1200000 IU/kg, Vit D3 3000.000 IU/kg, VE 25.5 mg/kg, VC 30 mg/kg,
Vit K3 (MNB) 2.05 mg/kg, Vit. B1 2.00 mg/kg, Vit. B2 6.00 mg/kg, Vit. B6
3.00 mg/ kg, Vit. B12 0.03 mg/kg, Niacin 30.00 mg/kg, folic acid 1.00
mg/kg, pantothenic acid 10.00 mg/kg, biotin 0.10 mg/kg, cobalt 0.40
mg/kg, copper 10 mg/kg, iodine 1.50 mg/kg, iron 9 mg/kg, manganese 80
mg/kg, Se 0.30 mg/kg, Zn 80 mg/kg, methionine 0.15%. 2Se, Zn,
and Fe have been added to replace the same amount of sand.
WFS = Whole flaxseed; FO = Fish oil; VE = Vitamin E (175 mg/kg); ISeZnFe
= Inorganic source of Se (0.20 mg/kg), Zn (55 mg/kg), and Fe (441
mg/kg); OSeZnFe = Organic source of Se (0.20 mg/kg), Zn (55 mg/kg), and
Fe (441 mg/kg); NSeZnFe = Nano-source of Se (0.20 mg/kg), Zn (55 mg/kg),
and Fe (441 mg/kg).
Egg weight (g), egg mass (g/hen/d), laying rate (%), feed consumption (g/hen/d),
and feed conversion ratio (FCR) (g/g) were assessed on a replicate basis and
calculated over the course of the study. At 50 weeks of age, 28 eggs were
randomly collected from each treatment on two production days (4
eggs/replicate). The quality of fresh eggs was assessed using half of the eggs,
while the other half were kept for 21 days at a temperature below 25 °C. The
eggs were then subjected to an estimation of egg quality, which included both
internal and external measurements[32,33]. Both the interior and
external qualities of eggs were evaluated[34]. Egg sensory attributes were measured (n = 7
eggs/treatment) using twenty untrained panelists to evaluate sensory
characteristics. Ten eggs from each treatment were hard-boiled for 10 min, or
until the water reached the boiling point, by covering the eggs in a saucepan of
water in a single layer. The eggs were submerged in water at 18 °C for 5 min
after the pot was removed from the hotplate. The hot water was removed and the
eggs were peeled, divided lengthwise into four equal sections, and placed on
plastic trays. The panelists rated the eggs according to their appearance, yolk
color, albumen color, flavor, and general acceptability using a continuous
unstructured line intensity scale (nine-point hedonic scale) ranging from 9
(strongly like) to 1 (strongly dislike).
Egg Nutritional Composition
A total of seven egg samples were randomly collected per treatment (one egg per
replicate) and used to determine egg chemical composition. Using the Association
of Official Analytical Chemistry techniques 934.01, 942.05, and 954.01,
respectively, the proximate analyses of eggs, including crude protein (CP),
ether extract (EE), and crude ash, were conducted[35]. The egg gross energy (GE) was determined[36]. Using the following formula, the
nitrogen-free extract (NFE) was determined as: NFE = [100 − (moisture + CP + EE
+ CF + crude ash)].
Blood Biochemical Indicators
Blood samples (n = 7) were randomly collected from each
treatment group to represent all treatment replicates for the hens at 40 weeks
of age. Blood serum was separated by centrifugation at 1500 g ×
for 10 min. The serum total protein, albumin, uric acid, and creatinine were
determined[6] using commercial kits
manufactured by Diamond Diagnostics Inc. (Holliston, MA, USA). Serum globulin
concentration = serum total protein - serum albumin. The albumin-to-globulin and
uric acid-to-creatinine ratios were calculated.
Statistical Analysis
Data were analyzed using a one-way analysis of variance statistical analysis
software program[37]. Each replicate was
considered the experimental unit. Tukey’s post-hoc test was
used to compare significant differences among the means of the treatments. All
percentage data were transformed into log10 values and normalized prior to
analysis.
RESULTS
The data presented in Table 3 show the
effects of 7.5% WFS, 1.5% FO, and different sources of Se, Zn, and Fe on laying
performance measurements in laying hens. Laying hens fed dietary treatments had no
detrimental effect on laying rate, egg weight, egg mass, feed intake (FI), FCR, body
weight change, or survival rate compared to those of hens fed the control diet.
