Resea ch A icle
Recei ed: 21 No embe 2019 Re ised: 4 Feb ua y 2020 Accep ed a icle published: 7 Feb ua y 2020 Published online in Wiley Online Lib a y: 24 Feb ua y 2020
(wileyonlinelib a y.com) DOI 10.1002/js a.10319
Supplemen a ion o lamb die s wi h i amin E
and osema y ex ac s on mea quali y
pa ame e s
Leonel N Leal,a,b
*
José A Bel án,cMa c Bellés,cJosé M Bello,d
Leo A den Ha og,a,b Wou e H Hend iksband Ja ie Ma ín-Te esoa
Abs ac
BACKGROUND: Sup anu i ional supplemen a ion o lamb die s wi h ⊍- ocophe ol is an e ec i e me hod o educe lipid oxida-
ion and colou de e io a ion in mea p oduc s. Howe e , al e na i e an ioxidan sou ces ha e been p oposed o eplace he
sup anu i ional i amin E applica ions.
RESULTS: Indoo concen a e- ed Rasa A agonesa male lambs (n=480) we e supplemen ed wi h inc easing le els o all- ac-
⊍- ocophe yl ace a e (0.25, 0.5, 1.0 g kg
−1
compound eed), osema y ex ac (0.20, 0.40, o 0.80 g kg
−1
compound eed), o
osema y ex ac embedded in a a ma ix (0.20, 0.40, o 0.80 g kg
−1
compound eed) o 14 days be o e slaugh e . The long-
issimus ho acis e lumbo um muscle om h ee lambs pe pen (18 lambs pe ea men ) we e modified-a mosphe e packaged
(70% O
2
+ 30% CO
2
) and main ained unde e ail condi ions o 14 days. Sup anu i ional supplemen a ion wi h an ioxidan s
had no e ec (P>0.05) on a e age daily weigh gain, eed in ake, and eed e ficiency. Rosema y ex ac supplemen a ion (wi h
o wi hou a embedmen ) had no e ec on lipid oxida ion, myoglobin o ms, o colou s abili y pa ame e s, ega dless o he
dose. All i amin E supplemen a ion le els significan ly a ec ed lipid oxida ion, colou s abili y (L*, C*, and h), myoglobin
o ms, and mea discolo a ion pa ame e s compa ed wi h non-supplemen ed lambs.
CONCLUSIONS: This s udy demons a es ha , unlike i amin E, nei he dose no p o ec ion o he osema y ex ac had an
e ec on lipid oxida ion o mea colou s abili y o lambs du ing he 14 days o s o age unde e ail condi ions.
© 2020 The Au ho s. Jou nal o The Science o Food and Ag icul u e published by John Wiley & Sons L d on behal o Socie y o
Chemical Indus y.
Keywo ds: i amin E; all- ac-⊍- ocophe yl ace a e; osema y ex ac ; lipid oxida ion; colou ; modified-a mosphe e packaging
INTRODUCTION
Appea ance, ex u e, and fla ou a e impo an quali y a ibu es
influencing he consume 's choice o pu chase mea p oduc s.
1
In lamb mea , pos -mo em biochemical changes, such as lipid
oxida ion, lead o o -odou s and fla ou de elopmen ha ha e
a nega i e impac on he shel li e o hese p oduc s.
2
The e o e,
he possibili y o ex end shel li e and subsequen display ime
in lamb mea is a p ima y objec i e o he mea indus y.
Die a y supplemen a ion o lamb die s wi h i amin E, and
mo e specifically ⊍- ocophe ol, aises he concen a ions o
⊍- ocophe ol in lamb issues,
3,4
which in u n delays lipid oxida ion
and imp o es he colou s abili y o he mea .
5–7
The p o ec i e ole
o ⊍- ocophe ol agains lipid oxida ion in lamb mea is well es ab-
lished. Howe e , he e is s ill a deba e on he minimum dose o i a-
min E in lamb die s ha e ec i ely p o ec s he mea om lipid
de e io a ion and alues anging om 287 and 1000 mg kg
−1
eed
ha e been p oposed.
5,7–10
Any ecommenda ion on i amin E
needs o conside he wide a ia ion o ⊍- ocophe ol concen a ion
in eeds u s,
11
i amin E s a us o he animal a he s a o he sup-
plemen a ion pe iod,
12
leng h o he supplemen a ion pe iod,
9
di e ences in slaugh e weigh ,
13
and animal b eed.
7
In Medi e a-
nean a ming sys ems, lambs a e ypically weaned a ound
12–14 kg o body weigh (BW), and a e wa ds ed ad libi um con-
cen a es plus s aw un il hey each 22–24 kg BW.
14
Unde hese
condi ions, Gonzalez-Cal o e al.
9
ound ha a concen a e supple-
men ed wi h 500 mg o i amin E pe kilog am, ed o a pe iod o
*Co espondence o: LN Leal, T ouw Nu i ion Resea ch and De elopmen , PO
Box 299, 3800 AG Ame s oo , Ne he lands. E-mail: leonel.
[email p o ec ed]
aT ouw Nu i ion Resea ch and De elopmen , Ame s oo , Ne he lands
bAnimal Nu i ion g oup, Depa men o Animal Sciences, Wageningen
Uni e si y & Resea ch, Wageningen, Ne he lands
cDepa amen o de P oducción Animal y Ciencia de los Alimen os, Facul ad de
Ve e ina ia, Ins i u o Ag oalimen a io de A agón (IA2), Za agoza, Spain
dNan a S.A., Mad id, Spain
© 2020 The Au ho s. Jou nal o The Science o Food and Ag icul u e published by John Wiley & Sons L d on behal o Socie y o Chemical Indus y.
This is an open access a icle unde he e ms o he C ea i e Commons A ibu ion License, which pe mi s use, dis ibu ion and ep oduc ion in any
medium, p o ided he o iginal wo k is p ope ly ci ed.
2922
7–14 days be o e slaugh e , was su ficien o p o ec lamb mea
om lipid oxida ion and me myoglobin (Me Mb) o ma ion.
Al e na i e an ioxidan sou ces o i amin E om se e al ypes
o plan s and plan ma e ials ha e been in es iga ed o eplace
i amin E. Howe e , o e ec i ely eplace sup anu i ional i amin
E applica ions, hese plan -de i ed an ioxidan s need o be e ec-
i ely abso bed, anspo ed, and ha e a conside able deposi ion
in o he lamb muscle. In ecen yea s, special a en ion has been
gi en o osema y (Rosma inus o ficinalis L.) p oduc s and by-
p oduc s, such as lea es, essen ial oils, dis illed lea es, and
ex ac s om he second dis illa ion o osema y lea es wi h di -
e en sol en s.
15–19
Ea ly wo k om Nie o e al.
16
ound ha sup-
plemen a ion o p egnan ewes wi h 100 g o osema y lea es
kilog am eed du ing p egnancy and lac a ion, imp o ed he shel
li e o lamb mea . In e es ingly, wo majo di e penes p esen in
osema y, ca nosic acid and ca nosol, we e la e iden ified in lamb
muscle a concen a ions ha could po en ially lead o an an iox-
idan and an imic obial e ec in he mea .
