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Animal
Feed
Science
and
Technology
211
(2016)
75–83
Contents
lists
available
at
ScienceDirect
Animal
Feed
Science
and
Technology
journal
homepage:
www.elsevier.com/locate/anifeedsci
Effect
of
feeding
lactating
cows
with
ensiled
mixture
of
Moringa
oleifera,
wheat
hay
and
molasses,
on
digestibility
and
efficiency
of
milk
production
M.
Cohen-Zindera,
H.
Leibovichb,
Y.
Vakninc,
G.
Sagid,
A.
Shabtaya,
Y.
Ben-Meire,
M.
Nikbachate,
Y.
Portnike,
M.
Yishaya,
J.
Mirone,∗
aAgricultural
Research
Organization,
Beef
Cattle
Section,
Newe
Ya’ar,
P.O.
Box
1021,
Remat
Yishay
30095,
Israel
bResearch
and
Development
Haemek,
Israel
cDepartment
of
Natural
Resources,
Agricultural
Research
Organization,
P.O.
Box
6,
Bet
Dagan
50250,
Israel
dEden
Research
Farm,
Beit-Shean
Valley,
Israel
eDepartment
of
Ruminant
Science,
Agricultural
Research
Organization,
P.O.
Box
6,
Bet
Dagan
50250,
Israel
a
r
t
i
c
l
e
i
n
f
o
Article
history:
Received
13
July
2015
Received
in
revised
form
1
November
2015
Accepted
2
November
2015
Keywords:
Moringa
oleifera
silage
Digestibility
Milk
production
Milk
anti-oxidative
activity
Lactating
cows
a
b
s
t
r
a
c
t
Moringa
oleifera
Lam.
(MO)
seedlings,
sown
at
high
density
on
June
2014
and
grown
under
irrigation
in
Israel,
were
harvested
at
45
d
intervals
to
yield
a
total
biomass
of
35
t
dry
matter
(DM)
per
ha.
Based
on
a
preliminary
study
in
glass-silos,
fresh
harvested
MO
from
the
2nd
harvest
was
mixed
with
chopped
wheat
hay
and
sugar
cane
molasses
at
a
ratio,
of
370:540:90
on
DM
basis,
respectively,
and
ensiled
in
40
pressed
bales
(700
kg
each)
wrapped
with
stretch
polyethylene
film.
This
silage
was
included
in
the
total
mixed
ration
(TMR)
of
lactating
cows
at
a
level
of
180
g/kg
DM
as
wheat
silage
and
hay
substitute.
Perfor-
mance
and
digestion
experiment
was
conducted
with
two
groups
of
21
milking
cows
each,
fed
individually,
either
the
MO
TMR
or
the
control
ration.
Voluntary
DM
intake
of
cows,
fed
the
control
TMR,
tended
to
be
1.22%
higher
than
that
of
the
MO-fed
cows
(p
=
0.09).
In
vivo
digestibility
of
DM,
neutral
detergent
fiber
(NDF),
cellulose,
hemicelluloses
and
crude
pro-
tein
were
higher
in
the
control
cows
compared
with
the
MO-fed
group.
Notwithstanding,
higher
yields
of
milk
and
4%
fat
corrected
milk
(FCM)
by
1.91%
and
4.26%,
respectively,
were
observed
in
the
MO-fed
cows.
Milk
fat
content
was
2.34%
higher
and
milk
protein
content
2.28%
lower
in
the
MO-fed
cows
than
in
their
control
counterparts.
This
was
reflected
in
a
2.37%
higher
energy
corrected
milk
(ECM)
yield
and
3.27%
increase
in
production
effi-
ciency
(kg
ECM/kg
DM
intake)
of
the
MO-fed
cows
compared
with
the
control
ones.
Milk
of
the
MO-fed
cows
was
also
characterized
by
20%
more
anti-oxidative
activity
than
that
of
the
control
cows.
Body
weight
gain,
however,
was
similar
in
both
groups.
It
is
therefore
suggested
to
ensile
mixture
of
MO
with
soy
hulls
or
corn
grains
as
higher
digestible
solid
additive
for
feeding
lactating
dairy
cows.
©
2015
Elsevier
B.V.
All
rights
reserved.
Abbreviations:
ADF,
acid
detergent
fiber;
ADL,
acid
detergent
lignin;
BW,
body
weight;
CP,
crude
protein;
DM,
dry
matter;
DMI,
dry
matter
intake;
ECM,
energy
corrected
milk;
EE,
ether
extract;
EGCG,
equivalent
of
epicatechin
galate;
FCM,
4%
fat
corrected
milk;
IVDMD,
in-vitro
dry
matter
digestibility;
LDCL,
luminol-dependent
chemiluminescence;
MO,
Moringa
oleifera;
NDF,
neutral
detergent
fiber;
RFI,
residual
feed
intake;
TMR,
total
mixed
ration.
∗Corresponding
author.
E-mail
address:
jmiron@agri.gov.il
(J.
Miron).
http://dx.doi.org/10.1016/j.anifeedsci.2015.11.002
0377-8401/©
2015
Elsevier
B.V.
All
rights
reserved.
76
M.
Cohen-Zinder
et
al.
/
Animal
Feed
Science
and
Technology
211
(2016)
75–83
1.
Introduction
Moringa
oleifera
Lam.
(MO)
is
a
tropical
tree
reported
to
have
nutritional,
therapeutic
and
prophylactic
properties
(Reyes-
Sanchez
et
al.,
2006;
Moyo
et
al.,
2010).
MO
originated
from
the
Indian
subcontinent,
and
later
became
naturalized
in
tropical
and
subtropical
areas
around
the
world.
It
is
known
to
produce
high
leaf
mass
which
is
a
potential
high
quality
forage
source
for
ruminants
(Foidl
et
al.,
2001;
Sanchez-Machado
et
al.,
2010).
