Feeding high-oleic peanuts to layer hens enhances egg yolk color and oleic

fatty acid content in shell eggs

Ondulla T. Toomer,∗,1 Amanda M. Hulse-Kemp,† Lisa L. Dean,∗ Deborah L. Boykin,‡

Ramon Malheiros,§ and Kenneth E. Anderson§
∗
U.S. Department of Agriculture, Agricultural Research Service, Market Quality & Handling Research Unit,
Raleigh, NC 27695, USA; † U.S. Department of Agriculture, Agricultural Research Service, Genomics and
Bioinformatics Research Unit, Raleigh, NC 27695, USA; ‡ U.S. Department of Agriculture, Agricultural Research
Service, Office of the Area Director, Stoneville, MS 38776, USA; and § Department of Poultry Science, North
Carolina State University, Raleigh, NC 27695, USA

ABSTRACT Previous studies have identified normal-
oleic peanuts as a suitable and economical broiler feed
ingredient. However, no studies to date have examined
the use of high-oleic (HO) peanut cultivars as a feed
ingredient for laying hens and determined the impact
of feeding HO peanuts on performance and egg nutri-
tive qualities. This project aimed to examine the use of
HO peanuts as a feed ingredient for layer hens to de-
termine the effect on performance, egg lipid chemistry,
and quality of the eggs produced. Forty-eight 40-wk-
old layer hens were fed a conventional soybean meal
+ corn control diet or a HO peanut + corn diet for
10 wk in conventional battery cages. Body and feed
weights were collected weekly. Pooled egg samples were
analyzed for quality, lipid analysis, and peanut protein
allergenicity. There were no significant differences in
hen performance or egg quality as measured by USDA
grade quality, egg albumen height, or egg Haugh unit
between the treatment groups. However, eggs produced
Key words: high-oleic peanuts, feed ingredients, layer hens, shell eggs, β -carotene
from layer hens fed the HO peanut + corn diet had re-
duced egg weights relative to the controls (P = 0.0001).
Eggs produced from layer hens fed the HO peanut diet
had greater yolk color scores (P < 0.0001), HO fatty
acid (P < 0.0001), and β -carotene (P < 0.0001) levels
in comparison to the controls. Eggs produced from hens
fed the control diet had greater palmitic and stearic
saturated fatty acids (P < 0.0001), and trans fat (P <
0.0001) content compared to eggs produced from hens
fed the HO peanut diet. All egg protein extracts from
all treatments at each time point were non-reactive with
rabbit anti-peanut agglutinin antibodies. This study
identifies HO peanuts as an abundant commodity that
could be used to support local agricultural markets of
peanuts and poultry within the southeastern United
States and be of economic advantage to producers while
providing a potential health benefit to the consumer
with improved egg nutrition.
2019 Poultry Science 98:1732–1748
http://dx.doi.org/10.3382/ps/pey531

INTRODUCTION tities of these grains and high protein oilseeds are

For decades, the poultry industries (broilers, eggs,
turkeys) have been of great economic importance and
represented a significant portion of the agricultural
products sold in the southeastern (Georgia, Arkansas,
Alabama, North Carolina, South Carolina, and Mis-
sissippi) United States (APHIS, USDA, 2015). Within
this region, the need for poultry feed components such
as corn and soybean meal far exceeds the ability to
produce these ingredients locally. Hence, large quan-
Published by Oxford University Press on behalf of Poultry Science
Association 2018. This work is written by (a) US Government em-
ployee(s) and is in the public domain in the US.
Received April 23, 2018.
Accepted October 30, 2018.
1
Corresponding author: Ondulla.Toomer@ars.usda.gov
imported from South America (Economic Research Ser-
vice, USDA, 2017) and routinely from Iowa and Illinois,
the top producing soybean and corn states (Agweek,
2009) in the U.S. Mid-West for animal feed produc-
tion. This study aims to examine the use of high-oleic
peanuts, a valuable oilseed crop abundantly grown also
within the U.S. southeast, with Georgia producing ap-
proximately 46% of U.S. peanuts annually (National
Peanut Board, 2018).
In various parts of the world, peanut meal from
normal-oleic peanuts (groundnuts) is utilized as a com-
mon protein source for feeding livestock (Cilly et al.,
1977; Olomu and Offiong, 1980, 1985; Aletor and
Olonimoyo, 1992; Venkataraman et al., 1994; Donkoh
et al., 1999; Naulia and Singh, 2002). Yet, in the United
States 80% of peanut production is used for peanut but-
ter, oil production, and snacks for human consumption

1732
 EGGS FROM LAYER HENS FED HIGH-OLEIC PEANUTS
(American Peanut Council, 2017). Although earlier
research studies demonstrated that peanut meal
prepared from normal-oleic peanuts is a suitable and
economical poultry feed ingredient (Costa et al., 2001;
Pesti et al., 2003), few studies have examined the use
of modern high-oleic peanut cultivars (80% oleic fatty
acids and 2% linoleic fatty acids) as a feed ingredient
for laying hens. In addition, early poultry feeding
studies using normal-oleic peanuts determined lysine
as the first limiting dietary amino acid, followed by
methionine and tryptophan (Douglas and Harms,
1959; Waldroup and Harms, 1963). However, relatively
recent introduction of affordable purified threonine and
other amino acids has made the inclusion of purified
amino acids in poultry rations economically reasonable.
Studies by Moreira et al. (2014, 2016) demonstrated
the human health benefits of moderate consumption of
high-oleic peanuts; however, few studies have examined
how feeding peanuts high in monounsaturated fatty
acids to egg-producing hens and poultry may alter the
fatty acid and lipid profile of poultry meat and eggs
produced. Studies by Shapira et al. (2008) demon-
strated that an extruded linseed-supplemented diet fed
to layer hens increased the omega-3 polyunsaturated
fatty acid (PUFA) content in table eggs compared to
control eggs, with fortified eggs yielding approximately
a 3.8-fold increase in omega 3 PUFA, 6.4-fold increase
in alpha-linolenic acid (18:3), and 2.4-fold increase in
docohexaenoic acid (DHA). Moreover, laying hens
fed diets with varying levels of fish meal (0% control
diet to 20%) had increased DHA in egg yolk relative
to increased dietary inclusion of fish meal compared
to the controls with the exception of 15 and 10%
fish meal, which led to the same increase of DHA
levels in egg yolk (Howe et al., 2002).
However, the inclusion of greater than 1 to 2%
of marine oils (primary source PUFA) in poultry di-
ets compromises the oxidative stability of the meat
and/or eggs (Lopez-Ferrer et al., 2001; Baeza et al.,
2013), while producing off-flavors (Hargis et al., 1991).
Other reports have demonstrated that linseed inclu-
sion in the diets of hens also produces off-flavors
in the meat and eggs produced similar to those of
hens fed diets with marine oils (Woods and Fearon,
2009).
Therefore, in this study we aimed to 1) examine the
usefulness of whole high-oleic peanuts as a cost-effective
and suitable protein and energy-rich feed ingredient for
40-wk-old layer hens, and 2) determine the effects of
feeding a high-oleic peanut diet to egg-producing layer
hens on the nutritive value, lipid, and fatty acid com-
position of the shell eggs produced. We conjecture that
the eggs produced from layer hens fed a diet with high-
oleic peanuts will have improved lipid and fatty acid
profiles with enhanced healthy monounsaturated fatty
acids compared to poultry fed a conventional layer hen
diet.
1733
MATERIALS AND METHODS
Experimental Design, Animal Husbandry
and Dietary Treatments
To test this hypothesis, we compared 2 isonitrogenous
and isocaloric dietary treatments prepared 1 wk prior
to the onset of the study and maintained in feed storage
bins in North Carolina State University (NCSU) Feed
Mill in a cool dry location. Treatment 1 was a high-oleic
peanut + corn diet (18% crude protein, 11% fat, and
1,400 metabolizable energy) prepared using aflatoxin-
free whole non-roasted high-oleic peanuts with the testa
(skin) intact crushed using a Roller Mill. Prior to in-
clusion within the diet, whole raw high-oleic peanuts
were crushed using a Roller Mill. The high-oleic peanut
+ corn diet (HO PN) was composed predominately of
yellow corn, corn gluten, whole high-oleic peanuts (non-
roasted, testa intact), and wheat bran, supplemented
with amino acids L-lysine, L-methionine, L-tryptophan,
L-threonine along with NCSU vitamin and mineral pre-
mix (Figure 1). Treatment 2 was a soybean meal +
corn diet (SBM), a control conventional mash diet (18%
crude protein, 7% fat, and 1,400 metabolizable energy),
composed predominately of yellow corn, corn gluten,
soybean meal, wheat bran, and poultry fat, with NCSU
vitamin and mineral premix (Figure 1). Experimental
diets were analyzed and determined to be free of afla-
toxin and microbiological contaminants by the North
Carolina Department of Agriculture and Consumer Ser-
vices, Food and Drug Protection Division Laboratory
(Raleigh, NC).
Forty-eight 40-wk-old White Leghorn layer hens
(NCSU University maintained Poultry Flock) were ran-
domly assigned to 24 animals per treatment. Animals
were housed in 1 room with 2 sets of conventional bat-
tery cages housing 24 hens each on each side of the
room designated as left and right (1 hen per cage;
12 cages upper row + 12 cages lower row) with 12
animals randomly assigned to 2 replicate pens within
each treatment group. To account for treatment vari-
ability due to environmental factors such as ventilation
and/or temperature in the room, 12 animals per treat-
ment were housed on separate sides of the room to as-
certain the effect of environmental differences between
each side of the room. Animals were provided feed and
water ad libitum and housed individually with 14 h
of light daily for 10 wk. Body and feed weights were
recorded weekly. Daily shell eggs were collected, and
weights were recorded from each hen. Weekly, shell eggs
were analyzed for quality (yolk color DSM, albumen
height, Haugh unit [HU]) and USDA grading by the
Layer Hen & Small Flock Management Lab, Depart-
ment Poultry Science, NCSU. The Institutional Ani-
mal Care and Use Committee at NCSU approved all
experimental animal protocols prior to the onset of this
study.
1734 TOOMER ET AL.