20
Supplemen a ion o
lamb die s wi h osema y by-p oduc s has yielded some conflic -
ing esul s,
19,21,22
p obably due o a lack o p oduc s anda diza-
ion
17
o because o po en ial umen deg ada ion.
23,24
The e a e only a ew s udies compa ing sup anu i ional doses
o i amin E wi h osema y by-p oduc s on hei capaci y o p o-
ec lamb mea om oxida i e de e io a ion.
17,25
Mo eo e , none
o he s udies pe o med a dose i a ion o bo h an ioxidan
sou ces in o de o de e mine he ela i e alue o each supple-
men a ion s a egy on mea quali y pa ame e s.
The p ima y aim o he p esen s udy is o compa e inc easing
dosages o a s anda dized osema y ex ac (RE) wi h i amin E
(all- ac-⊍- ocophe yl ace a e) in hei abili y o imp o e mea colou
and s abili y pa ame e s in lamb mea s o ed o 14 days unde
common e ail condi ions. Second, as a means o mi iga e he
deg ada ion o he RE by umen mic oo ganisms, a s anda dized
plan ex ac embedded in a umen-ine a will be also es ed.
MATERIALS AND METHODS
The animal ca e and slaugh e p ocedu es used me he guide-
lines o Council Di ec i e 86/609/EEC (Eu opean Communi ies,
1986) on he p o ec ion o animals used o expe imen al and
o he scien ific pu poses.
Animals and die s
A o al o 480 Rasa A agonesa male lambs we e used in his s udy
(BW =21.8 ±1.39 kg). E e y 3 weeks, lambs we e pu chased om
local deale s and inco po a ed in he expe imen in h ee sepa-
a ed ba ches o 160 animals each. Upon a i al a he comme cial
a m F anco y Na a o (Za agoza, Spain), lambs we e alloca ed
based on BW o 20 conc e e pens (20 m
2
) bedded wi h s aw.
Fo 14 days be o e slaugh e , he lambs had ee access o he
expe imen al compound eed (p esen ed as 3.5 mm diame e
pelle ; Table 1), ba ley s aw, and wa e ia sepa a ed oughs.
Pe ba ch, wo pens we e andomly assigned o one o 10 ea -
men s consis ing o a basal compound eed (con ol), supplemen-
ed wi h inc easing le els o all- ac-⊍- ocophe yl ace a e (0.25,
0.50, o 1.0 g kg
−1
compound eed), RE (Nu ox OSP; Nu a u
S.A., Mu cia, Spain; 0.20, 0.40, o 0.80 g kg
−1
compound eed), o
he same RE embedded in a a ma ix (FRE; 0.20, 0.40, o
0.80 g kg
−1
compound eed).
The di e en an ioxidan sou ces we e included in he com-
pound eed ia a ea men -specific expe imen al p emix
(Table 1). To p oduce he FRE, hyd ogena ed palm a y acids
(No el, Mad id, Spain) we e mel ed a 70 ±2°C. A e wa ds, he
RE (Nu ox OSP; Nu a u S.A.) was added o he mel ed
Table 1. Composi ion o he basal die and expe imen al p emix
Ing edien s (g kg
−1
)
Ba ley 295 Limes one 25
Soya bean meal (480 g kg
−1
o c ude p o ein) 240 Expe imen al p emix 10
Whea 200 Sodium bica bona e 7
Maize 200 Sodium chlo ide 5
Soya oil 15 S anda d mine al and i amin p emix
a
3
SE RE FRE
Expe imen al p emix Con ol 250 500 1000 200 400 800 200 400 800
Ing edien s, g kg
−1
Whea b an 560 560 560 560 560 560 560 560 560 560
Hyd opalm
b
140 140 140 140 140 140 140 105 70
Silica
c
300 250 200 100 280 260 220 255 210 120
Rosema y
d
20 40 80
Fa -embedded osema y
e
80 160 320
all- ac-⊍-Tocophe yl ace a e
50 100 200
a
Mine al and i amins p o ided: calcium 0.24 g, sodium 0.47 g, sul u 0.23 g, manganese 30 mg, zinc 50 mg, coppe 5 mg, iodine 0.5 mg, cobal
0.5 mg, selenium 0.15 mg, i on 50 mg, i amin A 8000 IU, i amin D
3
1600 IU, all- ac-⊍- ocophe yl ace a e 25 mg. SE: all- ac-⊍- ocophe yl ace a e.
RE: osema y ex ac . FRE: a -embedded osema y ex ac .
b
Hyd opalm: hyd ogena ed palm a y acids (No el, Spain).
c
Silica: silica H
2
O (T ouw Nu i ion, Ne he lands).
d
Rosema y: s anda dized osema y ex ac (Nu a u S.A., Spain).
e
Fa -embedded osema y: s anda dized osema y ex ac (inclusion o 25%) mixed wi h mel ing (70 ±2°C) hyd opalm (43.75%) and silica (31.25%).
all- ac-⊍-Tocophe yl ace a e: con ains 50 g all- ac-⊍- ocophe ol pe 100 g o p oduc (T ouw Nu i ion, Ne he lands).
Die a y i amin E and osema y ex ac s on mea quali y in lambs www.soci.o g
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Jou nal o The Science o Food and Ag icul u e published by John Wiley & Sons L d on behal o Socie y o Chemical Indus y.
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2923
hyd ogena ed palm a y acids and s i ed o 10 min a 70 ±2°C,
un il an homogeneous mix u e was ob ained. The mix u e o
hyd ogena ed palm a y acids and RE was hen added o silica
(T ouw Nu i ion, Ame s oo , Ne he lands), s i ed o 20 min
and cooled a oom empe a u e. The final p oduc ( ee-flowing
powde ) was composed o 43.75% hyd ogena ed palm a y acids,
31.25% silica, and 25.0% RE.
A e 14 days o exposu e o he ea men die s and ollowing
an o e nigh pe iod wi hou eed (bu wi h ee access o wa e ),
lambs om all ea men s we e mixed in a single g oup, ans-
po ed in he same lo y, and slaugh e ed a he local aba oi
(Me caza agoza S.A., Za agoza, Spain) wi hin 2 h a e lea ing
he a m.
Mea p ocessing and packaging
Th ee lambs pe pen (in o al, 18 lambs pe ea men ) we e an-
domly selec ed a he s a o he expe imen al pe iod o mea
analysis. The ca casses we e chilled o 24 h a 2 °C be o e being
spli longi udinally in o wo hal es. The longissimus ho acis e
lumbo um (LTL) muscle om he igh hal was dissec ed, wi h
subcu aneous a emo ed, placed in bags (one muscle pe
bag), and anspo ed in sealed plas ic con aine s in da kness a
4±1°C o he Mea Quali y Labo a o y o he Ve e ina y Facul y
o Za agoza (Za agoza, Spain).