There
are
several
advantages
of
using
MO
foliage
as
ruminants
feed:
It
is
drought
resistance
and
has
the
ability
to
grow
on
poor
soils
(Foidl
et
al.,
2001);
It
produces
high
leaf
mass
within
a
short
period,
and
being
perennial
in
nature,
it
can
be
harvested
several
times
in
the
same
growing
season
(Mendieta-Araica
et
al.,
2011a,b);
MO
leaves
are
characterized
by
high
crude
protein
content,
adequate
amino
acid
profile,
high
level
of
vitamins
A,
B,
and
C
(Sanchez-Machado
et
al.,
2010;
Mendieta-Araica
et
al.,
2011a;
Sultana
et
al.,
2015),
and
high
amounts
of
polyphenols
resulting
in
elevated
antioxidative
activity
(Verma
et
al.,
2009).
Moreover,
MO
leaves
can
be
fed
fresh
or
dried,
and
after
drying
can
be
stored
for
long
periods
without
deterioration
in
nutritive
value
(Foidl
et
al.,
2001).
Several
research
studies
examined
the
effects
of
using
fresh
or
dried
hand-cut
MO
foliage
as
supplement
or
substitute
for
local
tropical
forage
or
concentrates
in
the
rations
of
lactating
ruminants.
Studies
with
low
producing
cows
showed
increase
in
milk
yield
in
response
to
feeding
dried
or
fresh
leaves
and
soft
twigs
of
MO
supplement
(Reyes-Sanchez
et
al.,
2006;
Mendieta-Araica
et
al.,
2011a).
Studies
with
lactating
goats
also
showed
an
increase
in
milk
yield
in
response
to
feeding
MO
leaves
as
a
sesame
or
concentrate
replacer
(Sultana
et
al.,
2015).
Despite
the
great
potential
of
using
MO
leaves
as
high
quality
animal
feed,
its
global
commercial
use
has
been
restricted
only
to
small
farm
holders
leaning
on
hand-cut
and
not
on
modern
machinery-based
forage
producers.
The
reasons
for
this
restricted
exploration
are:
MO
is
a
fast
growing
tree
producing
a
thick
trunk
and
branches
that
create
problems
for
commercial
machinery-based
harvesters
(e.g.
forage
harvester
or
forage
combine),
routinely
used
in
the
forage
production
industry;
MO
leaves
are
characterized
by
high
moisture
content
(150–200
g/kg
DM)
and
therefore
became
moldy
during
direct
ensiling,
and
needed
a
week
of
wilting
in
the
field
before
successful
ensiling
(Cohen-Zinder
et
al.,
submitted
for
publication).
To
overcome
these
obstacles,
a
new
genetic
variety
of
M.
oleifera
was
developed
by
Dr.
Yiftach
Vaknin
(ARO,
Israel)
during
10
years
of
selective
breeding
of
MO
germplasm
acquired
from
India
and
Western
Africa.
This
new
MO
variety
was
grown
for
the
first
time
in
the
Mediterranean
area
at
a
high
sowing
density
(160,000
seeds/Ha)
under
drip-irrigation,
to
produce
forage,
which
was
harvested
at
intervals
of
45
d
(four
harvests/year
between
July
to
November)
to
yield
a
total
forage
mass
of
35
t
DM
per
ha.
However,
the
residual
trunk
and
thick
branches
left
in
the
field
after
each
harvest
(at
20
cm
height
above
previous
harvest),
interfered
physically
with
direct-wilting
of
the
fresh
high
moisture
forage
(180–200
g/kg
DM)
in
the
field,
and
dictated
immediate
evacuation
of
the
fresh
forage
mass.
A
previous
study
that
used
pre-wilted
(a
week
in
the
field)
MO
silage
as
forage
source
for
lactating
cows
(at
a
level
of
130
g/kg
of
TMR
DM)
resulted
in
an
increase
in
milk
production
and
milk
antioxidative
activity
(Cohen-Zinder
et
al.,
submitted
for
publication).
However,
commercial
harvests
at
intervals
of
45
d
precludes
wilting
for
a
week
in
the
field,
and
dictates
either
fast
drying
of
the
fresh
MO
forage
outside
the
field
to
avoid
its
spoilage
(a
costly
process),
or
direct
ensiling
with
solid
feed
additives.
A
preliminary
glass-silo
study
showed
that
direct
ensiling
of
mixtures
of
fresh
MO
with
sugar
cane
molasses
and
either
chopped
wheat
hay
or
soy
hulls
(at
DM
ratio
of
370:90:540,
respectively)
were
the
best
combinations
that
ensured
successful
ensiling
(Cohen-Zinder
et
al.,
submitted
for
publication).
However,
the
effect
of
feeding
the
ensiled
mixture
MO
+
wheat
hay
+
molasses,
as
a
supplement
to
high
producing
lactating
cows,
on
intake,
digestibility
and
efficiency
of
milk
production
was
never
examined.
The
objective
of
the
current
study
was
to
examine
the
effects
of
feeding
the
new
variety
of
MO
ensiled
in
a
mixture
with
wheat
hay
and
molasses,
at
a
ratio
on
DM
basis
of
370:540:90,
respectively,
as
substitute
for
wheat
silage
and
hay
in
dairy
cow
ration,
on:
intake,
digestibility,
milk
yield,
milk
composition,
milk
production
efficiency,
and
milk
anti-oxidative
activity.
2.
Materials
and
methods
2.1.
Growth
conditions
of
M.
oleifera
and
its
ensiling
The
optimal
agronomical
growth
conditions
for
the
new
MO
variety,
used
as
a
forage
crop
in
this
study,
were
determined
in
a
preliminary
study
(Cohen-Zinder
et
al.,
submitted
for
publication).