Table 3. Effect of whole flaxseed, fish oil, and different sources of Se, Zn and
Fe fed to laying hens from 40–48 weeks of age on laying performance.
|
Dietary treatments
|
Laying rate, %
|
Egg weight, g
|
Egg mass, g/h/d
|
Feed intake, g/h/d
|
FCR, g/g
|
Body weight
change, g
|
Survival rate, %
|
| Control |
90.3 |
63.0 |
56.9 |
124.4 |
2.19 |
52.7 |
100 |
| 7.5%WFS+1.5%FO |
82.9 |
65.2 |
54.1 |
117.6 |
2.17 |
30.3 |
100 |
| 7.5%WFS+1.5%FO+VE |
91.2 |
63.8 |
58.3 |
116.1 |
1.99 |
14.0 |
100 |
| 7.5%WFS+1.5%FO+VE+ISeZnFe |
85.6 |
61.9 |
52.9 |
116.9 |
2.22 |
26.3 |
100 |
| 7.5%WFS+1.5%FO+VE+OSeZnFe |
82.1 |
63.1 |
51.9 |
122.2 |
2.38 |
76.7 |
100 |
| 7.5%WFS +1.5%FO+VE+NSeZnFe |
92.7 |
63.2 |
58.7 |
121.0 |
2.08 |
64.0 |
100 |
| RMSE |
7.61 |
1.84 |
5.04 |
8.43 |
0.254 |
78.1 |
0.0 |
| P value |
0.089 |
0.087 |
0.109 |
0.465 |
0.164 |
0.716 |
0.0 |
a–b Means within a column within each factor not sharing similar
superscripts are significantly different, P < 0.05; FCR
= feed conversion ratio; RMSE = Root mean square error; WFS = Whole
flaxseed; FO = Fish oil; VE = Vitamin E (175 mg/kg); ISeZnFe = Inorganic
source of Se (0.20 mg/kg), Zn (55 mg/kg), and Fe (441 mg/kg); OSeZnFe =
Organic source of Se (0.20 mg/kg), Zn (55 mg/kg), and Fe (441 mg/kg);
NSeZnFe = Nano-source of Se (0.20 mg/kg), Zn (55 mg/kg), and Fe (441
mg/kg).
Dietary treatments had no significant effect on egg grade, egg shape index, and shell
thickness, whereas in fresh eggs, shell percentage and shell weight per unit surface
area (SWUSA) were significantly affected by dietary treatments (Table 4). The highest shell percentage and SWUSA values
were observed in hens fed the 7.5%WFS+1.5%FO+175 mg VE/kg treatment. In stored eggs,
dietary WFS, FO, and different sources of Se, Zn, and Fe significantly affected
shell percentage, shell thickness, and SWUSA. The highest values of shell thickness
and SWUSA were observed in the 7.5%WFS+1.5%FO+VE+OSeZnFe treatment (Table 4).
Table 4. Effect of whole flaxseed, fish oil, and different sources of Se, Zn and
Fe fed to laying hens from 40–48 weeks of age on egg shape index and
eggshell quality of fresh and stored eggs.
|
Dietary treatments
|
Egg grade
|
Shape index, %
|
Shell weight, %
|
Shell thickness, µm
|
SWUSA,mg/cm2
|
|
Fresh eggs
|
| Control |
1.67 |
74.7 |
8.74a |
355 |
112.8ab |
| 7.5%WFS+1.5%FO |
1.47 |
73.6 |
8.57 a |
353 |
111.2ab |
| 7.5%WFS+1.5%FO+VE |
1.93 |
75.4 |
8.96 a |
361 |
115.0a |
| 7.5%WFS+1.5%FO+VE+ISeZnFe |
1.87 |
74.2 |
8.56 a |
346 |
110.4ab |
| 7.5%WFS+1.5%FO+VE+OSeZnFe |
2.27 |
74.5 |
8.98 a |
357 |
114.9ab |
| 7.5%WFS+1.5%FO+VE+NSeZnFe |
1.87 |
75.0 |
8.33b |
350 |
106.5b |
| RMSE |
1.076 |
3.89 |
0.622 |
22.3 |
7.97 |
| P value |
0.437 |
0.518 |
0.027 |
0.327 |
0.028 |
|
Stored eggs
|
| Control |
2.80 |
74.9 |
10.42a |
350 ab |
127.7a |
| 7.5%WFS+1.5%FO |
3.30 |
75.7 |
9.44 ab |
360 ab |
118.3 ab |
| 7.5%WFS+1.5%FO+VE |
3.40 |
80.2 |
9.96 ab |
359 ab |
124.1ab |
| 7.5%WFS+1.5%FO+VE+ISeZnFe |
3.30 |
76.3 |
9.72 ab |
347 ab |
119.6 ab |
| 7.5%WFS+1.5%FO+VE+OSeZnFe |
3.20 |
76.9 |
10.18 ab |
371a |
126.2a |