Figure 1. Layer hen dietary treatments and composition. (A) Treatment 1 was a high-oleic peanut + corn diet (HOPN: 18% crude protein,
11% fat, and 1,400 metabolizable energy) composed predominately of yellow corn, corn gluten, whole high-oleic peanuts with the skins intact (non-
roasted) and wheat bran, supplemented with amino acids L-lysine, L-methionine, L-tryptophan, L-threonine along with vitamin and mineral
premix. Prior to inclusion within the diet, whole raw high-oleic peanuts were crushed using a Roller Mill. (B) Treatment 2 was a control,
conventional soybean meal + corn layer hen mash diet (SBM:18% crude protein, 7% fat, and 1,400 metabolizable energy (kCal/lb.)) composed
predominately of yellow corn, corn gluten, soybean meal, wheat bran and poultry fat, with vitamin and mineral premix. (C) Composition table
of experimental diets in percent by weight.

Layer Hen Feed Analysis
Total nitrogen was determined in homogenized sam-
ples by combustion using an Elementar N cube ana-
lyzer (Elementar Americas, Mt. Laurel, PA) according
to AOAC 990.03 (2007). A Kjeldahl conversion factor
of 6.25 (mixed food) was used to calculate total protein
in the samples.
Total fat as triglycerides was determined in the sam-
ples gravimetrically after Soxhlet extraction. Samples
were extracted for 6 h using continuous extraction with
hexane (AOAC 920.39, 1990).
Egg Quality and Grading
Albumen height and HU (Haugh, 1937) were
recorded with the TSS QCD system (Technical Services
and Supplies, Dunnington, York, UK). Egg albumen
height and HU were calculated to determine egg albu-
men quality. Yolk color was determined by using TSS
QCD System yolk color scan which is calibrated to the
DSM Color Fan, which consists of a series of 15 col-
ored plastic tabs arranged as a fan corresponding to the
range of yolk colors found in yellow from light yellow to
orange-red (a color index 1 to 15 to distinguish the yolk
color density from lightest to darkest color intensity),
earlier defined by Vuilleumier (1969).
Eggs were visually inspected for exterior and inte-
rior grading in accordance with the USDA Standards
(USDA 2010). The exterior of eggs was examined to
ensure that a clean, smooth, defect-free, oval shape sur-
face was present, lacking thin spots or textured appear-
ance. Interior grading was conducted by the candling
method to examine the air cell, the albumen, and yolk
and to examine for meat and blood spots and cracks.
Eggs were sized weekly by treatment (USDA, 2010).
Egg sizing was classified by a minimum net weight per
egg: peewee (<42.6 g), small (42.6 < 56.8 g), medium
(49.7 < 56.8 g), large (56.8 < 63.9 g), extra-large
(63.9 < 70.9 g), and jumbo (>70.9 g).
Egg Samples—Protein Extraction
Shell eggs were collected, labeled, and weighed daily
from each laying hen. All eggs produced by an in-
dividual layer hen were pooled in a sterile Whirl-
pak bag (Thermo Fisher Scientific, Rockford, IL) and
homogenously mixed using a Stomacher Lab Blender.
 EGGS FROM LAYER HENS FED HIGH-OLEIC PEANUTS
Total proteins were extracted from pooled shell egg
samples (experimental weeks 1, 3, 5, 7, and 10) in
20 mM Tris-HCL pH 8.1 extraction buffer supple-
mented with 1% HALT protease inhibitor (Thermo
Fisher Scientific) using 1 mL of homogenous pooled
shell egg sample to 10 mL of extraction buffer. Sam-
ples were vortexed and centrifuged for 10 min at 4◦ C,
and the supernatants (protein extracts) were used for
analysis. Protein concentration of protein extracts was
determined using a BCA Protein Assay Kit (Thermo
Fisher Scientific). Protein extracts were aliquoted and
stored at –20◦ C.
SDS-PAGE Analysis and Immunoblotting
Protein extracts from pooled shell egg samples (75 μg
per lane) were loaded on a 10% polyacrylamide Tris-
glycine gel (Criterion TGX, Bio-Rad, Hercules, CA)
and electrophoretically separated under constant volt-
age with Tris/Glycine/SDS buffer according to the
manufacturer’s instructions (Bio-Rad). Proteins were
visualized by Coomassie Brilliant Blue staining and dig-
itally imaged. SDS-PAGE resolved proteins that were
transferred to nitrocellulose membranes electrophoreti-
cally for immunoblotting procedures using Midi Trans-
Blot Turbo Transfer Pack (Bio-Rad) The membranes
were washed 3 times in Tris-buffered saline with 0.1%
Tween (TBST) and subsequently blocked in 5% non-
fat dry milk (NFDM) in TBST for 1 h at room tem-
perature with shaking. To examine the potential peanut
allergenicity of protein extracts from shell egg samples,
membranes were incubated at 4◦ C overnight with shak-
ing in 5% NFDM in TBST containing rabbit IgG anti-
peanut agglutinin primary antibody 1:1000 dilution
(LifeSpan Biosciences, Seattle, WA). Membranes were
washed 3 times in TBST and incubated in 5% NFDM
in TBST containing secondary antibody donkey anti-
rabbit IgG-HRP (Santa Cruz Biotechnology, Dallas,
TX) dilution 1:1,000 at room temperature with shaking
for 1 h. Bio-detection was determined utilizing chro-
mogenic peroxidase substrate CN/DAB (chloronaph-
thol and diaminobenzidine, Thermo Fisher Scientific)
detection of HRP (horseradish peroxidase) activity.
Lipid and Fatty Acid Analysis
Lipid and fatty acid analysis (total fat, total oleic
fatty acid, and total linoleic fatty acid) of homoge-
nous pooled total egg samples (experimental weeks
2, 3, 4, 6, 7, 8, and 9) and feed samples from both
treatment groups were analyzed using direct methy-
lation modified methods as described by Wang et al.
(2000) in the Market Quality & Handling Research
Unit. Pooled total egg samples were thawed at room
temperature and vortexed to obtain a homogenous mix-
ture. One milliliter of internal standard (tridecanoic
acid, 1 mg/mL, Fluka Chemical Corp., Milwaukee, WI)
in hexane was added to a screw-capped glass tube. The
1735
hexane (Thermo Fisher Scientific, Waltham, NJ) was
evaporated in a stream of nitrogen. One hundred mil-
ligrams of pooled total egg sample was added to the
tube, and the weight was recorded. One milliliter of
methanol (Thermo Fisher) was added, and the con-
tents of the tube were vortexed. Three milliliters of
methanolic hydrochloric acid (Sigma Chemical Corp.,
St. Louis, MO) was subsequently added. The tube was
capped, vortexed, and placed in a water bath for 1 h at
95◦ C. After removing from the water bath and cooling
to room temperature, 8 mL of 0.88% sodium chloride
(Sigma) in water was added and the tube was vortexed.
Subsequently, 3 mL of hexane was added. The tube was
vortexed for 30 s and centrifuged for 15 min at 1,000 × g
using the IEC Model K centrifuge. The top organic
layer was transferred to a clean glass tube containing
approximately 50 mg of sodium sulfate (Sigma) to dry
the solvent. The layer was then transferred to a crimp
top autosampler vial. The samples were run on the PE-
AS XL Autosampler GC (Perkin Elmer, Shelton, CT)
with Kel Fir Fame 5 and Kel Fir Fame 6 Standards (Ma-
treya LLC, State College, PA). The column used was
an SGE BPX-70 (70% Cyanopropyl Polysilphenylene-
siloxane, SGE Analytical Science, Austin, TX, 30 m
length, 0.25 mm i.d., 0.25μ df). The temperature pro-
gram was 60◦ C for 2-min hold, 10◦ C per minute to 180◦ ,
0-min hold, and then 4◦ C per minute to 235◦ C with
0-min hold for a total of 27.75 min. The injector was
split at 40 mL/min at 220◦ C. The detector was FID
at 250◦ C. The carrier gas was helium at 1.85 mL/min.
Fatty acids were identified by retention time matches
using the Kel Fir Fame 6 Standard. The results were
calculated as total fat (g/100 g) as triglycerides using
response factors calculated from the Kel Fir Fame 5
Standard (Ngeh-Ngwainbi et al., 1997).
Homogeneous egg samples from each treatment
group were chemically analyzed for crude fat,
cholesterol, β -carotene, and 36 differing fatty acids
(butyric, caproic, caprylic, undecanoic, lauric, tride-
canoic, myristic, myristoleic, pentadecylic, pentade-
cenoic, palmitic, palmitoleic, margaric, margaroleic,
stearic, oleic, n9 t elaidic, linoleic, linolenic, n6
gamma-linolenic, arachidic, gadoleic, eicosadienoic,
homo-gamma-linolenic, eicosatrienoic, arachiconic, n3
timnodonic, heneicosanic, behenic, erucic, brassic, do-
cosahexaenoic, lignoceric, nervonic, omega 3, and
omega 6) content at weeks 1, 5, and 10 experimentally
(Tables 5, 6, 8, and 9). Analysis of 36 differing fatty
acids on all 775 egg samples was cost-prohibitive at
all experimental time points (week 1 to 10). Therefore,
only eggs produced at weeks 1, 5, and 10 were analyzed
for the full panel of 36 fatty acids by an external vendor
(ATC Scientific).
For β -carotene and comprehensive lipid and fatty
acid analysis, ATC Scientific (Little Rock, AR) ana-
lyzed pooled total egg samples for experimental weeks
1, 5, and 10 of experimental samples only due to
cost restrictions. Pooled total egg samples were ana-
lyzed for the following lipids and fatty acids: crude fat,
1736 TOOMER ET AL.
butyric acid, caproic acid, caprylic acid, capric acid,
lauric acid, tridecanoic acid, myristic acid, myristoleic
acid, pentadecylic acid, palmitic acid, palmitoleic acid,
margaric acid, stearic acid, oleic acid, n9 t elaidic acid,
linoleic acid, linolenic acid, arachidic acid, behenic acid,
brassic acid, lignoceric acid, and nervonic acid. Meth-
ods used to determine β -carotene content in eggs were
determined using AOAC 958.05 (1990) color of egg
yolk. Egg cholesterol was determined using methods
described by Zhang et al. (1999). Egg fat hydrolysis
methods were determined using AOAC method 954.02
(1990), and fatty acid profile methods were determined
using AOCS Ce 2–66/Ce 1e 91 methods.
Statistical Analysis-Fatty Dietary
Treatments
Biochemical components of the 2 diets were measured
in triplicate and then compared using a t-test in SAS
software version 9.4 (PROC TTEST). The diets were
investigated for equal variance: if variances were de-
tected to be significantly different, the Satherwaithe
t-test P-value was reported and if variances were not
statistically significant, the pooled t-test P-value was
reported for all biochemical components.
Statistical Analysis-Hen Performance
Performance of the laying hens was measured in
number of total eggs produced, feed intake, feed con-
version ratio, and body weight for each animal over
10 wk. Analysis of variance was performed on each phe-
notype using a general linear mixed model mixed model
(PROC GLIMMIX in SAS software version 9.4). In the
first analysis room, treatment, week, and their inter-
actions were included as fixed effects. Animals were
treated as replications for diet treatments, measure-
ments taken on individual eggs were treated as subsam-
ples for each animal, and weekly measurements were
repeated measures for each animal. Random effects in-
cluded animals within treatment and room and week by
animal within treatment and room with residual error
representing subsampling error or variability between
eggs for each animal. In the second analysis, week was
treated as a trend with other factors treated same as
initial analysis. For all models, residuals were plotted
to verify that residuals were independent and normally
distributed. Means were separated at P = 0.05 using
least squares means with Tukey-Kramer adjustment for
multiple comparisons.
Statistical Analysis-Egg Quality
One egg per animal per week for 10 wk was used
for egg quality measurements including number of
meat spots and number of blood spots, egg weight
(grams), albumen height, HU, and yolk color measured
by Roche. Confidence intervals were evaluated between
treatment groups for number of meat spots and number
of blood spots.
Statistical Analysis-Egg Chemistry, Lipid
and Fatty Acid Analysis
β -Carotene levels and full lipid and fatty acid profiles
were run on a single egg from each animal measured on
weeks 1, 5, and 10. A separate egg was utilized for the β -
carotene measurement and the full lipid and fatty acid
profiles. These measurements were similarly treated as
hen performance above with the initial analysis utilizing
the week as classification effect. Since there were just 3
weekly measurements, a trend was not used to model
the week effect. Total fats, oleic acid, linoleic acid, and
the oleic-to-linoleic acid (O/L) ratio were measured on
weeks 2, 3, 4, 6, 7, 8, and 9 for eggs from all animals.
These data were initially treated with week as a class,
and then analyzed for a trend if a significant interaction
was observed.
RESULTS AND DISCUSSION
Feed Analysis
Although both experimental dietary treatments were
formulated to be isocaloric and isonitrogenous, the
high-oleic peanut + corn diet had significantly greater
amounts of lipid compared to the conventional soy-
bean meal diet, with 9.8 and 6.9% lipid, respectively
(P = 0.006, Table 1). Therefore, the corn within the
soybean meal + corn diet provided the predominant
source of energy, whereas the lipids from the high-
oleic peanuts in combination with the corn within
the high-oleic + corn diet provided dietary energy
for egg-producing layer hens. Nevertheless, the per-
centages of nitrogen and protein were similar be-
tween both experimental dietary treatment groups
(Table 1).
The soybean meal + corn experimental diet had a
significantly greater (P-value < 0.0001) content of sat-
urated fatty acids (26.4%) relative to the saturated
fatty acid content in the high-oleic peanut + corn diet
(14.3%), with the following saturated fatty acids con-
tent being significantly greater (P < 0.0001) in the con-
ventional soybean meal + corn diet (Table 1): myris-
tic acid (14:0), palmitic acid (16:0), and stearic acid
(18:0). At small quantities, arachiadic acid (20:0), be-
henic (22:0), lignoceric acid (24:0), and cerotic acid
(26:0) were the predominant saturated fatty acids found
within the high-oleic peanut + corn experimental diet
(Table 1).
The high-oleic peanut + corn diet had significantly
greater (P < 0.0001) amounts of monounsaturated
fatty acids in comparison to the soybean meal +
corn diet, with a oleic acid (18:1) content of 72.18
± 0.18 and 38.04 ± 0.19, respectively (Table 1).
Although gondoic acid (20:1) and erucic acid (22:1)
 EGGS FROM LAYER HENS FED HIGH-OLEIC PEANUTS 1737
Table 1. Chemical lipid and fatty acid analysis of dietary treatments.1