Seeds
of
the
new
MO
variety
were
sown
on
June
15,
2014
at
density
of
160,000
seeds/Ha,
in
a
2-ha
commercial
field
in
Eden
Farm,
Israel
(longitude:
35◦E,
latitude:
32◦N).
The
plants
were
drip-irrigated
every
week,
up
to
sum
of
800
mm
during
the
growth
season
(June
to
November).
Four
mechanical
harvests
were
conducted
at
45
d
intervals,
at
20-cm
stubble
height
in
first
cut,
and
additional
7-cm
above
each
of
the
three
following
cuts.
In
each
harvest,
the
MO
forage
(including
leaves,
soft
twigs
and
thin
branches)
was
harvested
and
chopped
to
at
least
2-
to
3-cm
particle
size
using
a
John
Deere
combine
harvester
(9660
STS;
John
Deere,
Moline,
IL),
loaded
directly
into
trucks,
delivered
to
Nahalal
Feeding
Center
(29
miles
distance)
and
weighed
before
ensiling.
Forage
yield
of
the
entire
field
summarizing
the
four
harvests
of
2014
was
35
t
DM/ha.
A
preliminary
experiment
showed
that
direct
ensiling
of
a
mixtures
of
fresh
MO
with
sugar
cane
molasses
and
chopped
wheat
hay
or
soy
hulls
(at
DM
ratio
of
370:90:540
g/kg,
respectively)
were
the
best
combinations
that
ensured
successful
ensiling
expressed
in
less
than
50
g/kg
DM
loss,
low
silage
pH
(4.0)
and
above
300
g/kg
DM
content
(Cohen-Zinder
et
al.,
M.
Cohen-Zinder
et
al.
/
Animal
Feed
Science
and
Technology
211
(2016)
75–83
77
Table
1
Chemical
composition
of
the
main
forages
included
in
the
experimental
TMRs.
Composition
(g/kg
DM)
MO
silage1Wheat
hay
Wheat
silage
SEM
p-Value
pH
4.00
4.12
0.10
0.45
Lactate
55.0a36.2b1.55
0.04
Acetate
15.1
12.0
1.45
0.20
Propionate
0.55 0.60 0.15 0.65
DM
(g/kg
of
wet) 345b890a353b13.9
0.01
Organic
matter
879b908ab 914a4.14
0.05
Crude
protein
140a103b105b6.35
0.04
Neutral
detergent
fiber
571ab 591a559b4.81
0.10
Cellulose
329
325
323
5.90
0.34
Hemicellulose
182b218a195b4.25
0.03
Lignin
60.0a48.0b41.0c1.46
0.05
In
vitro
DM
digestibility20.62b0.63b0.67a0.01
0.01
1MO
silage
=
Moringa
oleifera
silage
composed
of
a
mixture
on
DM
basis
of:
370
g/kg
MO,
540
g/kg
chopped
wheat
hay
and
90
g/kg
sugar-cane
molasses,
ensiled
in
pressed
bales
wrapped
with
polyethylene
stretched
films.
2Composition
of
fresh
pre-ensiled
Moringa
oleifera
was:
DM,
180
g/kg;
ash,
93.8
g/kg
DM;
crude
protein,
173
g/kg
DM;
NDF,
476
g/kg
DM;
hemicellulose,
77
g/kg
DM;
cellulose,
345
g/kg
DM;
lignin,
54
g/kg
DM;
in
vitro
DM
digestibility,
0.57.
a,b,c Means
within
row
marked
with
different
superscripts
differ
significantly
at
p
<
0.05.
submitted
for
publication).
Sugarcane
molasses
were
introduced
before
the
ensiling
process
into
the
mixture
containing
MO
+
wheat
hay
or
soy
hulls,
because
the
ensiling
study
in
glass-silos
had
shown
that
MO
had
high
buffer
capacity
due
to
it’s
high
protein
and
soluble
phenolics
content,
and
therefore
without
molasses
addition
lactic-acid
fermentation
was
restricted
and
silage
pH
remained
higher
than
4.8.
Based
on
this
preliminary
study,
fresh
MO
forage
(DM
content,
180
g/kg)
from
the
2nd
harvest
(August
15,
2014)
was
pre-
mixed
for
10
min
in
a
mixing
wagon
(Lachish
IndustriesLTD,
Sderot,
Israel)
with
chopped
wheat
hay
and
sugar
cane
molasses
(at
DM
ratio
of
370:540:90,
respectively)
and
loaded
into
a
compression
chamber
by
a
conveyor
belt.
The
compressed
bales
were
then
lowered
on
to
a
rotating
tray
and
wrapped
with
8–9
layers
of
stretch
polyethylene
UV
resistant
film,
32
m
thickness
(Poleg
Industries,
Kibbutz
Gevim,
Israel)
as
previously
described
(Weinberg
et
al.,
2011).
Forty
MO
bales
of
about
700
kg
each
were
prepared
and
delivered
to
Bet-Dagan
dairy
barn
and
stored
(ensiled)
for
90
d
at
the
feeding
center
until
the
feeding
trial
began
at
November
2014.
Bales
were
opened
and
the
MO
silage
was
sampled
(four
bales
–
replicates)
from
the
middle
of
each
bale
before
it
was
mixed
in
the
MO
TMR.
The
MO
TMR
was
then
fed
to
lactating
cows
as
substitute
for
136
g/kg
DM
wheat
silage
and
44
g/kg
DM
wheat
hay
of
the
control
TMR
(Table
2).
The
wheat
hay
(variety
Galil)
mixed
with
MO
and
molasses
prior
to
the
ensiling
was
similar
to
the
one
used
in
the
feeding
trial,
and
harvested
at
the
early
stage
of
grain
maturation.
The
wheat
silage
of
the
control
TMR
(variety
Galil)
was
grown
in
a
near-by
field
and
harvested
at
the
end
of
the
soft-dough
stage
of
grains
maturation,
prior
to
ensiling
in
regular
cement
silo.