| 7.5%WFS+1.5%FO+VE+NSeZnFe |
3.10 |
76.5 |
9.18b |
341b |
112.8b |
| RMSE |
0.654 |
4.99 |
0.861 |
22.1 |
9.84 |
| P value |
0.390 |
0.257 |
0.022 |
0.050 |
0.013 |
a,b Means within a column within each factor not sharing similar
superscripts are significantly different, P < 0.05;
SWUSA = shell weight per unit of surface area; RMSE = Root mean square
error; WFS =Whole flaxseed; FO = Fish oil; VE = Vitamin E (175 mg/kg);
ISeZnFe = Inorganic source of Se (0.20 mg/kg), Zn (55 mg/kg), and Fe (441
mg/kg); OSeZnFe = Organic source of Se (0.20 mg/kg), Zn (55 mg/kg), and Fe
(441 mg/kg); NSeZnFe = Nano-source of Se (0.20 mg/kg), Zn (55 mg/kg), and Fe
(441 mg/kg).
The effects of the dietary treatments on the yolk and albumen quality characteristics
of fresh and stored eggs are listed in Tables
5 and 6, respectively. The
treatments had no significant influence on yolk percentage, albumin height, Haugh
unit (HU), or egg weight change percentage in fresh or stored eggs. However, dietary
WFS, FO, and different sources of Se, Zn, and Fe had a significant effect on yolk
color and yolk index in fresh eggs, as well as the yolk-to-albumin ratio and albumin
percentage in stored eggs. Incorporation of 7.5%WFS+1.5%FO+VE+ISeZnFe into laying
hen diets produced the greatest yolk color in fresh eggs. The
7.5%WFS+1.5%FO+VE+OSeZnFe diet produced the highest yolk index in fresh eggs.
Dietary treatment had no significant effect on egg flavor, general acceptance,
albumin color, or yolk appearance in fresh eggs (Table 7).
Table 5. Effect of whole flaxseed, fish oil, and different sources of Se, Zn and
Fe fed to laying hens from 40–48 weeks of age on egg yolk quality of fresh
and stored eggs.
|
Dietary treatments
|
Yolk weight, %
|
Yolk color
|
Yolk index, %
|
Yolk to albumin ratio
|
|
Fresh eggs
|
| Control |
26.9 |
8.07ab |
43.7 ab |
0.418 |
| 7.5%WFS+1.5%FO |
25.8 |
7.93ab |
43.7 ab |
0.393 |
| 7.5%WFS+1.5%FO+VE |
26.6 |
8.20ab |
42.6 ab |
0.413 |
| 7.5%WFS+1.5%FO+VE+ISeZnFe |
26.8 |
9.07a |
40.3b |
0.415 |
| 7.5%WFS+1.5%FO+VE+OSeZnFe |
26.5 |
7.46ab |
47.4a |
0.411 |
| 7.5%WFS+1.5%FO+VE+NSeZnFe |
26.2 |
7.13b |
46.4ab |
0.400 |
| RMSE |
1.71 |
1.52 |
0.146 |
0.0.35 |
| P value |
0.439 |
0.018 |
0.043 |
0.536 |
|
Stored eggs
|
| Control |
35.3 |
9.70 |
66.3 |
0.650a |
| 7.5%WFS+1.5%FO |
30.6 |
9.80 |
64.1 |
0.511b |
| 7.5%WFS+1.5%FO+VE |
32.7 |
10.11 |
63.7 |
0.571b |
| 7.5%WFS+1.5%FO+VE+ISeZnFe |
32.2 |
10.20 |
62..3 |
0.55 b |
| 7.5%WFS+1.5%FO+VE+OSeZnFe |
30.9 |
9.70 |
65.2 |
0.525b |
| 7.5%WFS+1.5%FO+VE+NSeZnFe |
35.8 |
9.70 |
62.1 |
0.651a |
| RMSE |
3.80 |
0.705 |
0.037 |
0.069 |
| P value |
0.112 |
0.415 |
0.529 |
0.023 |
a,b Means within a column within each factor not sharing similar
superscripts are significantly different, P < 0.05; RMSE
= Root mean square error; WFS =Whole flaxseed; FO = Fish oil; VE = Vitamin E
(175 mg/kg); ISeZnFe = Inorganic source of Se (0.20 mg/kg), Zn (55 mg/kg),
and Fe (441 mg/kg); OSeZnFe = Organic source of Se (0.20 mg/kg), Zn (55
mg/kg), and Fe (441 mg/kg); NSeZnFe = Nano-source of Se (0.20 mg/kg), Zn (55
mg/kg), and Fe (441 mg/kg).