Wi hin 2 h a e a i al, he LTL muscles we e sec ioned in o
app oxima ely 1.5 cm hick po ions, which we e placed in
polys y ene ays B5-37 (Ae pack), packed unde modified-
a mosphe e packaging wi h 70% oxygen (O
2
) + 30% ca bon diox-
ide (CO
2
) (Ulma Sma 500; Ulma Packaging, Guipúzcoa, Spain),
and sealed wi h a polye hylene and polyamine lamina e film
(30 μm hickness, wa e apou ansmission a e a 23 °Co
<7gm
−2
pe 24 h a 85% ela i e humidi y (RH), an O
2
ansmis-
sion a e a 23 °Co <15 cm
3
m
−2
a 0% RH, and a CO
2
ansmis-
sion a e a 23 °Co <75 cm
3
m
−2
pe 24 h a 0% RH; Linpac
Packaging S.L., Spain). The ays we e s o ed a 4 ±1°C o 1, 7,
9, 12, o 14 days in a Za io display cabine (Za io, Za agoza,
Spain), simula ing e ail condi ions wi h a daily exposu e o 14 h
o ligh a 1200 lx.
Physical and chemical analysis
Colou was measu ed using a eflec ance spec opho ome e
(CM-2002; Minol a, Osaka, Japan) di ec ly on he mea su ace,
a e 2 h exposu e o ai . Each alue was he mean o 10 de e mi-
na ions pe sample. The pa ame e s eco ded acco ding o he
CIELAB sys em,
26
we e ligh ness L*, edness a*, and yellowness
b*. Values o ch oma C* and hue angle hwe e calcula ed as
C*=(a*
2
+b*
2
)
0.5
, and h= an
−1
(b*/a*), exp essed in deg ees.
Lipid oxida ion was exp essed as hioba bi u ic acid eac i e
subs ances (TBARS) and exp essed as millig ams o malondialde-
hyde (MDA) pe kilog am o mea , as desc ibed by P alzg a
e al.
27
B iefly, 10 g o mea sample was homogenized wi h 10%
ichlo oace ic acid using an Ul a-Tu ax T25 (Janke & Kunkel,
S au en, Ge many). A e cen i uga ion a 1500×g o 30 min
(a 10 °C) he supe na an was fil e ed (Fil e lab, Ba celona, Spain),
2 mL o he fil a e was mixed wi h 2 mL o hioba bi u ic acid
(20 mol L
−1
), homogenized, and incuba ed o 20 min (in boiling
wa e ). Abso bance was measu ed a 532 nm, and TBARS alues
we e calcula ed om a s anda d cu e o MDA ob ained by he
hyd olysis o 1,1,3,3- e ame hoxyp opane (Sigma-Ald ich,
Mad id, Spain). Samples we e analysed in duplica e, and he
esul s a e exp essed as millig ams o MDA pe kilog am o mea .
Myoglobin o ms (deoxymyoglobin, oxymyoglobin (OxyMb),
and Me Mb) we e calcula ed om he eflec ance cu e desc ibed
by K zywicki
28
using a wa eleng h o 690 nm. The eflec ance a
473, 525, and 572 nm was ob ained by linea in e pola ion, since
he eflec ance spec opho ome e only measu es he eflec ance
be ween 400 and 740 nm a in e als o 10 nm.
The a e o mea discolo a ion was accessed by he pa ame e
A
580
−A
630
, acco ding o he me hod p oposed by an den Oo d
and Wesdo p
29
Oxygen sa u a ion o myoglobin on mea su ace
I
SO2
was measu ed ollowing he echnique desc ibed by Tsu uga
e al.
30
Polyphenol analysis in eed
Feed samples we e homogenized using a blende and sie ed wi h
a no. 18 mesh (co esponding o 1 mm) o emo e la ge pa icles
be o e ex ac ion. The phenolic compounds we e ex ac ed using
me hanol:wa e (80:20, : ), sonica ed o 5 min, and cen i uged.
This p ocedu e was epea ed wice wi h he eed esidue. The
supe na an s we e combined and he me hanol e apo a ed. Fo
be e ch oma og aphy esolu ion, he ex ac s we e cleaned
using solid-phase ex ac ion wi h Oasis MAX 96-well pla es
(30 mg, 30 μm) om Wa e s (Wa e co po a ion, Massachuse s,
USA), acco ding o Vall e dú-Que al e al.
31
The iden ifica ion and quan ifica ion o he majo phenolic
compounds was pe o med using ul ahigh pe o mance liquid
ch oma og aphy (UHPLC; Wa e s Acqui y Ul a Pe o mance LC)
coupled o an API 3000 iple quad upole mass spec ome e
(ABSciex) equipped wi h a Tu bo ion sp ay sou ce ope a ing in
nega i e mode. Sepa a ion was ca ied ou using a BEH C18
(2.1 mm ×50 mm, 1.7 μm; Acqui y UHPLC) main ained a 30 °
C. G adien elu ion was pe o med wi h ace oni ile wi h 0.1%
o o mic acid ( / ) and wa e wi h 0.1% o o mic acid ( / )
using an inc easing linea g adien flow o ace oni ile wi h
0.1% o o mic acid ( / ) unde he ollowing condi ions: 0 min,
20%;0.5min,20%;1.5min,30%;2min,30%;2.5min,50%;
3min,100%;3.5min,100%;3.7min,20%;and4.5min,20%.
The flow a e was 0.4 mL min
−1
and he injec ion olume was
10 μL. The iden ifica ion and quan ifica ion o each compound
we e ca ied ou using a mass spec ome e in mul iple eac ion
moni o ing mode. The quan ifica ion o he phenolic com-
pounds was pe o med using calib a ion cu es wi h analy ical
s anda ds wi h he in e nal s anda d me hod. The in e nal s an-
da d was e hyl galla e (400 ng g
−1
) (Ex asyn hese, Genay,
F ance) and he esul s a e exp essed as millig ams pe kilog am
o eed.
32
S a is ical analysis
All s a is ical analyses we e pe o med using SAS S udio (SAS Ins i-
u e, Inc., Ca y, NC, USA). Indi idual lamb da a we e summa ized
pe pen, which was conside ed as he expe imen al uni o all
o he pa ame e s s udied.
The model included he fixed e ec s o an ioxidan supple-
men a ion, display ime, and he in e ac ion be ween an ioxi-
dan supplemen a ion and display ime. The e ec o ba ch
( h ee ounds o 180 lambs each) was ini ially included in he s a-
is ical model as a fixede ec ,bu i wasfinally excluded as
ba ch ne e eached s a is ical significance (P>0.05). The e o e,
he model was
Yijk =⊘+ASi+Dj+AS
i×Dj
+eijk
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J Sci Food Ag ic 2020; 100: 2922–2931
2924
whe e Y
ijk
is he dependen a iable, ⊘is he popula ion a e age,
AS
i
is he fixed e ec o an ioxidan supplemen a ion, D
j
is he
fixed e ec o display ime, AS
i
×D
j
is he in e ac ion be ween
AS
i
and D
j
, and e
ijk
is he andom e o . Di e ences we e decla ed
significan when P<0.05. Tukey's pos hoc es was used o
assess di e ences be ween mean alues when P<0.05.