Chemical
composition
and
in
vitro
digestibility
of
the
samples
obtained
from
MO
silage,
wheat
silage
and
chopped
wheat
hay
(4
replicates
for
each
substrate)
are
shown
in
Table
1.
2.2.
Analytical
procedures
and
in
vitro
digestibility
Dry
matter
content
of
the
MO
silage,
wheat
hay
and
wheat
silage
used
in
the
TMRs,
TMR
samples,
orts,
and
dry
feces
samples
from
the
cows,
was
determined
by
drying
in
an
air-forced
oven
for
48
h
at
60 ◦C.
The
dry
samples
were
ground
through
a
Wiley
mill
(Philadelphia,
PA,
USA)
using
a
1-mm
screen
before
chemical
analyses.
Ash
was
determined
according
to
method
942.05
(AOAC
International,
2000).
Neutral
detergent
fiber
(NDF)
analyzed
with
heat-stable
amylase
and
without
Na-sulfite,
acid
detergent
fiber
(ADF)
and
acid
detergent
lignin
(ADL)
were
determined
according
to
the
sequential
method
of
Van
Soest
et
al.
(1991)
by
an
ANKOM
fiber
analyzer
(ANKOM220 Technology,
Macedon,
NY,
USA),
and
expressed
inclusive
of
residual
ash.
Hemicellulose
was
defined
as
NDF
−
ADF
and
cellulose
as
ADF
−
ADL.
Crude
protein
(CP)
was
determined
according
to
the
Kjeldahl
method
(procedure
14.068
in
AOAC,
1980).
Ether
extract
(EE)
was
determined
according
to
method
2003.05
(AOAC
International,
2000).
Water
extract
was
prepared
manually
(5
g
DM/100
g
water)
from
each
replicate
(four
replicates)
of
the
MO
+
wheat
+
molasses
silage
as
well
as
from
the
wheat
silage,
to
measure
pH,
lactate
and
volatile
fatty
acids.
Lactic
acid
in
the
water
extract
was
determined
(in
triplicate)
using
a
spectrophotometric
method
according
to
Barker
and
Summerson
(1941).
Volatile
fatty
acids
in
the
water
extract
were
determined
by
gas
chromatograph
equipped
with
a
semi-capillary
nitroterephthalic
acid-modified
polyethylene
glycol
column
(Hewlett
Packard,
Waldborn,
Germany)
over
a
temperature
range
of
40–230 ◦C.
The
pH
was
determined
in
each
water
extract
of
the
silages
with
a
pH-meter
(PL
600,
MRC,
Netanya,
Israel).
In
vitro
DM
digestibility
(IVDMD)
was
calculated
in
the
dried
ground
samples
according
to
the
two-stage
technique
of
Tilley
and
Terry
(1963),
using
rumen
fluid
obtained
from
two
ruminally
fistulated
dry
Holstein
cows,
each
sample
in
four
replicates.
The
rumen
fluid
donor-cows
were
fed
7
kg
(DM)
of
wheat
hay
and
3
kg
(DM)
of
total
mixed
ration
(TMR)
comprised
of
300
g/kg
grains,
350
g/kg
wheat
silage,
150
g/kg
soybean
meal,
and
200
g/kg
by-products
(cottonseed,
wheat
bran,
and
gluten
feed).
78
M.
Cohen-Zinder
et
al.
/
Animal
Feed
Science
and
Technology
211
(2016)
75–83
Table
2
Ingredients
of
the
total
mixed
rations
(TMR)
fed
to
lactating
cows.
Ingredients
(g/kg
TMR
DM)
Control
TMR
Moringa
oleifera
TMR
MO
silagea0
180
Wheat,
silage
209
73.0
Wheat
hay
(5–10
cm
length)
88.0
44.0
Vetch
hay
23.0
23.0
Corn
grain,
ground 294
294
Barley
grain
rolled 19.0 19.0
Soybean
meal
35.0
35.0
Ca-LCFAb10.0
10.0
Corn
gluten
feed
137
140
Urea
3.0
0
Dry
distillers
grains
(corn)
62.0
62.0
Rapeseed
meal
45.0
45.0
Non
linted
cotton
grain
20.0
20.0
Whey
32.0
32.0
Calcium
carbonate 7.00 7.00
Sodium
chloride
5.00
5.00
Sodium
bicarbonate
10.0
10.0
Minerals
+
vitamins
mixc1.00
1.00
aMO
silage
=
Moringa
oleifera
silage
composed
of
a
mixture
on
DM
basis
of:
370
g/kg
MO,
540
g/kg
chopped
wheat
hay
and
90
g/kg
sugar-cane
molasses,
ensiled
in
pressed
bales
wrapped
with
polyethylene
stretched
films.
bCalcium
salts
of
long-chain
fatty
acids.
cContaining
(g/kg
of
mix
DM):
Zn,
24;
Fe,
24;
Cu,
12.8;
Mn,
24;
I,
1.44;
Co,
0.32;
Se,
0.32;
16,000,000
IU
of
vitamin
A;
3,200,000
IU
of
vitamin
D3;
and
48,000
IU
of
vitamin
E.
Table
3
Chemical
composition
of
the
two
TMRs
fed
to
lactating
cows.
Composition
(g/kg
DM
TMR)
Control
TMR
Moringa
oleifera
TMR
SEM
p
Dry
matter
(g/kg
wet
TMR)
634
600
9.90
0.12
Organic
matter
924
925
4.10
0.78
Crude
protein 165
165
2.00
0.89
Ether
extract
52.5
50.0
1.65
0.32
Neutral
detergent
fiber
(NDF)
333
348
3.18
0.04
Forage
NDF
180
180
3.20
0.76
Hemicellulose
176
164
6.10
0.34
Cellulose
120
138
3.56
0.03
Lignin
30.4
37.2
2.00
0.02
Non
structural
carbohydratea373
361
4.12
0.10
In
vitro
DM
digestibility
0.75
0.72
0.007
0.01
aNon
structural
carbohydrate
fraction
was
calculated
as
=
organic
matter
−
NDF
−
crude
protein
−
ether
extract.