Table 6.
Effect of whole flaxseed, fish oil, and
different sources of Se, Zn and Fe fed to laying
hens from 40–48 weeks of age on the albumin
quality traits of fresh and stored
eggs.
| Dietary treatments | Albumin, % | Albumin height, mm | Haugh unit score | Egg weight change,
% |
| Fresh eggs |
| Control | 64.3 | 5.72 | 69.8 | - |
| 7.5%WFS+1.5%FO | 65.6 | 6.44 | 75.4 | - |
| 7.5%WFS+1.5%FO+VE | 64.4 | 5.19 | 64.5 | - |
| 7.5%WFS+1.5%FO+VE+ISeZnFe | 64.6 | 5.43 | 65.8 | - |
| 7.5%WFS+1.5%FO+VE+OSeZnFe | 64.5 | 4.59 | 56.9 | - |
| 7.5%WFS+1.5%FO+VE+NSeZnFe | 65.5 | 5.45 | 65.3 | - |
| RMSE | 1.82 | 2.09 | 22.5 | - |
| P value | 0.216 | 0.273 | 0.378 | - |
| Stored eggs |
| Control | 54.3b | 3.34 | 49.8 | -9.90 |
| 7.5%WFS+1.5%FO | 59.9a | 2.63 | 37.9 | -8.94 |
| 7.5%WFS+1.5%FO+VE | 57.3ab | 2.41 | 35.7 | -8.69 |
| 7.5%WFS+1.5%FO+VE+ISeZnFe | 58.1ab | 2.08 | 32.8 | -9.39 |
| 7.5%WFS+1.5%FO+VE+OSeZnFe | 58.9ab | 2.42 | 35.1 | -9.66 |
| 7.5%WFS+1.5%FO+VE+NSeZnFe | 55.0ab | 2.63 | 41.5 | -10.50 |
| RMSE | 3.92 | 0.984 | 15.7 | 2.97 |
| P value | 0.015 | 0.122 | 0.196 | 0.782 |
a–c Means within a column
within each factor not sharing similar superscripts are significantly different,
P < 0.05; RMSE = Root mean square error; WFS = Whole
flaxseed; FO = Fish oil; VE = Vitamin E (175 mg/kg); ISeZnFe = Inorganic source of
Se (0.20 mg/kg), Zn (55 mg/kg), and Fe (441 mg/kg); OSeZnFe = Organic source of Se
(0.20 mg/kg), Zn (55 mg/kg), and Fe (441 mg/kg); NSeZnFe = Nano-source of Se (0.20
mg/kg), Zn (55 mg/kg), and Fe (441 mg/kg).
Table 7. Effect of whole flaxseed, fish oil, and different sources of Se, Zn
and Fe fed to laying hens from 40–48 weeks of age on the egg sensory
attributes of fresh eggs.
|
Dietary treatments
|
Yolk appearance
|
Albumen color
|
Egg flavor
|
Overall acceptance
|
| Control |
7.62 |
7.19 |
7.13 |
7.63 |
| 7.5%WFS+1.5%FO |
7.37 |
7.75 |
7.63 |
7.94 |
| 7.5%WFS+1.5%FO+VE |
7.56 |
7.69 |
7.00 |
7.19 |
| 7.5%WFS+1.5%FO+VE+ISeZnFe |
7.81 |
7.63 |
7.25 |
7.56 |
| 7.5%WFS+1.5%FO+VE+OSeZnFe |
7.69 |
7.63 |
7.50 |
7.57 |
| 7.5%WFS+1.5%FO+VE+NSeZnFe |
7.31 |
7.19 |
6.94 |
7.20 |
| RMSE |
1.45 |
1.48 |
1.48 |
1.39 |
| P value |
0.921 |
0.795 |
0.738 |
0.646 |
a–c Means within a column within each factor not sharing similar
superscripts are significantly different, P < 0.05; RMSE
= Root mean square error; WFS = Whole flaxseed; FO = Fish oil; VE = Vitamin
E (175 mg/kg); ISeZnFe = Inorganic source of Se (0.20 mg/kg), Zn (55 mg/kg),
and Fe (441 mg/kg); OSeZnFe = Organic source of Se (0.20 mg/kg), Zn (55
mg/kg), and Fe (441 mg/kg); NSeZnFe = Nano-source of Se (0.20 mg/kg), Zn (55
mg/kg), and Fe (441 mg/kg).