Soybean + corn diet (SBM) High-oleic peanut + corn diet (HO-PN) P-value2

% Lipid 6.88 ± 0.02 9.79 ± 0.39 0.006

% Nitrogen 2.78 ± 0.27 2.76 ± 0.04 0.90
% Protein 17.4 ± 1.7 17.2 ± 0.3 0.90
Myristic acid (14:0) 0.446 ± 0.010 0 ± 0.00 < 0.0001
Myristoleic acid (14:1cis) 0.126 ± 0.010 0 ± 0.00 0.003
Palmitic acid (16:0) 21.4 ± 0.1 7.32 ± 0.08 < 0.0001
Palmitoleic acid (16:1cis) 4.43 ± 0.17 0.085 ± 0.00 < 0.0001
Margaric acid (17:0) 0.072 ± 0.120 0 ± 0.00 0.42
Stearic acid (18:0) 4.83 ± 0.05 2.54 ± 0.02 < 0.0001
Oleic acid (18:1) 38.0 ± 0.2 72.2 ± 0.2 < 0.0001
Linoleic acid (18:2) 28.3 ± 0.2 11.3 ± 0.2 < 0.0001
α -Linolenic acid (18:3cis) 1.24 ± 0.02 0.34 ± 0.00 0.0002
Arachidic acid (20:0) 0.165 ± 0.040 1.07 ± 0.01 < 0.0001
Gondoic acid (20:1cis) 0.314 ± 0.030 1.38 ± 0.04 < 0.0001
Behenic acid (22:0) 0 ± 0.00 2.05 ± 0.03 < 0.0001
Erucic acid (22:1cis) 0 ± 0.00 0.85 ± 0.07 0.18
Lignoceric acid (24:0) 0 ± 0.00 1.25 ± 0.02 < 0.0001
Cerotic acid (26:0) 0 ± 0.00 0.40 ± 0.29 0.14
Oleic/linoleic ratio (O/L) 1.34 ± 0.01 6.40 ± 0.10 < 0.0001
Saturated fat (%) 26.4 ± 0.1 14.3 ± 0.07 < 0.0001
P/S 1.07 ± 0.01 0.79 ± 0.01 < 0.0001
1
Three replicate samples were collected from each dietary treatment and were analyzed for lipid, nitrogen, protein,
lipid, and fatty acid content. Each value represents the mean ± the standard error for each sample. Fatty acid content
(g/100 g sample).
2
Measurements were tested for equal variance. When variances were detected to be statistically significant Satherwaithe
P-values were reported, else pooled P-values were reported.

monounsaturated fatty acids were found in greater
content in the high-oleic peanut + corn diet in
comparison to the soybean meal + corn diet, they
were in small quantities (Table 1). Monounsaturated
fatty acids myristoleic acid (14:0) and palmitoleic
acid (16:1) were found to be in greater quantities in
the soybean meal + corn diet (0.126 ± 0.01, 4.43 ±
0.17) relative to the high-oleic peanut + corn diet
(0.0, 0.085 ± 0.07), respectively (Table 1).
The soybean meal + corn diet had significantly
greater amounts of (P < 0.0001, Table 1) the PUFA,
linoleic acid (28.29 ± 0.160) in comparison to the high-
oleic peanut + corn diet (11.28 ±0.200). Additionally
the soybean meal + corn diet had significantly greater
contents of omega 3 fatty acid (linolenic acid) in com-
parison to the high-oleic peanut + corn diet (P =
0.0002, Table 1).
Normal-oleic peanuts have an O/L of approximately
1.5 to 1.0 (Chamberlin et al., 2014), whereas high-oleic
peanuts have an O/L ≥ 9.0 (Davis et al., 2016, 2017).
The calculated O/L ratio in the high-oleic peanut +
corn diet and soybean meal + corn diets was 6.40 ±
0.10 and 1.344 ± 0.01, respectively (Table 1). Com-
monly, 95 out of 100 peanut kernels must pass the min-
imal fatty acid chemistry profile threshold of ≥74%
oleic acid and ≤8% linoleic acid to be classified as a
high-oleic peanut lot (Sweigart et al., 2011). Research
has demonstrated that high-oleic peanut lots may not
be meeting this threshold due to either contamina-
tion with conventional normal-oleic peanuts (Andersen
et al., 1998; Sweigart et al., 2011) and/or the presence
of immature peanut kernels within the lot that has yet
to fully express the high-oleic oil chemistry of a ma-
ture kernel (Pattee et al., 1974; Singkham et al., 2010).
Consequently, with kernel-to-kernel sampling within a
lot of high-oleic peanuts there will be a natural distri-
bution of O/L oil content (Davis et al., 2017), which
may explain oleic acid values of 72.18 ± 0.180, linoleic
acid values of 11.28 ± 0.20, and an O/L value of 6.40
± 0.10 for oil chemistry of the high-oleic peanut + corn
diet analyzed.
Layer Hen Performance
There were no significant differences in the body
weights of layer hens between the experimental treat-
ment groups at any of the time points measured (week
1 to 10, Table 2). However, there was a significant
week-to-week (wk) and treatment × week (trmt × wk)
effect on body weights of the layer hens from both
treatment groups, P = 0.029 and P = 0.0005, respec-
tively (Table 2). This indicates a trend associated with
the HO peanut diet for a body weight loss, whereas the
control group had a weight gain over the same period.
Feed conversion ratio was calculated as the ratio of
egg mass (grams) to feed weight (grams) to determine
the amount of feed utilized for egg production. Thus,
layer hens with greater feed conversion ratios were
more efficient with the utilization of feed for shell
egg production.
Although there were no overall effect on feed con-
version (P = 0.077), feed conversion was affected at
biweekly experimental time points (4, 6, 8, and 10 wk)
for hens fed the control soybean meal diet in compar-
ison to hens fed the high-oleic peanut diet (Table 2).
Although the feed conversion at these time points
1738 TOOMER ET AL.