RESULTS AND DISCUSSION
Animal pe o mance
Da a on BW, a e age daily weigh gain, eed in ake, and eed e fi-
ciency a e p esen ed in Table 2. The e we e no e ec s o ei he
an ioxidan sou ce ( i amin E and RE) o dosage in he assessed
pe o mance pa ame e s. These esul s align wi h p e ious find-
ings in which die a y i amin E inclusion le els up o 2.0 g kg
−1
eed had no e ec on lamb pe o mance.
3,5
Mo eo e , supple-
men a ion o lamb die s wi h osema y di e penes
17
(0.6 g kg
−1
die ) o osema y essen ial oils
19
(0.3 o 0.6 mL day
−1
) we e also
ound o ha e no e ec on g ow h, eed in ake, o e ficiency.
Polyphenol con en in eeds and a embedmen
The iden ifica ion and quan ifica ion o he majo polyphenols
p esen in he expe imen al die s a e shown in Table 3. O e all,
we ound high concen a ions o chlo ogenic acid, e ulic acid,
and e ulic acid-O-hexoside, which a e commonly ound in
ce eals,
33
which accoun ed o >42% o he o al polyphenol
con en in he expe imen al die s. Die s con aining REs (ei he
p o ec ed o unp o ec ed), p o ided mo e ca nosol, ca nosic,
and osma inic acid, majo polyphenols p esen in REs,
34
han
he con ol die . Acco ding o he in o ma ion p o ided by
he supplie , he RE con ained app oxima ely 20% di e penes,
in a a io o 2:1 be ween ca nosic acid and ca nosol. Howe e ,
RE supplemen a ion (RE and FRE) esul ed in highe concen a-
ions o ca nosol han ca nosic acid in he expe imen al eeds.
Acco ding o Jo dán e al.,
20
ca nosol is a majo componen
p oduced in he ca nosic acid oxida ion pa hway, which could
explain he high ca nosol concen a ions in he osema y-
supplemen ed die s and he ela i e low amoun s o ca nosic
acid. On he o he hand, he lowe amoun s o ca nosol and
Table 2. P oduc i e pe o mance o he lambs ed inc easing le el o all- ac-⊍- ocophe yl ace a e (SE), osema y ex ac (RE), o a -embedded ose-
ma y ex ac (FRE)
SE (mg kg
−1
eed) RE (mg kg
−1
eed) FRE (mg kg
−1
eed)
SEM P alueI em Con ol 250 500 1000 200 400 800 200 400 800
IBW (kg) 21.9 21.8 21.8 21.9 21.9 21.9 21.8 21.8 21.8 21.9 0.04 0.746
FBW (kg) 26.4 26.2 26.5 26.4 26.2 26.7 26.5 26.4 26.3 26.5 0.29 0.848
FEED, (kg day
−1
) 0.992 0.996 1.005 1.015 1.006 1.039 1.032 1.025 1.029 1.027 0.0362 0.880
ADG (kg day
−1
) 0.329 0.315 0.331 0.324 0.315 0.351 0.331 0.323 0.319 0.331 0.0205 0.832
FCR (kg kg
−1
) 3.11 3.25 3.07 3.17 3.20 3.07 3.21 3.15 3.40 3.21 0.191 0.824
ADG: a e age daily gain; FBW: final body weigh ; FCR: eed con e sion a io; FEED: compound eed in ake; IBW: ini ial body weigh ; SEM: s anda d
e o o he mean.
Table 3. Quan ifica ion o indi idual bioac i e compounds (mg kg
−1
) o he di e en eeds
SE (mg kg
−1
eed) RE (mg kg
−1
eed) FRE (mg kg
−1
eed)
I em Con ol 250 500 1000 200 400 800 200 400 800
CA 261 357 223 255 264 314 266 235 185 265
C A 13 13 13 13 23 19 24 25 19 27
Cn 11 8 6 10 1366 1383 2149 862 553 551
ChA 1060 1084 801 837 782 634 755 777 498 742
DA 55 7 4319181522111228
FA 926 845 810 716 753 675 872 785 594 851
FAH 1593 1423 1401 1264 1375 858 1677 1649 1035 1761
N 60 676768626463695560
NG 373 392 367 355 362 338 379 379 292 363
pCA 596 491 489 429 436 444 522 480 356 528
PA 166 228 140 154 156 166 171 159 107 158
Q 55 585756575862595856
RsA 31 29 30 31 40 71 128 35 50 40
R 118 73 101 73 66 92 79 76 79 83
To al 5318 5075 4548 4280 5760 5131 7169 5601 3893 5513
CA: ca eic acid; ChA: chlo ogenic acid; Cn: ca nosol; C A: ca nosic acid; DA: dica eoylquinic acid; FA: e ulic acid; FAH: e ulic acid-O-hexoside; FRE: a -
embedded osema y ex ac ; N: na igenin; NG: na ingenin glucoside; PA: p o oca echuic acid; pCA: p-cuma ic acid; Q: que ce in; R: u in; RE: osema y
ex ac ; RsA: osma inic acid; SE: all- ac-⊍- ocophe yl ace a e.
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ca nosic acid in FRE die s compa ed wi h RE die s could be
associa ed wi h he ex a hea ea men
35
applied o he FRE
die s du ing he inco po a ion o he RE in o he mel ed a
(70 ±2°C).
Lipid oxida ion
Mean TBARS alues o each die a y ea men du ing he 14 days
o display a e p esen ed in Table 4. Lipid oxida ion significan ly
inc eased wi h s o age ime in all g oups (P<0.05). Mo eo e ,
an in e ac ion be ween s o age ime and die a y ea men was
ound (P<0.001) o TBARS alues. F om day 7 onwa ds, LTL
muscle om i amin-E-supplemen ed lambs (i espec i e o dose)
p esen ed significan ly lowe TBARS alues han he con ol and
RE‑and FRE-supplemen ed lambs. Vi amin-E-supplemen ed
lambs p esen ed TBARS alues anging om 0.16 o 0.31 mg
MDA kg
−1
mea on day 7, whe eas he emaining ea men s eg-
is e ed TBARS alues be ween 1.63 and 1.99 mg MDA kg
−1
mea .
Wi h inc easing s o age ime, he di e ences be ween he
i amin-E-supplemen ed lambs and he con ol, RE, and FRE
lambs became wide . On day 14 o s o age, LTL muscle om con-
ol and he osema y-supplemen ed lambs (RE and FRE) p e-
sen ed TBARS alues anging om 3.25 o 4.00 mg MDA kg
−1
mea , whe eas i amin-E-supplemen ed LTL muscles showed
TBARS alues om 0.39 o 0.69 mg MDA kg
−1
mea . In
concen a e- ed lambs, slaugh e ed a low BWs (lowe han
30 kg li e weigh ), a h eshold o 1.0 mg MDA kg
−1
mea has been
p oposed o be a oided o he de elopmen o o fla ou s in
lamb mea .