2.3.
Intake
and
performance
in
lactating
cows
The
forty
bales
of
MO
silage
were
transferred
to
Yavne
Feeding
Center
(Kibutz
Yavne,
Israel)
and
used
for
daily
fresh
preparation
of
the
MO-TMR.
The
freshly
prepared
MO
and
control
TMRs,
were
delivered
to
the
Agricultural
Research
Orga-
nization
dairy
barn
in
Bet-Dagan
(Israel),
and
fed
to
one
of
the
two
groups
of
21
lactating
cows,
each.
Composition
of
the
ingredients
included
in
the
two
TMRs
is
presented
in
Table
2
and
the
chemical
composition
of
the
two
TMRs
is
presented
in
Table
3.
The
two
groups
of
cows
were
similar
at
the
onset
of
the
experiment
(Mean
±
SE)
in
lactation
number
(3.3
±
0.3),
days
in
milking
(192
±
7.0)
and
milk
yield
(45.3
±
0.96).
Cows
were
blocked
by
DIM
(60–120
vs.
120–180)
and
daily
milk
production
(35–40
vs.
40–50
kg/cow)
and
randomly
assigned
within
block
to
pairs
fed
the
two
dietary
treatments.
The
two
groups
were
fed
the
two
TMRs
(control
vs.
MO)
for
two
weeks
of
adaptation,
followed
by
six
weeks
of
experiment.
Since
in
the
ARO
dairy
barn
each
cow
uses
an
individual
feeder,
the
TMRs
were
fed
only
once
a
day
at
1000
h
ad
libitum,
allow-
ing
for
50–100
g/kg
orts.
Cows
were
milked
3
times
daily
at
0600,
1400,
and
2200
h.
The
cows
were
fed
individually
via
a
computerized
monitoring
system
designed
to
electronically
identify
them
individually
and
to
automatically
record
their
daily
feed
intake
(Miron
et
al.,
2003).
Daily
DMI
of
individual
cows
was
determined
based
on
DM
content
in
TMR
sampled
daily
and
feed
refusals.
Body
weight
(BW)
data
were
recorded
by
an
automatic
walk-over
scale
3
times
per
day
while
cows
were
entering
the
milking
parlor.
Changes
in
BW
were
calculated
as
the
difference
in
weight
between
the
week
before
the
onset
of
the
experiment
and
at
week
6
of
the
experiment.
Milk
yield
(kg)
and
content
of
milk
fat,
protein,
and
lactose,
were
recorded
daily
for
each
cow
by
an
automatic
meter
equipped
with
on-line
near-infra-red-spectroscopy
measurement
(Afilab,
Afimilk
Ltd.,
Kibbutz
Afikim,
Israel).
In
parallel
for
validation
of
the
on-line
milk
composition
measurement,
milk
samples
were
collected
in
3
sequential
milking’s
on
a
weekly
basis
throughout
the
study,
stored
at
4◦C
in
the
presence
of
2-bromo-
2-nitropropane-1,3-diol,
until
infrared
analysis
for
fat,
protein,
urea
and
lactose
content,
using
a
MilkoScan
4000
analyzer
M.
Cohen-Zinder
et
al.
/
Animal
Feed
Science
and
Technology
211
(2016)
75–83
79
(Foss
Electric
A/S,
Hillerød,
Denmark).
Energy
corrected
milk
(ECM)
yield
was
calculated
using
the
following
equation
(NRC,
2001):
ECM
(kg/d)
=
milk
yield
(kg/d)
×
{[0.3887
×
milk
fat
(%)]
+
[0.2356
×
milk
protein
−
urea
(%)]
+
[0.1653
×
milk
lactose
(%)]}/3.1338
MJ/kg.
Residual
feed
intake
(RFI)
used
to
examine
feeding
efficiency
was
calculated
on
a
daily
basis
as:
RFI
(kg
DM/d)
=
DMI
(kg
DM/d)
−
predicted
DMI
(which
was
calculated
according
to
the
NRC
(2001)
equation:
predicted
DMI
(kg/d)
=
(FCM*0.372
+
BW0.75*0.0968*(1
−
e(0.192*(week
in
lactation+3.67)).
The
animal
performance
and
digestibility
study
was
carried
out
according
to
the
guidelines
and
under
the
supervision
of
the
Agricultural
Research
Organization
Animal
Care
Committee.
2.4.
In
vivo
digestibility
measurements
in
lactating
cows
At
wk
5
of
the
experiment,
4
d
were
assigned
for
daily
sampling
of
TMR,
feed
refusals,
and
feces
from
10
cows
selected
from
each
group.
These
cows
were
selected
in
pairs
to
create
two
sub-groups
of
the
dietary
treatment
similar
in
milk
yield
(44.0
kg/d).
Estimation
of
daily
fecal
excretion
was
based
on
analyses
of
indigestible
NDF
concentrations
in
feces
and
the
TMRs.
Samples
of
the
TMR
and
refusals
were
composited
on
a
daily
basis,
dried
(60 ◦C
for
48
h),
and
ground
through
a
1-mm
sieve
(Wiley
Mill,
Philadelphia,
PA,
USA).
Fecal
grab
samples
were
collected
3
times
daily
for
4
d,
at
8-h
intervals,
each
day
2.5
h
later
than
the
preceding
day.
Fecal
samples
were
then
composited
(on
a
DM
basis)
from
each
cow
during
the
collection
period,
dried
at
60 ◦C
for
48
h
in
a
forced-air
oven,
ground
to
pass
through
1-mm
sieve,
and
then
used
for
chemical
analyses
as
described
above,
and
for
analysis
of
indigestible
NDF
content
(as
internal
marker)
in
the
TMR
and
fecal
pooled
samples.