Table 2. Performance of layer hens fed a diet with high-oleic peanuts (HO-PN) or a conventional diet with soybean meal (SBM).1

A. Body weight (g) Feed intake(g feed/bird/day) Feed conversion ratio (g egg/g feed)
SBM HOPN P-value SBM HO-PN P-value SBM HOPN P-value

Week 1 1,464 ± 37 1469 ± 37 0.93 154 ± 3 155 ± 3 0.69 na na na

Week 2 1,462 ± 37 1451 ± 37 0.83 112 ± 3 104 ± 3 0.08 na na na
Week 3 1,460 ± 37 1445 ± 37 0.78 87 ± 3 78 ± 3 0.05 0.568 ± 0.018 0.589 ± 0.018 0.42
Week 4 1,477 ± 37 1429 ± 37 0.37 96 ± 3 96 ± 3 0.90 0.600 ± 0.018 0.536 ± 0.018 0.01
Week 5 1,464 ± 37 1436 ± 37 0.59 91 ± 3 84 ± 3 0.12 0.630 ± 0.018 0.600 ± 0.018 0.24
Week 6 1,461 ± 37 1409 ± 37 0.33 107 ± 3 113 ± 3 0.16 0.530 ± 0.018 0.452 ± 0.018 0.003
Week 7 1,469 ± 37 1429 ± 37 0.45 102 ± 3 105 ± 3 0.65 0.491 ± 0.018 0.468 ± 0.018 0.36
Week 8 1,485 ± 37 1437 ± 37 0.37 113 ± 3 113 ± 3 1.00 0.554 ± 0.019 0.487 ± 0.018 0.01
Week 9 1,495 ± 37 1421 ± 37 0.17 110 ± 3 109 ± 3 0.86 0.522 ± 0.018 0.475 ± 0.018 0.06
Week 10 1,505 ± 37 1435 ± 37 0.19 107 ± 3 113 ± 3 0.21 0.533 ± 0.018 0.466 ± 0.018 0.01

B. Body weight Feed intake Feed conversion ratio

F-test P-value F-test P-value F-test P-value

Treatment (Trmt) 0.63 0.43 < 0.0001 0.97 5.65 0.02

Room (Rm) 0.64 0.43 1.30 0.26 0.71 0.40
Trmt × Rm < 0.0001 0.98 0.04 0.84 1.33 0.25
Week (Wk) 2.03 0.03 128.97 < 0.0001 31.2 < 0.0001
Trmt × Wk 3.24 0.0005 3.24 0.0005 2.64 0.01
Rm × Wk 1.13 0.34 1.98 0.03 1.99 0.05
Trmt × Rm × Wk 0.61 0.81 0.76 0.67 1.29 0.25
Trend 26.93 < 0.0001 3.31 0.07 3.14 0.08
1
Layers (24 per dietary treatment) were fed a HO-PN or SBM diet for 10 wk. (A) Body weight, feed intake, and feed conversion ratio. Reported
mean ± standard error; na = (not applicable). Feed conversion could not be calculated because ample time was needed for hens to be in full egg
production. Trend test was used to compare the slopes (variable vs. time) for hens fed HOPN or SBM diets. (B) Treatment and interaction statistics.

was lower in the HO PM, the values were close in
numerical value (Table 2). Thus, in the model, there
were significant treatment effects (P = 0.022), week ef-
fects (P < 0.0001), treatment × week effects (trt × wk)
P = 0.008). However, there were no significant differ-
ences in feed conversion between the treatment groups
at week 3, 5, 7, and 9 (Table 2).
Egg Grading, Quality and Production
All eggs produced at all time points (week 1 to 10)
between both treatment groups were graded as USDA
Grade AA of superior quality, with thick, firm egg
whites and defect-free egg yolks. Additionally, the shells
were clean and without defects. There were minimal
number of blood spots or number of meat spots, and
there was no statistical difference at the 95% confidence
interval between eggs produced from both feeding treat-
ment groups (data not shown).
The most widely used measurement of albumen (egg
white) quality is the Haugh unit, by Raymond Haugh
in 1937 (Stadelmann et al., 1995). This method con-
sists of measuring the height and thickness of the al-
bumen. Hence, fresher, higher quality eggs have thicker
egg whites and thus higher HU values. There were no
significant differences in the egg HU (Table 3), number
of eggs produced (Table 3), or the egg albumen height
(Table 4) between the treatment groups at any of the
time points measured (week 1 to 10), whereas there
were significant week effects on these 3 variables (num-
ber of eggs P < 0.0001, HU P < 0.0001, albumen height
P < 0.0001). Moreover, there was a significant trmt ×
rm effect on egg albumen height (P = 0.001, Table 4).
The yolk color Roche score was examined weekly
to determine the yolk color intensity of eggs produced
from both dietary treatments at all time points
measured (Table 4). The yolk color Roche value was
significantly greater at all time points (P < 0.0001)
measured in eggs produced from layer hens fed the
high-oleic peanut + corn diet in comparison to eggs
produced from layer hens fed the soybean meal + corn
diet (Table 4), indicating a more intense yellow-orange
colored yolk in these eggs, with main effects of treat-
ment (P < 0.0001). Additionally, there were significant
week (P < 0.0001) and trmt × wk (P < 0.0001) effects
on the yolk color Roche score, in which the Roche score
diminished weekly particularly in eggs produced from
layer hens fed the conventional soybean meal + corn
diet. Moreover, there were significant rm × wk (P <
0.0001) effects on yolk color Roche, particularly only in
eggs produced from layer hens fed the high-oleic peanut
+ corn diet. In contrast, Pesti et al. (2003) reported
that yolk color in eggs produced from layer hens fed
peanut meal to be slight and not noticeable upon
casual observation. However, in this study there were
clearly observable differences in the yolk color in eggs
produced from layer hens fed the high-oleic peanut diet
in comparison to eggs produced from layer hens fed the
soybean meal diet.
Eggs produced from layer hens fed the soybean meal
+ corn diet had egg weights that were significantly
greater in size than eggs from layer hens fed the high-
oleic peanut + corn diet at all time points measured
(week 1 to 10), with significant treatment (P = 0.0001)
and week (P = 0.003) effects on egg weights (Table
3). Interestingly, the total number of eggs produced
 EGGS FROM LAYER HENS FED HIGH-OLEIC PEANUTS 1739
Table 3. Egg production parameters and quality of eggs produced by layer hens fed a diet with high-oleic peanuts (HO-PN) or a
conventional diet with soybean meal (SBM).1

A. Number of eggs produced Egg weight (g) Haugh unit (HU)

SBM HO-PN P-value SBM HO-PN P-value SBM HO-PN P-value

Week 1 5.58 ± 0.24 5.71 ± 0.24 0.72 58.5 ± 0.7 55.8 ± 0.7 0.01 92.6 ± 0.8 93.1 ± 0.8 0.67
Week 2 7.46 ± 0.24 7.25 ± 0.24 0.55 60.3 ± 0.7 56.5 ± 0.7 < 0.0001 95.2 ± 0.8 94.3 ± 0.8 0.45
Week 3 5.67 ± 0.24 5.75 ± 0.24 0.81 59.0 ± 0.7 55.9 ± 0.7 0.003 92.8 ± 0.8 93.4 ± 0.8 0.60
Week 4 6.42 ± 0.24 6.29 ± 0.24 0.72 58.7 ± 0.7 54.8 ± 0.7 < 0.0001 94.6 ± 0.9 94.8 ± 0.9 0.92
Week 5 6.50 ± 0.24 6.25 ± 0.24 0.47 58.5 ± 0.7 55.8 ± 0.7 0.009 94.4 ± 0.8 94.3 ± 0.8 0.98
Week 6 6.38 ± 0.24 6.21 ± 0.24 0.06 59.8 ± 0.7 56.3 ± 0.7 0.001 94.1 ± 0.8 95.6 ± 0.8 0.21
Week 7 6.00 ± 0.24 6.04 ± 0.24 0.90 59.5 ± 0.7 56.5 ± 0.7 0.004 92.2 ± 0.8 92.0 ± 0.8 0.88
Week 8 6.79 ± 0.24 6.63 ± 0.24 0.63 59.3 ± 0.7 56.2 ± 0.7 0.003 92.7 ± 0.9 92.2 ± 0.9 0.65
Week 9 6.38 ± 0.24 6.21 ± 0.24 0.63 59.5 ± 0.7 57.1 ± 0.7 0.02 88.1 ± 0.8 87.9 ± 0.8 0.86
Week 10 6.38 ± 0.24 6.29 ± 0.24 0.81 59.5 ± 0.7 57.1 ± 0.7 0.02 88.1 ± 0.8 87.9 ± 0.8 0.86

B. # Eggs produced (#) Egg weight (g) Haugh unit (HU)

F-test P-value F-test P-value F-test P-value

Treatment (Trmt) 0.09 0.77 14.30 0.0001 0.01 0.94

Room (Rm) 0.15 0.70 1.00 0.32 1.19 0.28
Trmt × Rm 0.47 0.50 0.09 0.76 11.37 0.002
Week (Wk) 24.10 < 0.0001 2.89 0.003 40.97 < 0.0001
Trmt × Wk 0.35 0.97 0.65 0.75 0.66 0.74
Rm × Wk 1.70 0.08 0.43 0.92 1.08 0.38
Trmt × Rm × Wk 0.40 0.95 0.69 0.72 1.22 0.28
1
Twenty-four animals per dietary treatment were fed either an HO-PN or SBM diet for 10 wk. (A) Egg data. Egg Haugh unit (HU) was calculated
to determine egg albumen quality. Reported mean ± standard error. (B) Treatment and interaction statistics.