36
A e 7 days o s o age, highe MDA alues han
1.0 mg kg
−1
mea we e ound in he LTL muscle om he con ol,
FRE, and RE lambs. In e es ingly, i amin E supplemen a ion a
0.25, 0.50 o 1.0 g kg
−1
eed led o lowe TBARS alues (0.69,
0.56, and 0.39 mg MDA kg
−1
mea ) han he p oposed h eshold
a e 14 days o s o age. In a simila s udy, Leal e al.
6
ound ha
a supplemen a ion le el o 0.16 g kg
−1
eed wi h i amin E (all-
ac-⊍- ocophe yl ace a e) o 14 days be o e slaugh e was su fi-
cien o main ain TBARS alues below he h eshold o 1.0 mg
MDA kg
−1
mea . Die a y supplemen a ion o lamb die s wi h
i amin E is a consis en s a egy o inc ease ⊍- ocophe ol con en
in lamb issues like muscle,
3
and o imp o e he esis ance o lamb
mea o oxida i e de e io a ion.
5,6,37
Unlike i amin E, he e ec s o REs o essen ial oils on mea
oxida i e s abili y a e no consis en . Shel -li e s udies in lambs
ha e demons a ed an imp o emen in lipid s abili y o mea
when osema y by-p oduc s we e included in he die o lac a -
ing ewes
16
o di ec ly in o lamb die s.
15,38
In he cu en s udy,
supplemen a ion o lamb die s wi h REs (RE and FRE) ailed o
a ec he oxida i e de e io a ion o mea . Acco dingly, Sme i
e al.
18
and Aouadi e al.
21
ound ha supplemen ing lambs
wi h osema y essen ial oils did no exe any e ec on lipid
pe oxida ion in mea . O e all, he mechanisms by which an i-
oxidan compounds p esen in plan ex ac s can a ec he
oxida i e de e io a ion o lamb mea a e s ill unclea .
39
In e -
es ingly, se e al s udies ound a highe concen a ion o
ocophe ol in lamb muscle ollowing die a y supplemen a ion
wi h a plan ex ac s o by-p oduc .
17,40,41
This is o special el-
e ance, because e en a sligh inc ease in ⊍- ocophe ol concen-
a ion in muscle ( om 0.6 o 0.9 mg kg
−1
mea ) has been
shown o subs an ially dec ease lipid oxida ion a 5 and 7 days
o display.
9
Colou s abili y
Resul s o he colou measu emen s a e shown in Table 5. O e all,
s o age ime significan ly a ec ed he e olu ion o all mea colou
pa ame e s. Mo eo e , in e ac ions be ween he die a y ea -
men and s o age ime we e egis e ed o L* (ligh ness), C*
(ch oma), and h(hue angle) alues (P<0.001).
A e 12 days o s o age, a clea educ ion in L* alues was ound
in he mea o i amin-E-supplemen ed lambs ( ega dless o
dose). When compa ed wi h he con ol, LTL muscle om lambs
supplemen ed wi h RE o FRE p esen ed no significan imp o e-
men s in L* alues. I has been p e iously p oposed
42
ha human
e alua o s a e no able o app ecia e indi idual L*,a*, and b* coo -
dina es. The e o e, hue angle hp o ides a be e es ima ion o
mea b owning han indi idual coo dina es. Like p e iously
Table 4. E ec o die a y supplemen a ion
a
wi h all- ac-⊍- ocophe yl ace a e (SE), osema y ex ac (RE), and a -embedded osema y ex ac (FRE)
on hioba bi u ic acid eac i e subs ances in aw lamb mea s o ed in modified-a mosphe e packaging (70% O
2
:30% CO
2
) kep o 1, 7, 9, 12, and
14 days unde e ail condi ions
I em
Days o display
SEM P- alue display1791214
Con ol 0.13
c,x
1.90
b,y
2.66
b,yz
3.44
b,z
3.80
b,z
0.25 <0.001
SE 250 0.09
ab,x
0.31
a,xy
0.32
a,xy
0.41
a,y
0.69
a,z
0.06 <0.001
SE 500 0.08
a,x
0.21
a,y
0.26
a,y
0.30
a,y
0.56
a,z
0.03 0.001
SE 1000 0.09
ab,x
0.16
a,xy
0.18
a,xy
0.33
a,yz
0.39
a,z
0.04 0.017
RE 200 0.11
abc,w
1.99
b,x
2.85
b,xy
3.53
b,yz
4.00
b,z
0.23 <0.001
RE 400 0.11
abc,w
1.91
b,x
2.75
b,xy
3.61
b,yz
3.85
b,z
0.21 <0.001
RE 800 0.11
abc,w
1.86
b,x
2.55
b,xy
3.64
b,yz
3.80
b,z
0.24 <0.001
FRE 200 0.11
abc,x
1.67
b,y
2.46
b,yz
2.80
b,yz
3.25
b,z
0.30 <0.001
FRE 400 0.11
abc,w
1.63
b,x
2.29
b,xy
3.27
b,yz
3.51
b,z
0.23 <0.001
FRE 800 0.11
abc,x
1.74
b,y
2.50
b,yz
3.25
b,z
3.53
b,z
0.23 <0.001
P- alue ea men 0.016 <0.001 <0.001 <0.001 <0.001
a
SE 250, SE 500, and SE 1000: 250, 500, and 1000 mg all- ac-⊍- ocophe yl ace a e kg
−1
eed; RE 200, RE 400, and RE 800: 200, 400, and 800 mg ose-
ma y ex ac kg
−1
eed; FRE 200, FRE 400, and FRE 800: 200, 400, and 800 mg a -embedded osema y ex ac kg
−1
eed.
a,b,c
Values wi hin a column wi h di e en supe sc ip a e significan ly di e en (P<0.05).
w,x,y,z
Values wi hin a ow wi h di e en supe sc ip a e sig-
nifican ly di e en (P<0.05). SEM: s anda d e o o he mean.
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2926
epo ed o indi idual L* coo dina es, i amin E supplemen a ion
significan ly a ec ed hde elopmen in lamb mea when com-
pa ed wi h he con ol‑,RE‑, and FRE- ed lambs. The e ec s on
h alue a e clea a e 9 days o display. No e ec was ound o
RE supplemen a ion in h alues compa ed wi h he con ol.
Ch oma C* in mea , like wi h he L* alues, was only a ec ed by
he die a y supplemen a ions a e 12 days o s o age. Indepen-
den ly om dosage, i amin E supplemen a ion led o a signifi-
can inc ease in C* alues compa ed wi h he mea o con ol‑,
RE‑, and FRE- ed lambs.
O e all, he posi i e e ec o syn he ic
6,43
and na u al
6
i amin E
supplemen a ion on mea colou pa ame e s has al eady been
desc ibed in lambs. Supplemen a ion o lamb die s wi h i amin
E le els abo e 0.5 g kg
−1
eed has been associa ed wi h signifi-
can imp o emen s in L*,
39
C,*
17
and h alues
6
in mea , when
compa ed wi h non-supplemen ed lambs. Howe e , ou findings
conflic wi h ea lie s udies ha epo ed a posi i e e ec on
L*,
17,38
C*,
38
and h alues
38,44
in lamb mea , ollowing a die a y
supplemen a ion wi h REs o hei di e penes. Like wi h
i amin E, o he phenolic compounds o exhibi any di ec an i-
oxidan ac i i y pos -mo em hey need o be abso bed along he
gas oin es inal ac , anspo ed in he blood, and accumula ed
in he a ge issue (such as he muscle).