The
two-stage
in
vitro
digestibility
technique
of
Tilley
and
Terry
(1963)
was
used
to
analyze
the
content
of
residual
indigestible
NDF
in
the
TMR
and
in
the
pooled
fecal
samples
of
each
cow,
after
incubation
with
rumen
fluid
for
72
h
followed
by
incubation
for
48
h
with
HCl-pepsin
(Adin
et
al.,
2009).
The
quantitative
ratio
indigestible
NDF
in
TMR/indigestible
NDF
in
feces
is
identical
to
the
ratio
Fecal
DM/DMI
of
each
cow,
which
is
actually
the
reciprocal
of
in
vivo
DM
digestibility
according
to
the
following
equation
(Adin
et
al.,
2009):
DM
digestibility
=
1
−
(TMR
indigestible
NDF/fecal
indigestible
NDF).
The
digestibility
values
of
each
chemical
component
(i.e.,
DM,
CP,
NDF,
cellulose,
and
hemicellulose)
were
calculated
for
every
cow
using
its’
proportion
between
intake
and
fecal
output
according
to
this
equation
(Adin
et
al.,
2009).
2.5.
Milk
anti-oxidative
activity
measurement
Luminol-enhanced
chemiluminescence
assay
(Ginsburg
et
al.,
2005)
was
used
to
measure
the
reducing
antioxidant
poten-
tial
of
milk.
Briefly,
individual
milk
samples
(20
l
from
each
sampling
time)
were
added
to
a
reaction
mixture
containing
10
l
luminol
(10
M),
10
l
morpholinosydnonimine
(SIN-1,
1
mM),
20
l
sodium
selenite
(2
mM)
and
10
l
Co2+ (SIN-1
cocktail).
This
radical-generating
cocktail
simultaneously
generates
a
flux
of
peroxide
and
NO.
The
capacity
of
milk
samples
to
quench
the
luminol-enhanced
chemiluminescence
generated
by
the
cocktail
was
measured
in
a
LUMAC/3M
Biocounter
M2010
connected
to
a
linear
recorder.
The
resulting
light
output
was
recorded
as
counts/min.
for
6
min.
The
luminol-
dependent
chemiluminescence
(LDCL)
values
were
expressed
as
equivalent
of
epicatechin
galate
(EGCG,
M),
using
a
calibration
curve
in
which
EGCG
was
plotted
against
LDCL.
The
antioxidative
potential
of
the
milk
was
expressed,
each
minute,
relative
to
the
SIN-1
cocktail.
2.6.
Rumen
pH
measurement
Ten
cows
from
each
group
(same
cows
used
for
digestibility
measurements)
were
used
every
day
during
week
6
for
rumen
fluid
sampling.
During
this
week
cows
were
fed
once
a
day
at
0600
for
one
hour,
following
by
feed
prevention
for
six
hours
and
then
allowed
to
eat
for
ad-lib
intake.
This
feeding
routine
was
chosen
since
preliminary
observations
have
demonstrated
maximal
and
minimal
rumen
pH
at
1
h
pre-feeding
and
6
h
post
feeding,
respectively.
Five-hundred
ml
of
rumen
fluid
samples
free
of
saliva
were
collected
from
each
cow
with
a
rumen
vacuum
sampler
at
1
h
pre-feeding,
6
h
after
feeding,
12
h
post
feeding
and
18
h
post
feeding.
The
vacuum
pump
was
turned
on
only
after
the
metal
coated
sampler
pipe
was
inserted
through
the
esophagus
and
located
in
the
math
phase
of
the
rumen,
to
avoid
saliva
contamination.
The
rumen
pH
values
were
immediately
determined
by
a
portable
pH-meter
(PL
600,
MRC
Israel),
and
are
presented
in
Table
4.
2.7.
Statistical
analysis
Differences
between
the
three
roughages
and
the
two
types
of
TMR
fed
to
21
pairs
of
lactating
cows
with
respect
to
chemical
composition
(Tables
1
and
3),
in
vivo
digestibility
and
rumen
pH
(Table
4),
were
tested
for
significance
by
F-test
using
JMP-5
software
(SAS,
1996).
Tukey–Kramer
test
(SAS,
1996)
was
used
for
comparison
between
means.
Daily
DM
intake,
BW,
milk
yield,
milk
fat,
protein
and
lactose
content,
ECM
yield,
and
production
efficiency
(ECM/DMI
or
RFI)
data
(Table
4)
were
averaged
by
cow
for
the
42
d
in
the
experiment
(repeated
measures
for
each
cow).
Data
were
analyzed
as
a
randomized
complete
block
design
using
the
mixed
model
of
SAS
software
(SAS,
1996).
The
statistical
model
included
the
effects
of
dietary
treatment,
block,
and
cow
by
block
interaction.
Cow
within
block
was
delineated
as
the
random
80
M.
Cohen-Zinder
et
al.
/
Animal
Feed
Science
and
Technology
211
(2016)
75–83
Table
4
Rumen
pH
and
in
vivo
digestibility
of
the
two
total
mixed
rations
(TMR)
fed
to
lactating
cows.
Criteria
Control
TMR
Moringa
oleifera
TMR
SEM
p
N
10
10
1
h
pre-feeding
rumen
pH
7.02
7.02
0.09
0.65
6
h
post-feeding
rumen
pH
6.21
6.31
0.03
0.10
Average
rumen
pH
(4
samples/d) 6.59 6.68 0.06 0.21
In
vivo
digestibility
Dry
matter
0.723
0.677
0.007
0.02
Neutral
detergent
fiber
0.519
0.473
0.008
0.02
Cellulose
0.604
0.566
0.001
0.05
Hemicellulose
0.574
0.491
0.001
0.01
Crude
protein
0.707
0.661
0.001
0.05
effect
in
the
model
and
used
to
test
treatment
and
block
effects,
whereas
repeated
measurement
of
daily
data
were
tested
for
the
residual
error.