Table 4. Egg quality and β -carotene levels of eggs produced by layer hens fed a diet of high-oleic peanuts (HO-PN) or a conventional
diet with soybean meal (SBM).1

A. Albumen height2 (mm) Yolk color3 (Roche score 1 to 15) β -Carotene4 (ppm)
SBM HO-PN P-value SBM HOPPN P-value SBM HO-PN P-value

Week 1 8.60 ± 0.16 8.54 ± 0.16 0.78 5.09 ± 0.15 6.63 ± 0.15 < 0.0001 11.44 ± 0.35 12.21 ± 0.35 0.12
Week 2 9.18 ± 0.16 8.81 ± 0.15 0.10 5.74 ± 0.15 6.97 ± 0.15 < 0.0001 na na na
Week 3 8.62 ± 0.16 8.60 ± 0.16 0.92 4.67 ± 0.15 6.64 ± 0.15 < 0.0001 na na na
Week 4 9.01 ± 0.16 8.82 ± 0.16 0.43 4.18 ± 0.15 6.14 ± 0.16 < 0.0001 na na na
Week 5 8.93 ± 0.16 8.79 ± 0.16 0.53 4.48 ± 0.15 6.48 ± 0.15 < 0.0001 7.74 ± 0.35 12.75 ± 0.39 < .0001
Week 6 8.92 ± 0.16 9.09 ± 0.16 0.43 4.11 ± 0.15 6.49 ± 0.15 < 0.0001 na na na
Week 7 8.55 ± 0.16 8.35 ± 0.16 0.38 3.76 ± 0.15 5.62 ± 0.15 < 0.0001 na na na
Week 8 8.64 ± 0.16 8.39 ± 0.16 0.28 3.72 ± 0.16 6.10 ± 0.16 < 0.0001 na na na
Week 9 7.82 ± 0.16 7.64 ± 0.15 0.41 2.87 ± 0.15 5.96 ± 0.15 < 0.0001 na na na
Week 10 7.82 ± 0.16 7.64 ± 0.15 0.41 2.87 ± 0.15 5.96 ± 0.15 < 0.0001 8.27 ± 0.26 12.40 ± 0.36 < .0001

B. Albumen height2 Yolk color3 β -Carotene4

F-test P-value F-test P-value F-test P-value

Treatment (Trmt) 0.71 0.40 301.00 < 0.0001 128.06 < 0.0001
Room (Rm) 0.60 0.44 1.22 0.28 2.69 0.11
Trmt × Rm 11.60 0.001 4.12 0.05 1.58 0.22
Week (Wk) 38.20 < 0.0001 49.60 < 0.0001 45.90 < 0.0001
Trmt × Wk 0.89 0.53 11.16 < 0.0001 55.09 < 0.0001
Rm × Wk 1.05 0.40 5.43 < 0.0001 6.80 0.002
Trmt × Rm × Wk 1.04 0.40 1.47 0.16 9.42 0.0002
Trend na na 60.40 < 0.0001 128.06 na
1
Twenty-four animals per dietary treatment were fed either an HO-PN or SBM diet for 10 wk. (A) Egg data. Each value represents the mean ±
the standard error. Trend test compared the slopes (variable vs. time) for the 2 diets.
2
Egg albumen height (mm) was calculated to determine egg albumen quality.
3
Yolk color was determined using the Roche Color Fan color index 1 to 15 to distinguish the yolk color from lightest to darkest color intensity.
4
Eggs collected weeks 1, 5, and 10 were analyzed for β -carotene content; testing at other time points was cost prohibitive and reported as na =
not applicable. (B) Treatment and interaction statistics.

from layer hens fed the high-oleic peanut + corn diet
categorized by weight (United States Department of
Agriculture egg classification system) was 60% medium
size eggs, 35% large size eggs, 3% extra-large size eggs,
and 2% small size eggs. The total number of eggs pro-
duced from layer hens fed the soybean meal + corn diet
categorized by weight (United States Department of
Agriculture egg classification system) was 66% large
size eggs, 24% medium size eggs, 10% extra-large size
eggs, and 0.3% small size eggs. These results are similar
to results reported by Van Elswyk et al. (1994) demon-
strating that layer hens fed unsaturated fatty acids in
1740 TOOMER ET AL.

the diet produced reduced size eggs in response to the

Twenty-four animals per dietary treatment were fed either an HO-PN or SBM diet for 10 wk. (A) Egg data. Eggs collected at 1, 5, and10 wk were analyzed by ATC Scientific for lipid and fatty
< 0.0001

< 0.0001
< 0.0001

< 0.0001
< 0.0001

< 0.0001
0.55
0.56

0.19
0.51
menhaden (fish) oil feeding in comparison to the con-

Total oleic acid3

P-value
P
trols. This attribute could be of potential benefit within

Total oleic acid3 (18:1)

the commercial egg production industry by reducing the
number of oversized eggs produced in older laying hens.

48.2 ± 0.5
53.1 ± 0.5
56.0 ± 0.5

653.36
0.37
0.34
90.25
43.04
1.69
0.69
Table 5. Lipid and fatty acid content of eggs produced by layer hens fed a diet with high-oleic peanuts (HO-PN) or a conventional diet with soybean meal (SBM).1

HO-PN

F-test
Layer hens typically start to lay eggs around 5 mo
(20 to 21 wk) of age and continue to lay for 12 mo to
52 wk (FAO, 2003). Birds typically begin producing

39.5 ± 0.5
38.2 ± 0.5
41.6 ± 0.5
eggs in their 20th or 21st week and continue for slightly

SBM
over a year (52 wk), with eggs tending to increase in
size until the end of the egg production cycle (Silver-

< 0.0001

< 0.0001
< 0.0001
< 0.0001

< 0.0001
sides and Scott, 2001; Padhi et al., 2013). Egg size has

0.860
Total palmitoleic acid3

0.62

0.73
0.77

0.14
been shown to be influenced by genetic factors (breed,

P-value
P
Total palmitoleic acid3 (16:1)
breeding system, body weight, and age), nutritional fac-
tors (energy and feed intake, protein, lipids, feeding

1.98 ± 0.06
1.52 ± 0.06
1.52 ± 0.06
program), and pullet management (lighting program,

643.26
0.25

0.32
0.09

61.32
2.00
0.16
HO-PN

F-test
stress, and disease management). For example, egg size
is directly influenced by body weight; in that for ev-
ery 45 g of body weight increase from 18 wk of age,

2.88 ± 0.06
3.29 ± 0.06
3.37 ± 0.06
there is a 0.5-g increase in egg size (Duraisamy, 2011).

SBM
Therefore, as the layer hen ages in the production cycle
and increases in body weight there is a proportionate
increase in egg size (Duraisamy, 2011). Thus, a feeding

< 0.0001

< 0.0001

< 0.0001
< 0.0001
< 0.0001

0.110
0.86
0.32

0.22
0.27
program, which aims to reduce egg size, is of significant

Total stearic acid3

P-value
P
interest in the commercial egg production industry as
a mechanism to manage egg size with increasing pro- Total stearic acid3 (18:0)
duction age of layer hens. In conclusion, the only reduc-
7.37 ± 0.13
5.91 ± 0.14
6.11 ± 0.13

46.60
0.03
1.01
116.20
1.54
1.33
2.29
HO-PN
tion in performance due to feeding layer hens’ high-oleic

F-test
peanut + corn diet was smaller egg weights over the
10-wk trial, while all other production parameters were
8.80 ± 0.13
7.67 ± 0.13
8.05 ± 0.13

similar. In future studies, we aim to examine the ef-

SBM

fects of a high-oleic peanut diet on egg size in late-stage

egg production.
< 0.0001

< 0.0001

< 0.0001
< 0.0001

< 0.0001
< 0.0001

0.90
0.31

0.33
0.99
Total palmitic Acid3
P-value
P

Egg β-Carotene, Lipid and Fatty Acid

Total palmitic acid3 (16:0)

Content
23.1 ± 0.2
21.4 ± 0.2
21.0 ± 0.2

0.01
1.05
28.88
10.68
1.12
0.01
HO-PN

Similar to the egg yolk color, β -carotene levels were

74.8

Units of g/100 g sample. (B) Treatment and interaction statistics.

F-test

significantly greater in eggs produced from layer hens

acid content. Each value represents the mean ± standard error.

fed the high-oleic peanut + corn diet in comparison to

25.9 ± 0.2
26.9 ± 0.2
26.9 ± 0.2

eggs produced from layer hens fed the soybean meal +

SBM

corn diet at week 5 (P < 0.0001, Table 4) and week 10

(P < 0.0001, Table 4). Hence, implying the β -carotene
content in the eggs produced was influenced by the
< 0.0001

< 0.0001
< 0.0001
0.35

0.06
0.08
0.01

0.10
0.01
0.82

lipid/fatty acid composition and/or polyphenolic com-

Total choleserol2
P-value
P

ponents of the high-oleic peanuts.

Total choleserol2

β -Carotene is a carotenoid found in plants that gives

10.50
0.34
0.08
7.38
2.38
4.79
0.19

plants rich yellow and deep orange hues and is a pre-

385 ± 6
373 ± 7
392 ± 6
HO-PN

F-test

cursor of vitamin A (retinol). Today, there is a grow-

Units of mg/100 g sample.

ing body of scientific literature, which indicates the

health benefits of β -carotene and other carotenoids in
377 ± 6
349 ± 6
360 ± 6
SBM

the prevention of chronic diseases in humans (Wood-

side et al., 2015; Zaheer, 2017). Carotenoids represent
Trmt × Rm × Wk

a group of lipid soluble antioxidant-rich bioactive com-

pounds present in plants. Conventional commercially
Room (Rm)
Trmt × Rm

Trmt × Wk
Week (Wk)

Rm × Wk

produced eggs yolks are rich in carotenoids, lutein,

Week 10
Week 1
Week 5

and zeaxanthin (Zaheer, 2017). However, lutein and

Trmt

2
3
A.