45
The e o e, ac o s ha
can a ec he concen a ion o phenolic compounds in muscle,
Table 5. E ec o die a y supplemen a ion
a
wi h all- ac-⊍- ocophe yl ace a e (SE), osema y ex ac (RE), and a -embedded osema y ex ac (FRE)
on colou pa ame e s (L*,C*, h) in aw lamb mea s o ed in modified-a mosphe e packaging (70% O
2
:30% CO
2
) kep o 1, 7, 9, 12, and 14 days unde
e ail condi ions
Days o display
SEM P- alue displayI em 1 7 9 12 14
Ligh ness L*
Con ol 40.94
x
39.19
x
40.17
ab,x
45.26
b,y
47.67
b,z
0.269 <0.001
SE 250 40.22
yz
36.74
w
37.98
ab,wx
38.78
a,xy
40.78
a,z
0.173 <0.001
SE 500 40.32
y
37.85
x
38.04
ab,x
39.21
a,xy
40.55
a,y
0.256 <0.001
SE 1000 39.52
yz
37.31
x
37.68
a,xy
38.71
a,xyz
40.67
a,z
0.221 <0.001
RE 200 40.11
x
38.45
x
40.00
ab,x
44.82
b,y
47.33
b,y
0.359 <0.001
RE 400 39.41
x
38.38
x
40.24
ab,x
45.12
b,y
48.73
b,z
0.406 <0.001
RE 800 39.17
wx
38.15
w
40.67
b,x
44.92
b,y
48.42
b,z
0.252 <0.001
FRE 200 39.95
x
38.39
x
40.09
ab,x
43.39
b,y
46.49
b,z
0.324 <0.001
FRE 400 40.85
x
38.58
x
39.93
ab,x
44.19
b,y
47.09
b,z
0.389 <0.001
FRE 800 40.98
x
37.85
w
40.66
b,wx
44.98
b,y
48.71
b,z
0.356 <0.001
P- alue ea men 0.530 0.146 0.001 <0.001 <0.001
Ch oma C*
Con ol 13.39
x
15.49
ab,y
13.54
a,x
13.15
a,x
13.55
a,x
0.171 <0.001
SE 250 14.84
x
16.92
b,yz
17.12
d,z
16.18
c,yz
15.94
b,y
0.122 <0.001
SE 500 13.94 16.10
ab
15.65
bcd
14.95
b
15.38
b
0.156 0.059
SE 1000 13.85
x
16.12
ab,y
16.03
cd,y
15.73
bc,y
16.08
b,y
0.131 0.002
RE 200 13.93 14.81
a
13.37
a
12.95
a
13.77
a
0.179 0.189
RE 400 13.97
xy
14.66
a,y
13.75
a,xy
12.98
a,x
13.74
a,xy
0.171 0.040
RE 800 13.20
x
15.42
a,y
13.99
ab,x
12.87
a,x
13.55
a,x
0.153 <0.001
FRE 200 14.84 14.89
ab
14.29
abc
13.60
a
13.96
a
0.162 0.245
FRE 400 13.62
xy
15.14
a,z
14.32
abc,yz
12.55
a,x
13.58
a,xy
0.154 0.006
FRE 800 14.17
xy
15.36
a,y
13.86
ab,x
12.91
a,x
13.56
a,x
0.156 <0.001
P- alue ea men 0.077 0.003 <0.001 <0.001 <0.001
Hue angle h
Con ol 51.41
x
50.64
abc,x
58.85
b,x
74.04
b,y
77.15
b,y
1.120 <0.001
SE 250 50.15
z
44.36
a,x
46.16
a,xy
46.69
a,y
48.96
a,z
0.238 <0.001
SE 500 51.15
z
45.78
abc,x
45.72
a,x
47.37
a,xy
48.71
a,yz
0.339 <0.001
SE 1000 50.68
z
44.60
ab,x
46.25
a,xy
47.10
a,xy
48.17
a,yz
0.288 <0.001
RE 200 50.24
x
52.05
c,xy
61.13
b,y
73.90
b,z
76.95
b,z
1.185 <0.001
RE 400 50.38
x
51.54
bc,x
61.73
b,y
76.42
b,z
79.26
b,z
1.145 <0.001
RE 800 50.74
x
48.40
abc,x
60.00
b,y
73.98
b,z
79.29
b,z
0.936 <0.001
FRE 200 50.52
x
50.74
abc,x
57.74
b,x
68.23
b,y
73.35
b,y
1.151 <0.001
FRE 400 52.30
x
50.35
abc,x
55.32
ab,x
70.54
b,y
76.03
b,y
1.156 <0.001
FRE 800 51.08
xy
49.94
abc,x
58.96
b,y
73.93
b,z
78.78
b,z
1.040 <0.001
P- alue ea men 0.883 <0.001 <0.001 <0.001 <0.001
a
SE 250, SE 500, and SE 1000: 250, 500, and 1000 mg all- ac-⊍- ocophe yl ace a e kg
−1
eed; RE 200, RE 400, and RE 800: 200, 400, and 800 mg ose-
ma y ex ac kg
−1
eed; FRE 200, FRE 400, and FRE 800: 200, 400, and 800 mg a embedded osema y ex ac kg
−1
eed.
a,b,c
Values wi hin a column wi h di e en supe sc ip a e significan ly di e en (P<0.05).
w,x,y,z
Values wi hin a ow wi h di e en supe sc ip a e sig-
nifican ly di e en (P<0.05). SEM: s anda d e o o he mean.
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such as dosage and leng h o he supplemen a ion pe iod, may
explain he esul s in mea colou and oxida i e s abili y o he RE‑
and FRE-supplemen ed lambs.In he cu en s udy, lambs we e sup-
plemen ed o 14 days wi h a die con aining inc easing le els (0.2,
0.4 o 0.8 g kg
−1
) o an RE (RE o FRE) ha con ained app oxima ely
20% di e penes (acco ding o manu ac u e 's specifica ions). In con-
as , s udies ha epo ed posi i e e ec s o supplemen ing lamb
die s wi h REs (o di e penes) on mea colou s abili y a ge ed lon-
ge supplemen a ion pe iods,
15,16
highe dosages,
44,46
and/o we e
pe o med using ex ac s wi h highe pu i y.
17
Myoglobin oxida ion and mea discolo a ion
The esul s o mea myoglobin (Me Mb and OxyMb) oxida ion a e
p esen ed in Table 6. O e all, myoglobin oxida ion ou comes a e
consis en wi h he colo ime ic pa ame e s p e iously discussed.