The results presented in Table 8 show
that dietary WFS, FO, and different sources of Se, Zn, and Fe significantly affected
egg chemical composition. The highest values of chemical composition were observed
in the 7.5%WFS+1.5%FO+VE+NSeZnFe treatment.
Table 8. Effect of whole flaxseed, fish oil, and different sources of Se, Zn and
Fe fed to laying hens from 40–48 weeks of age on chemical composition of
whole edible egg parts.
|
Dietary treatments
|
Dry matter, %
|
Crude protein, %
|
Ether extract, %
|
Ash, %
|
NFE, %
|
GE, cal/g
|
| Control |
50.6b |
17.0ab |
31.3b |
0.971 |
0.800 |
350d |
| 7.5%WFS+1.5%FO |
50.6ab |
16.8ab |
31.6ab |
0.895 |
0.800 |
369c |
| 7.5%WFS+1.5%FO+VE |
50.4ab |
16.5b |
31.7ab |
0.912 |
0.803 |
388b |
| 7.5%WFS+1.5%FO+VE+ISeZnFe |
50.7ab |
16.8ab |
31.7ab |
0.917 |
0.813 |
392ab |
| 7.5%WFS+1.5%FO+VE+OSeZnFe |
50.5ab |
16.6b |
31.7ab |
0.926 |
0.788 |
397ab |
| 7.5%WFS+1.5%FO+VE+NSeZnFe |
51.5a |
17.3a |
32.1a |
0.927 |
0.802 |
400a |
| RMSE |
0.481 |
0.309 |
0.413 |
0.059 |
0.021 |
5.97 |
| P value |
0.004 |
0.002 |
0.105 |
0.375 |
0.518 |
<.0001 |
a,b,c Means within a column within each factor not sharing similar
superscripts are significantly different, P < 0.05; NFE
= nitrogen free extract; GE = gross energy; RMSE = Root mean square error;
WFS = Whole flaxseed; FO = Fish oil; VE = Vitamin E (175 mg/kg); ISeZnFe =
Inorganic source of Se (0.20 mg/kg), Zn (55 mg/kg), and Fe (441 mg/kg);
OSeZnFe = Organic source of Se (0.20 mg/kg), Zn (55 mg/kg), and Fe (441
mg/kg); NSeZnFe = Nano-source of Se (0.20 mg/kg), Zn (55 mg/kg), and Fe (441
mg/kg).
As shown in Table 9, different levels of
FSC have no effect on serum protein metabolites, uric acid, creatinine, or their
ratios. The highest plasma globulin concentrations were observed in hens fed the
7.5%WFS+1.5%FO+VE and 7.5%WFS+1.5%FO+VE+NSeZnFe treatments. However, the lowest
plasma uric acid concentrations were observed in hens fed the
7.5%WFS+1.5%FO+VE+OSeZnFe and 7.5%+1.5%FO+VE+NSeZnFe treatments.
Table 9. Effect of whole flaxseed, fish oil, and different sources of Se, Zn and
Fe fed to laying hens from 40–48 weeks of age on the levels of serum protein
fractions.