B.

zeaxanthin content within the egg is highly subject to

 EGGS FROM LAYER HENS FED HIGH-OLEIC PEANUTS 1741
oxidation during, egg processing, storage, and transport

Total linoleic acid (18:2)

< 0.0001
< 0.0001
< 0.0001

< 0.0001

< 0.0001
< 0.0001

Twenty-four animals per dietary treatment were fed either an HO-PN or SBM diet for 10 wk. (A) Egg data (units g/100 g sample). Eggs collected at 1, 5, and 10 wk were analyzed by ATC
P-value

P-value

0.29

0.40
0.77

0.25
and/or cooking (Zaheer, 2017).
Studies by Olson et al. (2008) demonstrated that

Total linoleic acid (18:2)

carotenoid pigments found naturally within plant-based

1.11 ± 0.02
0.84 ± 0.03
0.84 ± 0.03
feedstuff of layer hen diets are transferred to the egg

HO-PN

150.23
1.13

0.94
0.09
202.07
13.90

1.41
F-test
yolk of the eggs produced. Egg yolk color has been
shown to also be affected by several different compo-
nents within the diet, such as lipid profile (Suksombat

Table 6. Fatty acid content of eggs produced by layer hens fed a diet with high-oleic peanuts (HO-PN) or a conventional diet with soybean meal (SBM).1
et al., 2006) and type and concentration of carotenoids

1.40 ± 0.03
1.24 ± 0.03
1.05 ± 0.03
(Leeson and Caston, 2004; Karadas et al., 2006). Inter-

SBM
estingly, studies by Chung et al. (2004) revealed that
the carotenoid lutein was more bioavailable from en-
riched eggs in comparison to lutein found in spinach and
dietary supplements. This research also demonstrated

< 0.0001
< 0.0001
< 0.0001

< 0.0001

< 0.0001
< 0.0001
P-value

P-value

0.13

0.22
0.80

0.16
improved intestinal absorption of lutein when consumed

acids
with dietary lipids, suggesting that eggs may be supe-

Omega 6 fatty
Omega 6 fatty acids
rior delivery system for some carotenoids (Chung et al.,

1.26 ± 0.03
0.93 ± 0.03
1.00 ± 0.03
2004).

HO-PN

6.72
101.12
2.39

107.16

1.55
0.07

1.87
F-test
Karadas et al. (2006) demonstrated that supplement-
ing the diets of female quail with natural carotenoids

Scientific for lipid and fatty acid content. Each value represents the mean ± standard error. (B) Treatment and interaction statistics.
(alfafa concentrated, tomato powder, and/or marigold
extract) enriched egg yolks with carotenoids lutein,

1.59 ± 0.03
1.31 ± 0.03
1.19 ± 0.03
zeaxanthin, lycopene, and β -carotene. This work paral-

SBM
lels earlier research by Jiang et al. (1994), which demon-
strated that dietary supplementation of vitamin E and
β -carotene in the diets of layer hens enriched egg yolk
proportionately with vitamin E and β -carotene levels

< 0.0001

< 0.0001

< 0.0001

< 0.0001
P-value

P-value
0.09

0.09
0.07
0.54

0.01
0.41
in comparison to the controls. However, these feeding

Omega 3 fatty acids

regimens are not commercially viable due to the asso-
ciated cost of the inclusion of specialty feed ingredients
Omega 3 fatty acids

0.013 ± 0.001
0.004 ± 0.002
(dietary carotenoid supplements, alfalfa, tomato pow- 0.007 ± 0.002
HO-PN

F-test
26.26

38.10
2.54
3.33
0.39

4.58
0.91
der, marigold) in the diets of layer hens to enrich the
eggs (Jiang et al., 1994). In general, this study implies
that feeding layer hens’ high-oleic peanuts may be an
economical and effective means to enrich egg nutrition
0.023 ± 0.002
0.007 ± 0.002
0.014 ± 0.002

and yolk color and may be of value to peanut and egg

producers globally.
SBM

Today, consumer demand for food products of supe-

rior quality and nutrition has led to increased interest
and research in livestock feeding studies with aims to
enrich and/or modify the lipid and nutrient profile of
< 0.0001
< 0.0001

< 0.0001

< 0.0001
< 0.0001
P-value

P-value
0.71

0.69
0.27

0.01
0.34
n9 elaidic acid (18:1)

the eggs and meat produced. Although current feeding

studies have demonstrated the health benefits of the
n9 elaidic acid (18:1)

moderate consumption of high-oleic peanuts in the diet

4.14 ± 0.31
6.29 ± 0.36
4.59 ± 0.33

(Moreira Alves et al., 2014, Alves et al., 2014; Moreira

HO-PN

F-test
39.59

50.58
0.16
1.24

8.33
4.51
1.10

et al., 2016; Barbour et al., 2017), there are no pre-

vious studies which have examined the use, economic,
and health advantages of utilizing high-oleic peanuts
as a feed ingredient to enrich eggs and meat produced
4.30 ± 0.32
8.78 ± 0.32
6.99 ± 0.32

with heart healthy monounsaturated fatty acids. Earlier

SBM

feeding trials revealed that the fatty acid composition

of poultry meat (Marion and Woodroof, 1963; Hulan
et al., 1988; Yau et al., 1991) and egg yolk (Cruick-
Trmt × Rm × Wk

shank, 1934; Sell et al., 1968; Ohtake and Hoshino,

1976; El-Hussainy et al., 1983) is readily altered by
Room (Rm)
Trmt × Rm

Trmt × Wk
Week (Wk)

dietary manipulation of fatty acids. Therefore, in this

Rm × Wk
Week 10

study we aimed to determine the effects of feeding

Week 1
Week 5

Trmt

layer hens’ high-oleic peanut diet on the fatty acid and

1
A.

B.
1742 TOOMER ET AL.

Table 7. Total linoleic and oleic fatty acid content of eggs produced by layer hens fed a diet with high-oleic peanuts (HO-PN) or a
conventional diet with soybean meal (SBM).1

A. Total linoleic acid (18:2) (%) Total oleic acid (18:1) (%) Oleic: linoleic ratio
SBM HO-PN P-value SBM HO-PN P-value SBM HO-PN P-value

Week 2 13.5 ± 0.4 9.5± 0.4 < 0.0001 36.9 ± 0.6 49.6 ± 0.5 < 0.0001 2.76 ± 0.09 5.28 ± 0.09 < 0.0001
Week 3 12.6 ± 0.4 10.3 ± 0.4 < 0.0001 36.6 ± 0.6 38.9 ± 0.6 0.004 3.03 ± 0.09 3.80 ± 0.09 < 0.0001
Week 4 14.0 ± 0.4 10.4 ± 0.4 < 0.0001 28.2 ± 0.5 37.1 ± 0.5 < 0.0001 2.03 ± 0.09 3.59 ± 0.09 < 0.0001
Week 6 13.1 ± 0.4 10.4 ± 00.4 < 0.0001 29.7 ± 0.5 37.2 ± 0.5 < 0.0001 2.28 ± 0.09 3.60 ± 0.09 < 0.0001
Week 7 13.4 ± 0.4 10.0 ± 0.4 < 0.0001 29.5 ± 0.6 38.9 ± 0.5 < 0.0001 2.21 ± 0.09 3.89 ± 0.09 < 0.0001
Week 8 13.3 ± 0.4 11.1 ± 0.40 < 0.0001 31.5 ± 0.6 40.7 ± 0.5 < 0.0001 2.38 ± 0.09 4.21 ± 0.09 < 0.0001
Week 9 12.7 ± 0.4 10.0 ± 0.37 < 0.0001 31.5 ± 0.6 40.8 ± 0.5 < 0.0001 2.49 ± 0.09 4.22 ± 0.09 < 0.0001

B. Total linoleic acid (18:2) Total oleic acid (18:1) Oleic: linoleic ratio
F-test P-value F-test P-value F-test P-value

Treatment (Trmt) 165.10 < 0.0001 541.07 < 0.0001 672.75 < 0.0001
Room (Rm) 0.29 0.59 0.94 0.34 0.03 0.86
Trmt × Rm 0.45 0.51 1.36 0.25 0.14 0.71
Week (Wk) 1.52 0.17 96.89 < 0.0001 44.46 < 0.0001
Trmt × Wk 1.65 0.14 18.40 < 0.0001 19.04 < 0.0001
Rm × Wk 1.34 0.24 3.59 0.002 1.58 0.16
Trmt × Rm × Wk 1.8 0.10 0.27 0.95 1.68 0.13
Trend na na 0.45 0.50 0.04 0.84
1
Twenty-four animals per dietary treatment were fed either an HO-PN or SBM diet for 10 wk. (A) Egg data. Eggs collected at weeks 2, 3, 4, 6,
7, 8, and 9 were analyzed for lipid and fatty acid content at the Market Quality & Handling Research Unit at. Each value represents the mean ±
standard error. Trend test compared the slopes (variable vs. time) of the 2 diets. (B) Treatment and interaction statistics.