3.
Results
3.1.
Composition
and
in
vitro
digestibility
of
forages
and
TMRs
The
mixture
of
MO
silage
was
characterized
by
acidic
pH
(4.06),
and
DM
content
of
345
g/kg,
similar
to
that
of
the
wheat
silage
(Table
1).
The
MO
silage
contained
more
protein
than
the
wheat
silage
or
hay
and
therefore
urea
was
added
to
the
control
TMR
to
create
equal
CP
content
in
both
TMRs
(Table
2).
The
in
vitro
DM
digestibility
of
wheat
silage
was
8.04%
higher
than
that
of
the
MO
silage.
This
study
used
180
g/kg
MO
silage
in
the
MO-TMR
(on
DM
basis)
as
substitute
for
136
g/kg
wheat
silage
and
44
g/kg
wheat
hay
included
in
the
control
TMR.
Chemical
composition
data
shows
higher
content
of
NDF,
cellulose
and
lignin
as
well
as
lower
IVDMD
of
the
MO
TMR
compared
with
the
control
TMR
(Table
3)
in
accordance
with
the
differences
in
IVDMD
between
the
MO
silage
and
wheat
silage
(Table
1).
3.2.
Intake,
digestibility
and
milking
performance
in
cows
In
vivo
digestibility
data
is
shown
in
Table
4.
Cows
fed
with
the
control
TMR
tended
to
ingest
1.22%
more
DM
(p
=
0.09)
than
the
control
cows
(Table
5).
This
advantage
in
intake
might
be
related
to
the
lower
NDF
content
and
higher
in
vivo
digestibility
of
DM
and
NDF
found
in
the
control
cows
compared
with
the
MO
group
(Table
4).
Cows
fed
with
the
control
TMR
ingested
3.2%
less
NDF
but
had
a
9.7%
advantage
in
NDF
digestibility
and
6.96%
in
CP
digestibility,
compared
with
the
MO-fed
cows
(Table
4).
Average
daily
rumen
pH
values
were
similar
and
relatively
high
in
cows
fed
the
two
TMRs.
The
lactation
performance
data
of
cows
fed
with
the
two
dietary
treatments
is
presented
in
Table
5.
Although
the
control
cows
ingested
8.43%
more
digestible
DM/d
than
the
MO
group
(18.0
kg
and
16.6
kg
digestible
DM/d
respectively)
it
was
not
reflected
in
milk
production
which
turned
to
be
higher
in
the
MO
group
(p
=
0.01).
Milk
fat
content
was
also
higher
in
cows
fed
with
the
MO
diet,
while
milk
protein
content
was
higher
in
the
control
cows
(p
<
0.01).
This
was
reflected
in
a
2.37%
higher
ECM
yield
of
the
MO-fed
cows
compared
with
the
control
group.
The
lower
DM
intake
and
the
higher
ECM
yield
of
the
MO-fed
cows
resulted
in
a
3.27%
increase
in
their
ECM
production
efficiency
(ECM/DMI)
compared
with
the
control
Table
5
Intake
and
performance
in
lactating
cows
fed
the
two
total
mixed
rations
(TMR).
Parameter
Control
TMR
Moringa
oleifera
TMR
SEM
p
N
21
21
Dry
matter
intake
(DMI,
kg/cow/d)
24.9
24.6
0.08
0.09
Milk
yield
(kg/cow/d)
41.8
42.6
0.13
0.01
Milk
fat
(g/kg)
34.1
34.9
0.10
0.01
Milk
protein
(g/kg)
31.4
30.7
0.10
0.01
Milk
lactose
(g/kg)
48.9
48.5
0.10
0.01
4%
FCMayield
(kg/cow/d)
35.2
36.7
0.10
0.01
ECMayield
(kg/cow/d)
38.0
38.9
0.09
0.01
Efficiency
(kg
ECMa/kg
DMI)
1.53
1.58
0.01
0.01
RFI
(kg
DMI
–
predicted
kg
DMI)b−0.71 −1.52
0.08
0.01
Milk
AOA
on
week
0
(EGCG
M)c2.61
2.57
0.22
0.88
Milk
AOA
on
week
4
(EGCG
M)c2.63
3.12
0.10
0.01
Body
weight
change
(kg/cow/d)
0.35
0.34
0.09
0.97
aFCM
=
4%
fat
corrected
milk;
ECM
=
energy
corrected
milk.
bRFI
=
residual
feed
intake
calculated
as
the
actual
DMI
–
predicted
DMI
[according
to
the
equation
of
NRC,
2001].
cAOA
=
antioxidative
activity
in
milk
expressed
as
equivalent
of
epicatechin
galate
(EGCG,
M).
M.
Cohen-Zinder
et
al.
/
Animal
Feed
Science
and
Technology
211
(2016)
75–83
81
group.
RFI
data
of
the
two
groups
also
showed
an
advantage
in
efficiency
toward
the
MO-fed
cows
which
consumed
1.52
kg
less
DM/d
than
expected
according
to
NRC
(2001)
model.
The
control
cows
were
less
efficient
with
this
respect.
Both
groups
showed
similar
BW
gain
during
the
6
weeks
of
experimental
period.
Antioxidant
activity
in
milk
was
slightly
elevated
by
1%
in
the
control
group
during
the
4
weeks
of
the
experiment
(Table
5).
However,
antioxidant
activity
in
milk
of
the
MO-fed
cows
in
the
4th
wk
of
the
experiment
increased
by
21%
compared
to
the
activity
at
the
onset
of
the
experiment
(p
=
0.01).