|
Dietary treatments
|
Total protein, g/dL
|
Albumin, g/dL
|
Globulin, g/dL
|
Albumin/globulin
|
Creatinine, mg/dL
|
Uric acid, mg/dL
|
Uric acid/creatinine
ratio
|
| Control |
5.23a |
3.73a |
1.50b |
2.48a |
1.054a |
3.01a |
2.86abc |
| 7.5%WFS+1.5%FO |
5.22a |
3.72a |
1.50b |
2.48a |
1.055a |
3.00bc |
2.84d |
| 7.5%WFS+1.5%FO+VE |
5.21b |
3.70b |
1.52a |
2.45b |
1.045b |
2.99bc |
2.87a |
| 7.5%WFS+1.5%FO+VE+ISeZnFe |
5.20c |
3.69c |
1.49b |
2.47a |
1.044b |
3.00b |
2.87a |
| 7.5%WFS+1.5%FO+VE+OSeZnFe |
5.19c |
3.71b |
1.49b |
2.50a |
1.049b |
2.98c |
2.85cd |
| 7.5%WFS+1.5%FO+VE+NSeZnFe |
5.21b |
3.71b |
1.51a |
2.44b |
1.047b |
2.98c |
2.85bc |
| RMSE |
0.008 |
0.004 |
0.007 |
0.013 |
0.004 |
0.006 |
0.011 |
| P value |
<.0001 |
<.0001 |
<.0001 |
<.0001 |
<.0001 |
<.0001 |
<.0001 |
a–d Means within a column within each factor not sharing similar
superscripts are significantly different, P < 0.05; RMSE
= Root mean square error; WFS = Whole flaxseed; FO = Fish oil; VE = Vitamin
E (175 mg/kg); ISeZnFe = Inorganic source of Se (0.20 mg/kg), Zn (55 mg/kg),
and Fe (441 mg/kg); OSeZnFe = Organic source of Se (0.20 mg/kg), Zn (55
mg/kg), and Fe (441 mg/kg); NSeZnFe = Nano-source of Se (0.20 mg/kg), Zn (55
mg/kg), and Fe (441 mg/kg).
DISCUSSION
The findings presented in Table 3 show
that laying hens fed 7.5%WFS, 1.5%FO, and different sources of Se, Zn, and Fe in
their diets have no detrimental effects on laying performance compared to those of
hens fed the control diet. These results are consistent with those of previous
studies[1,38,39], which show that laying
hens withstand up to 10% WFS in the diet without adversely affecting laying
performance indicators. Bean and Leeson[38]
have shown that 10% dietary WFS has no significant effect on egg weight or
production in brown and white laying hens. Similarly, Scheideler and Froning[40] have confirmed that laying hens tolerate up
to 15% dietary WFS without any negative effects on egg-laying performance. Mattioli
et al.[39] have stated that 10% extruded
flaxseed in the diet does not affect egg-laying performance, including egg-laying
rate, egg weight, egg yield, FCR, or final body weight of laying hens. Hosseini et
al.[41] have assumed that feeding with
flaxseed (270, 180, 90, and 0 g/kg) has no significant effect on egg mass, FI, or
FCR. Therefore, it was reasonable to predict that dietary WFS had no negative
influence on yolk formation or egg production rates. The percentage of WFS used in
the diet may be increased up to 15% using additional treatments, such as adding
enzyme mixtures or heat. Moghadam et al.[42]
have observed that 15% heated flaxseed in the diet enhances hen day egg production
and egg mass, whereas FI and egg weight are not affected. Jia et al.[43] have shown significant improvements in egg
production in hens fed 15% flaxseeds and multi-carbohydrate enzymes. Aguillón-Páez
et al.[24] have stated that dietary inclusion
of full-fat flaxseed seeds increases egg quality compared to that of hens fed
full-fat sunflower seeds.
The different sources of Se, Zn, and Fe had no detrimental effects on laying
performance. These data are consistent with those reported in several previous
studies[44,45,46]. Kim et al.[45] have illustrated that the addition of an
inorganic and organic mineral premix has no negative influence on egg weight, hen
day egg production, FI, or FCR compared to those of hens fed the control diet. In
addition, Kannan et al.[46] have shown that
dietary inorganic and organic Fe at 100, 200, and 300 mg/kg has no significant
effect on egg production rate, FI, or egg weight in laying hens. In a study,
Buckiūnienė et al.[44] have concluded that
different dietary Fe sources have no significant effect of laying performance,
including rate of egg production, FI and FCR in laying hens. Estradiol-17β
stimulates yolk lipoprotein precursor synthesis, such as vitellogenin and
very-low-density lipoprotein (VLDL) in the liver, which are then absorbed by the
developing yolk via an endocytosis mediated receptor[47]. Plasma Zn levels indicate vitellogenin in the bloodstream[48]. The present study found that the
incorporation of 7.5%WFS, 1.5%FO, and different sources of Se, Zn, and Fe did not
alter plasma levels of VLDL and Zn (data not shown). Therefore, dietary
7.5%WFS+1.5%FO+VE+OSeZnFe did not negatively affect folliculogenesis or the growth
of ovarian follicles, which, in turn, was associated with the rate of egg production
in the current study.