lipid profile of shell eggs produced from layer hens for
10 wk.
In this study, total cholesterol from eggs produced
from layer hens fed the high-oleic peanut + corn diet
was significantly greater than the total cholesterol from
eggs produced from layer hens fed the conventional
soybean meal + corn diet at week 5 (P = 0.01) and
week 10 (P < 0.0001) experimentally (Table 5), with
significant treatment effects (P < 0.0001), wk effects
(P < 0.0001), and rm × wk effects (P = 0.010). To-
tal saturated fatty acids, palmitic acid and stearic
acid, were significantly higher in the eggs produced
from layer hens fed the conventional soybean meal +
corn diet in comparison to eggs produced from layer
hens fed the high-oleic peanut + corn diets at week 1
(P < 0.0001), week 5 (P < 0.0001), and week 10
(P < 0.0001) of the experiment (Table 5), with signifi-
cant treatment (P < 0.0001), wk (P < 0.0001), and trmt
× wk effects (P < 0.0001 palmitic acid only). Addition-
ally, trans fat n9 elaidic acid was significantly higher in
eggs produced from layer hens fed the conventional soy-
bean meal + corn diet at week 5 (P < 0.0001) and week
10 (P < 0.0001) experimentally (Table 6), with trmt
(P < 0.0001), wk (P < 0.0001), trmt × wk (P < 0.0001),
and rm × wk (P = 0.010) effects.
Eggs produced from layer hens fed the soybean
meal + corn diet had significantly higher content
of monounsaturated fatty acid, palmitoleic acid
(Table 5), and polyunsaturated fatty acid, linoleic acid
(Table 6) at week 1 (P < 0.0001), week 5 (P < 0.0001),
and week 10 (P < 0.0001) experimentally, with trmt
(P < 0.0001) and trmt × wk (P < 0.0001) effects.
Conversely, the content of monounsaturated fatty acid,
oleic acid content was significantly higher in eggs pro-
duced from layer hens fed the high-oleic peanut + corn
diet at week 1 (P < 0.0001), week 5 (P < 0.0001), and
week 10 (P < 0.0001) experimentally (Table 5), with
trmt (P < 0.0001), wk (P < 0.0001), and trmt × wk
(P < 0.0001) effects. Also, eggs produced at week 2, 3,
4, 6, 7, 8, and 9 by hens fed the high-oleic peanut + corn
diet had significantly greater total oleic acid content in
comparison to eggs produced by hens fed the soybean
meal conventional diet, with significant treatment
(P < 0.0001), wk (P < 0.0001), trmt × wk (P <
0.0001), and rm × wk (P = 0.002) effects (Table 7).
Moreover, eggs produced from layer hens fed the high-
oleic peanut + corn diet had higher O/L ratios than the
other treatment group at all the time points measured
(P < 0.0001, Table 7), with significant treatment (P <
0.0001), wk (P < 0.0001), and trmt × wk (P < 0.0001)
effects. In parallel, eggs produced from layer hens fed
the soybean meal + corn diet had significantly greater
levels of total linoleic acid in comparison to eggs pro-
duced from layer hens fed the high-oleic peanut + corn
diet at weeks 2, 3, 4, 6, 7, 8, and 9 (P < 0.0001, Table 7),
with significant treatment effects (P < 0.0001).
Lastly, chemical analysis was conducted to deter-
mine the total contents of omega 3 and omega 6 family
of PUFA. Although the levels of omega 3 and omega
6 PUFA were very low in eggs from both treatment
groups, eggs produced from layer hens fed the soy-
bean meal + corn diet had significantly greater levels
of omega 3 and omega 6 PUFA in comparison to eggs
produced from layer hens fed the high-oleic peanut +
corn diets at week 1 (omega 3 P < 0.0001, omega 6
P < 0.0001), week 5 (omega 3 P = 0.09, omega 6
 EGGS FROM LAYER HENS FED HIGH-OLEIC PEANUTS 1743
P < 0.0001), and week 10 (omega 3 P < 0.0001,

0.121 ± 0.009 0.065 ± 0.009 < 0.0001

Twenty-four animals per dietary treatment were fed either an HO-PN or SBM diet for 10 wk. (A) Egg data. Eggs collected at weeks 1, 5, and 10 were analyzed by ATC Scientific for lipid and fatty
0.29
0.69
omega 6 P < 0.0001) experimentally (Table 6). More-

P
Total n6 c,c,c gamma-linolenic
over, there were significant treatment (P < 0.0001)

0.061 ± 0.009 0.075 ± 0.010

2.8 E-17 ± 0.009 0.006 ± 0.011
and wk (P < 0.0001) effects on omega 3 and omega

Total n6 c,c,c gamma-linolenic

< 0.0001
< 0.0001
HO-PN

0.24
0.35
0.13
0.05
0.23
6 PUFA contents in eggs in both treatment groups

acid (18:3)

P-value
(Table 6).

acid (18:3)
Although the content of total tridecanoic in eggs pro-
duced from both feeding treatments was ≤ 0.031, to-

Table 8. Fatty acid content of eggs produced by layer hens fed a diet with high-oleic peanuts (HO-PN) or a conventional diet with soybean meal (SBM).1

2.27
3.93
1.48

8.17
1.44
1.07
45.27
tal tridecanoic acid content (Table 8) was significantly

SBM

F-test
greater in eggs produced from layer hens fed the soy-
bean meal + corn diet at week 5 (P < 0.0001) and
week 10 (P = 0.000), with significant treatment (P <

0.248 ± 0.015 0.124 ± 0.015 < 0.0001

1.1E-17 ± 0.006 0.025 ± 0.006 < 0.0001 0.158 ± 0.006 0.193 ± 0.006 < 0.0001 0.136 ± 0.015 0.066 ± 0.015 < 0.0001
0.0001), week (P < 0.0001), and trmt × wk (P = 0.040)

0.06
P
effects. Similarly, levels of total pentadecyclic acid and

0.007 ± 0.006 0.046 ± 0.007 < 0.0001 0.164 ± 0.006 0.196 ± 0.007 < 0.0001 0.084 ± 0.015 0.041 ± 0.017
total margaric acid in eggs of were very low. How-

Total linolenic

< 0.0001

< 0.0001
HO-PN
acid (18:3)
ever, eggs produced from layer hens fed the high-oleic

0.14
0.41

0.02
0.01
0.04
P-value
Total linolenic
peanut + corn diet had significantly greater contents

acid (18:3)
of total pentadecyclic acid and total margaric acid in
comparison to eggs produced from a traditional soy-

SBM
bean meal diet at weeks 5 and 10 (Table 8). Moreover,

34.00
2.24
0.70
37.91
3.98
4.90
1.01
F-test
there were significant treatment (P < 0.0001), room
(P = 0.030), trmt × rm (P < 0.0001), wk (P < 0.0001),
and trmt × wk (P = 0.040) effects on total pentade-

0.07
P

acid content (g/100 g sample). Each value represents the mean ± standard error. (B) Treatment and interaction statistics.
cyclic levels between treatment groups (Table 8). There

Total margaric acid (17:0)

were only significant treatment effects (P < 0.0001)

0.178 ± 0.006 0.193 ± 0.006

Total margaric acid (17:0)

on total margaric levels between the treatment groups

< 0.0001
HOPN

0.58
0.31
0.20
0.16
0.20
0.31
(Table 8).

P-value
Total linolenic acid levels were significantly greater
in eggs produced from layer hens fed the soybean meal
+ corn diet at week 1 (P < 0.0001) and week 10
SBM

24.24

1.64
1.20
0.31
1.05
1.66
1.85
(P < 0.0001) in comparison to eggs of the other

F-test
treatment group (Table 8), with significant treatment
(P = 0.0001), wk (P < 0.0001), trmt × wk (0.020), and
0.25

rm × wk (P = 0.010) effects. Nevertheless, total n6,

P

c, c, c, gamma-linolenic acid content was only signifi-

Total pentadecylic acid (15:0)

− 2.3E-18 ± 0.006 0.010 ± 0.006

cantly greater in eggs produced from layer hens fed the

Total pentadecylic acid (15:0)

< 0.0001

< 0.0001
< 0.0001
HO-PN

soybean meal + corn diet at week 1 (P < 0.0001) in

0.41
0.03

0.04
0.18
P-value

comparison to eggs produced from layer hens fed the

high-oleic peanut + corn diet (Table 8), with signifi-
cant wk (P < 0.0001) and trmt × wk (P < 0.0001)
19.16

9.72
5.04

7.56
3.32
1.75
0.90

effects.
SBM

Total homo-gamma-linolenic acid levels were signif-

F-test

icantly greater only at week 10 (P = 0.010) in eggs

produced from layer hens fed the high-oleic peanut +
0.026 ± 0.002 0.015 ± 0.002 < 0.0001
0.031 ± 0.002 0.021 ± 0.002 < 0.0001

corn diet in comparison to the other treatment group

0.16
P

(Table 9), with significant room (P < 0.0001), wk (P <

Total tridecanoic acid (13:0)

0.0001), rm × wk (P < 0.0001), and trmt × rm × wk

0.017 ± 0.002 0.013 ± 0.002

Total tridecanoic acid (13:0)

(P = 0.010) effects. Total arachidonic acid levels were

< 0.0001

< 0.0001
HO-PN

P-value

0.32

0.37
0.84

0.04
0.09

significantly greater only at week 1 in eggs produced

from layer hens fed the soybean meal + corn diet in
comparison to the other treatment group, with signif-
icant rm (P < 0.0001), trmt × rm (P = 0.020), wk
SBM

1.00
25.26

1.01
23.24
0.04

3.44
2.51

(P < 0.0001), trmt × wk (P = 0.040), and rm × wk

F-test

(P = 0.020) effects (Table 9). The remaining fatty

acids analyzed included caproic acid, caprylic acid,
Trmt × Rm× Wk

capric acid, undecanoic acid, myristoleic acid, gadoleic

Room (Rm)
Trmt × Rm

Trmt × Wk
Week (Wk)

acid, and brassic acid. The contents of these fatty acids

Rm × Wk
Week 10

were ≤ 0.04 g/100 g of feed or were undetectable (data

Week 1
Week 5

Trmt

1
A.

B.

not shown).
1744 TOOMER ET AL.

Table 9. Fatty acid content of eggs produced by layer hens fed a diet with high-oleic peanuts (HO-PN) or a conventional diet with
soybean meal (SBM).1

A. Total homo-gamma-linolenic acid (20:3) Total arachidonic acid (20:4)

SBM HO-PN P-value SBM HO-PN P-value

Week 1 0.016 ± 0.005 0.010 ± 0.005 0.47 0.152 ± 0.007 0.132 ± 0.007 0.03
Week 5 0.020 ± 0.005 0.021 ± 0.006 0.90 0.046 ± 0.007 0.061 ± 0.008 0.15
Week 10 0.091 ± 0.005 0.113 ± 0.006 0.01 1.3E-17 ± 0.007 2.7E-17 ± 0.007 1.00

B. Total homo-gamma-linolenic acid Total arachidonic acid

F-test P-value F-test P-value

Treatment (Trmt) 1.68 0.20 0.11 0.47

Room (Rm) 23.02 < 0.0001 11.87 < 0.0001
Trmt × Rm 2.00 0.16 5.57 0.020
Week (Wk) 165.27 < 0.0001 233.53 < 0.0001
Trmt × Wk 3.48 0.03 3.25 0.04
Rm × Wk 10.72 < 0.0001 3.97 0.02
Trmt × Rm × Wk 4.75 0.01 1.44 0.24
1
Twenty-four animals per dietary treatment were fed either an HO-PN or SBM diet for 10 wk. (A) Egg data. Eggs collected at weeks 1, 5, and 10
were analyzed by ATC Scientific for lipid and fatty acid content (g/100 g sample). Each value represents the mean ± standard error. (B) Treatment
and interaction statistics.