Thus,
actual
advantage
of
20%
in
milk
antioxidant
activity
was
found
in
the
MO-fed
cows
compared
with
the
control
cows.
4.
Discussion
This
study
used
for
the
first
time
M.
oleifera
Lam.
(MO)
grown
under
drip-irrigation
in
the
Mediterranean
area
and
harvested
by
commercial
combine
at
45
d
intervals
to
yield
a
total
biomass
of
35
t
dry
matter
(DM)
per
ha.
This
study
also
used
for
the
first
time
MO
ensiled
with
solid
feed-stuffs
and
used
as
a
feed
additive
for
lactating
cows.
Performance
data
of
this
study
shows
that
yields
of
milk,
FCM
and
ECM
were
significantly
higher
by
0.8,
1.5
and
0.9
kg/d,
respectively,
in
cows
fed
MO-silage
compared
to
the
control
group.
These
findings
are
in
accordance
with
previous
studies
which
used
fresh
or
dried
MO
foliage
as
substitute
for
tropical
forage
or
forage
+
concentrates
in
rations
of
lactating
ruminants.
Reyes-Sanchez
et
al.
(2006)
showed
that
milk
yield
was
increased
in
dairy
cows
by
58%
and
65%
when
fed
supplement
of
2
and
3
kg
dried
MO
leaves
per
day,
respectively,
as
substitute
for
local
forage.
In
another
experiment
with
cross-bred
low
yield
dairy
cows,
dried
MO
leaf
meal
substituted
430,
730
and
1000
g/kg
of
cotton
seed
cake
in
concentrated
mixtures
while
significantly
increasing
milk
yield
(Mendieta-Araica
et
al.,
2011a).
Mendieta-Araica
et
al.
(2011b)
tested
the
effect
of
feeding
low
yielding
dairy
cows
with
diets
based
mainly
on
either
fresh
MO
foliage
plus
90
g/kg
molasses
or
600
g/kg
elephant
grass
plus
400
g/kg
concentrate
or
ensiled
MO
foliage
with
90
g/kg
molasses.
They
found
higher
intake
and
digestibility
in
cows
fed
MO
silage
compared
with
the
other
two
diets.
But,
the
differences
among
the
three
groups
in
milk
yield
(13.6–13.9
kg/d)
and
milk
composition
were
not
significant.
Sultana
et
al.
(2015)
found
higher
milk
and
ECM
yields
in
lactating
goats
fed
dried
MO
leaves
as
substitute
for
sesame
meal
or
concentrates
in
the
diet.
It
should
be
noted,
however,
that
all
of
these
previous
studies
performed
on
lactating
ruminants,
used
MO
leaves
and
soft
twigs
which
were
grown
on
trees,
hand
–
harvested
and
then
dried
or
ensiled
in
a
small
scale.
Thus,
DM
digestibility
of
the
MO
foliage
in
these
studies
was
much
higher
(>0.70)
than
that
of
the
commercially
harvested
MO
of
the
current
study
(0.57
IVDMD,
Table
1)
which
includes
leaves,
soft
twigs
and
small
branches
and
therefore
contained
high
levels
of
NDF,
cellulose
and
lignin
(Table
1).
Since,
the
cutting
height
used
for
the
Moringa
in
this
study
is
responsible
for
these
compositional
and
digestibility
changes,
it
should
be
considered
when
harnessing
MO
for
livestock
feeding
or
for
future
research.
In
addition,
the
cows
used
in
the
previous
studies
were
low
milk
producers
(up
to
14
kg/d)
and
fed
a
restricted
amount
of
DMI
(up
to
11
kg
DM/d).
Unfortunately,
there
is
lack
of
knowledge
in
the
literature
about
the
effects
of
feeding
MO
forage
(including
leaves,
soft
twigs
and
branches)
harvested
and
chopped
by
commercial
combine
and
ensiled
in
large
scale,
on
intake,
digestibility
and
efficiency
of
milk
production
in
high
producing
cows
as
demonstrated
in
the
current
study.
Possible
explanation
to
the
higher
milk,
FCM
and
ECM
yields
of
the
MO-fed
cows
compared
with
the
control
group
(Table
5),
might
be
related
to
the
high
anti-oxidative
activity
of
the
MO
silage
due
to
the
high
phenolic
components
of
the
MO
leaves
(Cohen-Zinder
et
al.,
submitted
for
publication;
Verma
et
al.,
2009),
which
were
shown
to
have
beneficial
effects
in
productive
ruminants
(Makkar,
2003;
Cohen-Zinder
et
al.,
submitted
for
publication).
Previous
studies
showed
that
moderate
concentrations
of
soluble
phenolics
and
tannins
in
the
diet
(20–40
g/kg
DM)
improved
production
efficiency
of
ruminants,
without
increasing
DM
intake,
as
manifested
by
increased
wool
growth,
BW
gain,
milk
yield
and
ovulation
rate
(Aerts
et
al.,
1999).
A
similar
trend
was
observed
in
a
previous
study
performed
on
lactating
cows,
fed
with
an
additive
of
40
g/kg
DM
concentrated
extract
of
pomegranate
pulp,
which
resulted
in
6.4%
increase
in
milk
production
of
early
lactation
cows
(Shabtay
et
al.,
2012).
In
another
study,
a
low
level
of
soluble
phenolics
addition
supplied
from
pomegranate
pulp
to
cows’
TMR,
resulted
in
improved
fat
yield
and
ECM
production
efficiency
compared
with
a
control
TMR.
This
advantage
was
attributed
to
25%
inhibition
of
in
vitro
methane
production
by
ruminal
methanogenic
bacteria
in
the
cows
fed
soluble
phenolics
(Shaani
et
al.,
2016).
Reduction
of
methanogenesis
by
phenolics
in
the
rumen
might
save
energy
which
otherwise
is
wasted
as