Dietary treatment had no effect on external egg quality features, including egg
grade, egg shape index, and shell thickness (Table 4). These results correspond with those of Mattioli et al.[39], who have shown that the inclusion of 10%
extruded flaxseed in the diet has no significant effect on shell quality traits.
Bean and Leeson[38] have observed that 10%
WFS in the diet has no significant effect on shell weight, albumin height, or shell
thickness in laying hens. Likewise, Hosseini et al.[41] have shown that dietary extruded flaxseed at–0–2700 g/kg in 90 g/kg
increments has no effect on shell thickness or strength. Moghadam et al.[42] have observed that 15% heated flaxseed in
the diet has no significant effect on shell weight or thickness. Similarly, Panaite
et al.[49] have shown that 5% WFS
supplemented with different levels of dried tomato waste (2.5%, 5.0%, and 10.0%) in
the diet does not affect shell weight, shell thickness, or shell-breaking strength
in laying hens. These data suggest that WFS had no negative effects on eggshell
formation and did not interfere with the supply of calcium required for shell
formation. This assumption has been confirmed by the results of Caston et al.[50], who have postulated that 20% WFS in the
diet has no influence on eggshell deformation or absolute shell weight. In addition,
Ebeid[34] and Bozkurt et al.[51] have reported similar outcomes when hens
are fed flaxseed oil or FO as a n-3 PUFAs source. In the literature, different
sources of Fe[44] and Zn[27] have no significant effects on external egg
quality parameters or sensory attributes. However, Ogbuewu and Mbajiorgu[52] have indicated that dietary Zn
supplementation enhances eggshell thickness in laying hens. Arbabi-Motlagh et
al.[28] have reported that dietary
organic Zn (20 mg/kg) plus organic Se (0.2 mg/kg) in oxidized fat diets is
beneficial for improving shell quality characteristics (shell thickness and shell
breaking strength) in laying hens.
The results in Tables 5 and 6 show that dietary treatments have no
significant effect on yolk percentage, albumin height, HU, or egg weight change
percentage in both stored and fresh eggs. These results are in agreement with
previous findings[2,50] reporting that dietary WFS has no negative effect on yolk
weight or relative yolk weight. Furthermore, adding 10% WFS to the diets of laying
hens does not significantly influence the albumin percentage, albumen height, or HU
score, according to other studies[38,53]. Recently, Hosseini et al.[41] have postulated that graded amounts of
dietary extruded flaxseed have no significant effect on the HU score. Similarly,
Mattioli et al.[39] have shown that 10%
extruded flaxseed in the diet does not affect the HU score, whereas the percentage
of albumin increased. Additionally, 15% heated flaxseed in the diet has no
significant effect on HU, albumin weight and thickness, or yolk weight, height, and
width[42].
As shown in Table 5, dietary WFS, FO, and
different sources of Se, Zn, and Fe have a significant effect on yolk color and yolk
index in fresh eggs, as well as the yolk-to-albumin ratio and albumin percentage in
stored eggs. Incorporation of 7.5%+1.5%FO+VE+ISeZnFe into laying hen diets produced
the greatest yolk color in fresh eggs. The 7.5%+1.5%FO+VE+OSeZnFe diet produced the
highest yolk index in fresh eggs. Similarly, Westbrook and Cherian[54] have revealed that the addition of a
0.05%–0.1% enzyme mixture to 10% flaxseed diet enhances yolk weight. The yolk color
results are in agreement with those of Moghadam et al.[42], who have observed that 15% heated flaxseed in the diet
improves yolk color score, and this improvement may be related to enhanced
digestibility and consequently more utilization of antioxidants and pigmentation in
flaxseeds. Furthermore, dietary flaxseed oil and/or FO supplementation significantly
positively affects egg yolk color[34].
Cachaldora et al.[55] have reported similar
findings, demonstrating that n-3 PUFA oil-enriched diets improve yolk color compared
to that of the control diet. Dietary carotenoids are fat-soluble; therefore,
intestinal absorption is expected to increase in tandem with lipid absorption[34]. Notably, improving yolk color has a
positive effect on customer choice of eggs produced by chickens fed flaxseed, which
is an additional advantage of raising n-3 PUFA levels[50].