Figure 2. Immunoreactivity of protein extracts from pooled total egg samples from eggs produced by layer hens fed a diet with high-oleic
peanuts (HOPN) vs. a conventional diet with soybean meal (SBM) at week 1. Protein extracts from pooled egg samples (75 μ g per lane) were
run electrophoretically on a 10% polyacrylamide gel. To determine immunereactivity, resolved proteins were transferred to a nitrocellulose mem-
brane and immunoblotted with rabbit IgG anti-peanut agglutinin antibody (1:1000). Biodetection was determined with chromogenic peroxidase
substrate-based detection of HRP activity. (A) Immunoblotting results of total egg proteins from hen(s) #1 to 12 (HOPN) and hen(s) 13 to 24
(SBM). Extracted proteins (75 μ g) from peanut flour were utilized as a positive control. (B) Immunoblotting results of total egg proteins from
hen(s) #25 to 36 (SBM) and hen(s) 37 to 48 (HOPN). Extracted proteins (75 μ g) from peanut flour were utilized as a positive control.

Temporal Changes of Egg Fatty Acids and
Lipids
Two components enter into the influence of diet and
the rate at which the dietary change will affect the per-
formance criteria. First is hen age and second is the
nutrient composition of the diet. The age of the hen
has been shown (Al-Batshan et al., 1994) to negatively
influence the utilization of some of the nutrients in the
diet. Therefore, as diets are changed during a phase
feeding program in the layer industry understanding in-
teraction of age and the speed in which the diet change
will have an impact on the production criteria is impor-
tant. This study also examined the rate (week to week)
 EGGS FROM LAYER HENS FED HIGH-OLEIC PEANUTS 1745

Figure 3. Immunoreactivity of protein extracts from pooled total egg samples from eggs produced by layer hens fed a diet with high-oleic
peanuts (HOPN) vs. a conventional diet with soybean meal (SBM) at week 5. Protein extracts from pooled egg samples (75 μ g per lane) were
electrophoretically ran on a 10% polyacrylamide gel. To determine immunereactivity, resolved proteins were transferred to a nitrocellulose mem-
brane and immunoblotted with rabbit IgG anti-peanut agglutinin antibody (1:1000). Biodetection was determined with chromogenic peroxidase
substrate-based detection of HRP activity. (A) Immunoblotting results of total egg proteins from hen(s) #1 to 12 (HOPN) and hen(s) 13 to 24
(SBM). Extracted proteins (75 μ g) from peanut flour were utilized as a positive control. (B) Immunoblotting results of total egg proteins from
hen(s) #25 to 36 (SBM) and hen(s) 37 to 48 (HOPN). Extracted proteins (75 μ g) from peanut flour were utilized as a positive control.

at which the feeding of high-oleic peanuts had on hen
performance and the quality and nutrient composition
of the eggs they produced.
This study demonstrates that egg fatty acid and lipid
composition can be greatly influenced rapidly (approx-
imately 1 to 2 wk) with dietary modification of di-
etary fat fed to egg-producing hens. Eggs from the
high-oleic peanut feeding group had a reduction in the
content of saturated fatty acids (total palmitic and to-
tal stearic) and an increase in oleic fatty acid content
that was very apparent after 1 to 2 wk of feeding the
high-oleic peanut diet in comparison to the controls
(Table 5). Additionally, there was a significant trmt ×
wk interaction shown in Table 2 indicating that the in-
troduction of a new diet to a laying hen can have an
impact on body weight and feed intake within 7 d of the
change. However, the influence of a dietary change on
egg production characteristics does not come into play
until after approximately 4 wk as shown by the drop in
feed conversion of 0.536 g egg/g feed for the hens on the
high-oleic peanut diet, which accounts for the changes
in production parameters of feed consumption and egg
weight.
Interestingly, there was no influence of diet on egg
quality parameters, with the principal factor on quality
being the hen age. In the high-oleic peanut diet, there
was an impact on yolk color and β -carotene content
in eggs from the high-oleic peanut diet having darker
(P < 0.0001) yellow yolks in comparison to eggs from
the soybean meal diet (Table 4). After 1 wk of feeding
a high-oleic peanut diet, the yolk color DSM was signif-
icantly greater than the controls. This effect was more
pronounced over the experimental time period, with egg
yolk color score decreasing in intensity weekly of control
eggs, such that by week 9 the yolk color score of the con-
trol eggs were approximately 2-fold less than eggs from
the high-oleic peanut feeding group. In addition, there
was a significant temporal reduction in β -carotene con-
tent in eggs produced from hens fed the soybean meal
control diet from week 1 to 10 in comparison to eggs
produced from hens fed the high-oleic peanut diet over
time.
In summary, the predominant saturated fatty acids
found within experimental egg and feed samples were
palmitic fatty acid (16:0) and stearic fatty acid (18:0),
with concentrations of these saturated fatty acids
higher in feed samples and eggs produced from hens
fed the conventional soybean meal +corn diet. Ad-
ditionally, the unsaturated trans fatty acid that has
been implicated in heart disease (Tardy et al., 2011),
1746 TOOMER ET AL.

Figure 4. Immunoreactivity of protein extracts from pooled total egg samples from eggs produced by layer hens fed a diet with high-oleic
peanuts (HOPN) vs. a conventional diet with soybean meal (SBM) at week 10. Protein extracts from pooled egg samples (75 μ g per lane) were
run electrophoretically on a 10% polyacrylamide gel. To determine immunereactivity, resolved proteins were transferred to a nitrocellulose mem-
brane and immunoblotted with rabbit IgG anti-peanut agglutinin antibody (1:1000). Biodetection was determined with chromogenic peroxidase
substrate-based detection of HRP activity. (A) Immunoblotting results of total egg proteins from hen(s) #1 to 12 (HOPN) and hen(s) 13 to 24
(SBM). Extracted proteins (75 μ g) from peanut flour were utilized as a positive control. (B) Immunoblotting results of total egg proteins from
hen(s) #25 to 36 (SBM) and hen(s) 37 to 48 (HOPN). Extracted proteins (75 μ g) from peanut flour were utilized as a positive control.

n9 t elaidic, was significantly elevated in eggs produced
from layer hens fed the conventional soybean meal corn
diet. Overall, the predominant unsaturated fatty acids
found within the experimental egg and feed samples
were palmitoleic acid (omega 7 monounsaturated fatty
acid) and oleic acid (omega 9 monounsaturated fatty
acid), with contents of palmitoleic acid in higher con-
centrations in the feed and eggs produced from hens fed
the conventional soybean meal + corn diet. Conversely,
oleic acid content was greatest in the feed and egg sam-
ples from layer hens fed the high-oleic peanut + corn
diet.
Potential Transfer of Peanut Allergens in
Eggs produced in Layer Hens fed the
High-Oleic Peanut Diet
To determine the potential transfer of allergenic
peanut proteins to eggs produced from layer hens fed a
high-oleic peanuts + corn diet, protein extracts from
all eggs were analyzed by western immunoblotting
methods at weeks 1 (Figure 2), 5 (Figure 3), and 10
(Figure 4) experimentally. All egg protein extracts from
both treatments at each time point were non-reactive
with rabbit anti-peanut agglutinin antibodies, whereas
only protein samples from the positive control of peanut
flour were reactive. In summary, all egg samples were
non-allergenic for peanut antigens.
In contrast, earlier studies by Armentia et al. (2006)
and Faeste et al. (2014) reported hypersensitivity re-
actions in sensitized patients to allergens found in the
meat consumed of animals fed diets containing the aller-
genic proteins. Nevertheless, these studies investigated
the transfer of parasitic fish larva (Anisakis simplex,
parasitic nematode found in fish that causes allergic re-
action in some human consumers) allergenic peptides
from the diet to the meat produced, in contrast to ex-
amination of the transfer of proteins/peptides found
in feedstock rations. Therefore, this study aimed to
address the safety assessment of eggs produced from
layer hens fed a high-oleic peanut diet by utilizing
immunoblotting techniques with peanut-specific anti-
bodies. In summary, these studies demonstrate that
eggs produced from layer hens fed high-oleic peanuts
are non-reactive with peanut antibodies and there-
fore should not elicit an allergic response in peanut-
sensitized individuals. Furthermore, earlier research
has demonstrated that a great proportion of dietary
proteins and/or amino acids consumed in the diet
 EGGS FROM LAYER HENS FED HIGH-OLEIC PEANUTS
of livestock are either catabolized during first-pass
metabolism of the intestine, liver, intestinal cells, or
tissues (Stoll et al., 1998), and therefore are not found
directly in the eggs or meat produced.
In conclusion, this study helps validate the use of
high-oleic peanuts as a valuable feed ingredient for poul-
try and means to enrich the eggs produced with heart
healthy unsaturated fatty acids of health benefit to the
health conscious consumer.
SUPPLEMENTARY DATA
Supplementary data are available at Poultry Science
online.
ACKNOWLEDGMENTS
The authors would gratefully like to acknowledge
the following: Prestage Department of Poultry Science-
NCSU, NCSU Feed Mill, Birdsong Peanuts, Dr. Adam
Fahrenholz, and Market Quality & Handling Research
Unit for their contributions to this study and to Sab-
rina Whitley-Ferrell, Thien C. Vu and Rasi Fitria for
all of their administrative and technical assistance.
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