Display ime significan ly a ec ed all he myoglobin o ms ana-
lysed (P<0.01). Mo eo e , an in e ac ion be ween an ioxidan
supplemen a ion and s o age ime was ound o Me Mb and
OxyMb (P<0.001). Supplemen a ion o lamb die s wi h i amin
E (0.25, 0.50 o 1.0 g kg
−1
eed) was ound o consis en ly a ec
Me Mb and OxyMb oxida ion a e 12 days o s o age. Howe e ,
RE p esen a ion o m (RE o FRE) o dosage (0.20, 0.40 o
0.8 g kg
−1
eed) had no e ec on Me Mb and OxyMb pe cen ages
in he LTL muscle when compa ed wi h he con ol. The e ec s o
i amin E supplemen a ion on Me Mb de elopmen a e consis-
en wi h p e ious wo k in lambs,
5,6
whe e a clea inhibi o y e ec
o i amin E supplemen a ion on Me Mb o ma ion in lamb mea
was epo ed, especially du ing longe display pe iods. Mo eo e ,
in line wi h he cu en s udy, Yagoubi e al.
41
ound no e ec o
supplemen ing lamb die s wi h osema y lea esidues on Me Mb
pe cen ages in mea displayed o 9 days. The p opo ion o
OxyMb inc eased significan ly in all g oups (P<0.01) un il 7 days
o s o age and dec eased he ea e un il day 14 o s o age. Apa
om display ime, supplemen a ion o lamb die s wi h i amin E
(0.25, 0.50 o 1.0 g kg
−1
eed) significan ly educed OxyMb oxida-
ion when compa ed wi h he o he ea men s a e 12 days o
display. These findings con as wi h p e ious wo k ha ound
no e ec o i amin E supplemen a ion on OxyMb oxida ion in
lambs.
5,6
In e es ingly, supplemen a ion o lamb die s wi h REs
(RE and FRE) had no e ec on OxyMb pe cen ages when com-
pa ed wi h he con ol g oup. Howe e , Yagoubi e al.
41
ound a
clea educ ion in OxyMb oxida ion in he LTL muscle o Ba ba ine
lambs supplemen ed wi h osema y dis illa ion esidues o
77 days be o e slaugh e .
Mea discolo a ion, as assessed by he dec ease in he
A
580
−A
630
pa ame e , and O
2
sa u a ion on he mea su ace
Table 6. E ec o die a y supplemen a ion
a
wi h all- ac-⊍- ocophe yl ace a e (SE), osema y ex ac (RE), and a -embedded osema y ex ac (FRE)
on myoglobin o ms (OxyMb and Me Mb) in aw lamb mea s o ed in modified-a mosphe e packaging (70% O
2
:30% CO
2
) kep o 1, 7, 9, 12, and
14 days unde e ail condi ions
Days o display
SEM P- alue displayI em 1 7 9 12 14
OxyMb
Con ol 28.24
y
41.68
z
37.04
yz
16.41
a,x
11.85
a,x
2.90 <0.001
SE 250 27.02
x
42.07
z
38.97
yz
38.10
bc,y
37.03
b,y
0.86 <0.001
SE 500 22.34
x
42.98
z
40.59
yz
38.60
c,yz
36.14
b,y
1.15 <0.001
SE 1000 25.53
x
41.57
z
38.97
yz
37.90
bc,y
37.29
b,y
0.91 <0.001
RE 200 26.39
xy
40.16
z
33.22
yz
20.03
a,wx
12.43
a,w
2.73 <0.001
RE 400 26.36
y
40.98
z
31.47
y
15.39
a,x
11.05
a,x
2.18 <0.001
RE 800 23.14
z
41.73
z
34.46
y
20.00
a,xy
12.39
a,x
2.32 <0.001
FRE 200 28.83
xy
39.67
z
36.16
yz
20.53
a,wx
16.23
a,w
2.40 <0.001
FRE 400 28.01
xy
39.86
z
33.88
yz
22.63
a,wx
17.04
a,w
3.00 0.005
FRE 800 24.52
x
40.67
z
34.99
yz
25.93
a,wx
13.72
a,w
2.30 <0.001
P- alue ea men 0.928 0.738 0.593 <0.001 <0.001
Me Mb
Con ol 22.73
w
33.95
x
46.55
b,y
61.06
b,z
63.56
b,z
2.50 <0.001
SE 250 22.77
w
29.23
x
30.72
a,xy
32.97
a,y
35.98
a,z
0.69 <0.001
SE 500 24.80
w
28.11
wx
30.55
a,xy
32.74
a,yz
36.10
a,z
0.82 <0.001
SE 1000 23.67
w
28.96
x
30.55
a,xy
32.38
a,y
34.35
a,z
0.62 <0.001
RE 200 23.37
x
36.62
y
48.54
ab,y
60.96
b,z
64.08
b,z
2.78 <0.001
RE 400 24.07
w
35.93
x
49.51
ab,y
64.61
b,z
65.93
b,z
2.26 <0.001
RE 800 21.58
w
31.87
x
47.19
b,y
60.86
b,z
65.17
b,z
2.01 <0.001
FRE 200 22.87
x
36.09
x
43.34
b,y
55.94
b,z
60.89
b,z
2.70 <0.001
FRE 400 24.57
x
33.39
xy
40.57
b,y
57.06
b,z
63.53
b,z
2.64 <0.001
FRE 800 22.47
w
34.21
x
45.62
b,y
60.23
b,z
65.69
b,z
2.02 <0.001
P- alue ea men 0.089 0.270 <0.001 <0.001 <0.001
a
SE 250, SE 500 and SE 1000 =250, 500 and 1000 mg all- ac-⊍- ocophe yl ace a e kg
−1
eed; RE 200, RE 400 and RE 800 =200, 400 and 800 mg ose-
ma y ex ac kg
−1
eed; FRE 200, FRE 400 and FRE 800 =200, 400 and 800 mg a embedded osema y ex ac kg
−1
eed.
a,b,c
Values wi hin a column wi h di e en supe sc ip a e significan ly di e en (P<0.05).
w,x,y,z
Values wi hin a ow wi h di e en supe sc ip a e sig-
nifican ly di e en (P<0.05). SEM: s anda d e o o he mean.
www.soci.o g LN Leal e al.
wileyonlinelib a y.com/js a © 2020 The Au ho s.
Jou nal o The Science o Food and Ag icul u e published by John Wiley & Sons L d on behal o Socie y o Chemical Indus y.
J Sci Food Ag ic 2020; 100: 2922–2931
2928
I
SO2
in LTL muscle a e p esen ed in Table 7. Mea discolo a ion and
O
2
sa u a ion we e significan ly a ec ed by s o age ime in all he
g oups (P<0.001). Besides ha , an in e ac ion be ween an ioxi-
dan supplemen a ion and s o age ime was ound o he
A
580
−A
630
and I
SO2
pa ame e s (P<0.001). A e 12 and 14 days
o s o age, he A
580
−A
630
and I
SO2
alues we e significan ly
highe in he i amin-E-supplemen ed lamb mea (0.25, 0.50, o
1.0 g kg
−1
eed) compa ed wi h he mea o he con ol and
osema y-supplemen ed lambs (RE and FRE). Fo mea discolo -
a ion (A
580
−A
630
), Rene e and Mazuel
47
p oposed a alue o
12.5 as he lowe limi o colou accep abili y. In he cu en
s udy, apa om he i amin-E-supplemen ed lambs ha had
A
580
−A
630
alues highe han 12.5 un il 14 days o display, he
o he ea men s only p esen ed accep able A
580
−A
630
alues
un il day 9 o s o age. Simila ly, LTL muscle om lambs supple-
men ed wi h i amin E p esen ed I
SO2
alues highe han 21.0
a e 14 days o s o age; in con as , in lambs ed he con ol o
he osema y-supplemen ed die s (RE and FRE), highe I
SO2
alues
han 21.0 we e only main ained o 7 days o s o age.