Dietary WFS, FO, and different sources of Se, Zn, and Fe had no significant effect on
yolk appearance, general acceptance of albumin color, or flavor in fresh eggs. These
results agree with those of Hayat et al.[56],
who have revealed that adding 10% WFS to the diet of laying hens has no adverse
effects on egg sensory characteristics and that eggs are acceptable to untrained
panelists and consumers. In addition, Aguillón-Páez et al.[24] have reported that dietary inclusion of full-fat sunflower
or full-fat flaxseed seeds has no significant effect on egg quality parameters.
Likewise, Buckiūnienė et al.[44] have
reported that dietary rapeseed oil plus organic or inorganic Se plus VE has no
undesirable effect on egg sensory characteristics (odor, taste, color, and
acceptability), and texture traits (hardness, cohesiveness, springiness, and
chewiness). Dietary Se and Zn improve antioxidative properties and reduce lipid
peroxidation[25,26,48], which is
associated with enhanced oxidative stability of egg yolks, leading to improved
sensory attributes. Imran et al.[57] have
shown that most untrained evaluators do not discriminate between
n-3-enriched eggs and controls that come from laying hens fed a 10%
extruded flaxseed meal in their diet. Mazalli et al.[58] have reported no unpleasant fishy flavor in hard-boiled eggs
obtained from chickens fed 9% WFS. Professional panelists, however, detect
variations in organoleptic properties in eggs produced by laying hens fed flaxseed
meal, especially at high levels[57]. Hence,
it is critical to emphasize the importance of careful consideration when using WFS
in the diet of laying hens.
The findings presented in Table 8 show
that dietary WFS, FO, and different sources of Se, Zn, and Fe have significant
effects on the chemical composition of eggs, including dry matter, CP, EE, and GE.
However, few studies have investigated this aspect. Mattioli et al.[39] have observed that 10% extruded flaxseed in
the diet has no influence on egg chemical proximate analyses, including dry matter,
EE, ash, and CP, whereas egg cholesterol content is reduced.
As shown in Table 9, different levels of
FSC have a significant effect on serum protein metabolites, uric acid, creatinine,
and the uric acid:creatinine ratio. Similarly, Hosseini et al.[41] have hypothesized that adding extruded flaxseed to the diet
has no significant effect on total protein, albumin, globulin, or albumin:globulin
ratio in hens. In addition, Shafey et al. [59] have observed that adding 5% or 10% flaxseed meal to laying hen diets
have an insignificant effect on serum protein metabolites in laying hens. The
current study found that the highest plasma globulin concentrations were observed in
hens fed the 7.5%WFS+1.5%FO+VE and 7.5%WFS+1.5%FO+VE+NSeZnFe treatments. These
results may be attributed to the role of trace mineral nanoparticles in improving
protein and immunoglobulin synthesis[60].
In conclusion, feeding laying hens up to 7.5% WFS had no negative effects on laying
performance indices, quality characteristics, sensory attributes, chemical
composition of eggs, or blood plasma biochemical parameters. Furthermore,
7.5%WFS+1.5%FO+VE+OSeZnFe in the diet enhanced shell thickness, SWUSA, yolk color,
yolk index, and plasma globulin concentration in laying hens.
ACKNOWLEDGMENTS
This research was funded by Researchers Supporting Project No. RSPD2024R581, King
Saud University, Riyadh, Saudi Arabia.
Author Contributions
Conceptualization: Y.A.A.; methodology: Y.A.A., M.A.-H., A.A.Al-S., EL-S. O.S.H.,
and M.J.O.; software: Y.A., N.M.A., and M.J.O.; data collection: M.J.O., EL-S.
O.S.H., and A.A.Al-S.; investigation: Y.A.A., M.J.O., N.M.A., A.A.Al-S., M.J.O.,
G.M.S., EL-S., and O.S.H; resources: A.A.Al-S., EL-S., O.S.H., T.A.E., and M.J.O.;
writing—original draft preparation: Y.A.A., T.A.E., N.M.A., and M.J.O.;
writing—review and editing: Y.A.A., N.M.A., M.J.O., T.A.E., V.T., M.R., and
A.A.Al-S.; project administration and supervising: Y.A.A.; funding acquisition:
Y.A.A. All the authors have read and agreed to the published version of the
manuscript.
Conflicts of Interest
The authors declare no conflict of interest.
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