Las ly, due o he lack o esponse o bo h RE sou ces (RE and
FRE) on lipid oxida ion, colou s abili y, myoglobin oxida ion,
and mea discolo a ion pa ame e s when compa ed wi h he con-
ol g oup, i is no possible o d aw any conclusions ega ding he
e ficacy o p o ec ion in ended by a embedmen .
CONCLUSIONS
This s udy demons a ed ha eeding lambs a concen a e die
supplemen ed wi h all- ac-⊍- ocophe yl ace a e o 14 days
be o e slaugh e esul ed in a gene al imp o emen in mea oxi-
da i e and colou s abili ies. This s udy demons a es also ha ,
unlike i amin E, supplemen a ion o lamb die s wi h an RE (dos-
age and a embedmen ) had no e ec on lipid oxida ion o mea
colou s abili y o lambs du ing he 14 days o s o age unde e ail
condi ions.
ACKNOWLEDGEMENTS
We g a e ully acknowledge he suppo o F anco & Na a o
S.A. (Za agoza, Spain) and Me caza ogoza S.A. (Za agoza, Spain).
We also app ecia e he echnical assis ance and suppo wi h
he polyphenol analysis p o ided by P o esso Rosa Ma ia
Lamuela-Ra en ós, José Al a enga, and Paola Qui e (Uni e si y
o Ba celona, Spain).
CONFLICT OF INTERESTS
The au ho s’con ibu ions a e as ollows: LNL was he p incipal
in es iga o and con ibu ed o he s udy design, da a analyses,
and in e p e a ion o he findings and w o e he manusc ip ;
Table 7. E ec o die a y supplemen a ion
a
wi h all- ac-⊍- ocophe yl ace a e (SE), osema y ex ac (RE), and a -embedded osema y ex ac (FRE)
on mea discolo a ion (A
580
−A
630
and I
SO2
) in aw lamb mea s o ed in modified-a mosphe e packaging (70% O
2
:30% CO
2
) kep o 1, 7, 9, 12, and
14 days unde e ail condi ions
Days o display
P- alue displayI em 1 7 9 12 14 SEM
A
580
−A
630
Con ol 34.66
z
30.94
z
19.29
a,y
8.88
a,x
7.60
a,x
2.14 <0.001
SE 250 38.16
z
38.52
z
36.84
c,z
33.19
b,xy
29.48
b,x
0.99 <0.001
SE 500 35.98
z
36.97
z
34.54
bc,yz
31.05
b,xy
28.93
b,x
1.07 <0.001
SE 1000 37.50
z
36.96
z
35.35
bc,yz
32.94
b,xy
31.02
b,x
1.02 <0.001
RE 200 36.84
z
28.43
z
17.77
a,y
9.00
a,x
7.55
a,x
2.31 <0.001
RE 400 36.76
z
28.47
z
17.60
a,y
6.80
a,x
6.09
a,x
2.05 <0.001
RE 800 36.55
z
32.41
z
19.25
a,y
8.41
a,x
5.93
a,x
1.71 <0.001
FRE 200 38.36
z
29.19
yz
22.47
a,xy
13.42
a,wx
9.74
a,w
2.12 <0.001
FRE 400 34.42
z
31.18
yz
24.81
ab,y
10.86
a,x
7.61
a,x
2.07 <0.001
FRE 800 36.76
z
31.11
z
20.62
a,y
8.58
a,x
5.61
a,x
1.66 <0.001
P- alue ea men 0.138 0.124 <0.001 <0.001 <0.001
I
SO2
Con ol 19.28
yz
21.27
z
20.69
yz
15.32
abc,xy
10.62
a,x
1.38 <0.001
SE 250 18.64
x
21.54
y
21.46
y
21.25
bc,y
21.08
b,y
0.33 <0.001
SE 500 16.57
x
22.38
y
21.86
y
21.82
c,y
20.76
b,y
0.53 <0.001
SE 1000 17.44
x
21.80
y
21.85
y
21.67
c,y
21.77
b,y
0.34 <0.001
RE 200 17.99
yz
20.94
z
18.33
yz
12.51
a,xy
8.35
a,x
1.45 <0.001
RE 400 17.76
y
21.51
y
18.60
y
10.91
a,x
7.20
a,x
1.21 <0.001
RE 800 17.90
yz
21.56
z
18.79
yz
14.31
ab,xy
9.04
a,x
1.18 <0.001
FRE 200 19.07
yz
21.51
z
20.12
yz
15.65
abc,xy
12.15
a,x
1.28 <0.001
FRE 400 17.83
xy
21.03
y
19.76
y
16.36
abc,xy
12.26
a,x
1.51 <0.001
FRE 800 17.92
yz
21.36
z
20.92
z
14.25
abc,xy
10.11
a,x
1.06 <0.001
P- alue ea men 0.990 0.808 0.790 <0.001 <0.001
a
SE 250, SE 500, and SE 1000: 250, 500, and 1000 mg all- ac-⊍- ocophe yl ace a e kg
−1
eed; RE 200, RE 400, and RE 800: 200, 400, and 800 mg ose-
ma y ex ac kg
−1
eed; FRE 200, FRE 400, and FRE 800: 200, 400, and 800 mg a embedded osema y ex ac kg
−1
eed.
a,b,c
Values wi hin a column wi h di e en supe sc ip a e significan ly di e en (P<0.05).
w,x,y,z
Values wi hin a ow wi h di e en supe sc ip a e sig-
nifican ly di e en (P<0.05). SEM: s anda d e o o he mean.
Die a y i amin E and osema y ex ac s on mea quali y in lambs www.soci.o g
J Sci Food Ag ic 2020; 100: 2922–2931 © 2020 The Au ho s.
Jou nal o The Science o Food and Ag icul u e published by John Wiley & Sons L d on behal o Socie y o Chemical Indus y.
wileyonlinelib a y.com/js a
2929
JMB con ibu ed o he s udy design and da a collec ion; JM-T,
LAdH, and WHH con ibu ed o he s udy design and da a in e -
p e a ion; JAB and MB con ibu ed o he mea analysis and da a
in e p e a ion. All au ho s ead and app o ed he final e sion o
he manusc ip . WHH, JAB, and MB ha e no conflic o in e es s.
LNL, JMB, JM-T, and LAdH a e employed by T ouw Nu i ion, an
animal nu i ion company.
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J Sci Food Ag ic 2020; 100: 2922–2931
2930
