Ileal digestibility of amino acids of unheated and autoclaved

pea protein concentrate in broilers1

M. Frikha, D. G. Valencia,2 A. de Coca-Sinova,3 R. Lázaro, and G. G. Mateos4

Departamento de Producción Animal, Universidad Politécnica de Madrid,

Ciudad Universitaria, 28040 Madrid, Spain

ABSTRACT The effects of autoclaving 2 varieties of CP (87.6 vs. 82.2%) and most indispensable AA (e.g.,
micronized (fine grinding) pea protein concentrate 92.1 vs. 88.8% for Lys and 83.6 vs. 76.5% for Thr) of
(PPC) on the ileal digestibility (ID) of CP and amino the PPC. The improvement in CP and AA digestibility
acids (AA) were studied in broilers. There was a control with autoclaving varied with the PPC used and was
diet based on fermented soybean meal (FSBM) and 4 consistent with the reduction in TIA observed (9.4 to
extra diets in which the FSBM was substituted on a CP 2.8 mg/g for PPC-1 vs. 9.1 to 5.3 mg/g for PPC-2).
basis by PPC from 2 different pea cultivars (PPC-1 and The standardized ID (SID) of most indispensable AA
PPC-2), either unheated or autoclaved. Chicks were fed was similar for the FSBM and the PPC-2 and higher
a common diet from 1 to 17 d of age and, then, their for both than for the PPC-1 (P < 0.05). For Lys, the
respective experimental diets from 18 to 21 d of age. lowest SID value was observed for the FSBM and the
Each treatment was replicated 6 times. Autoclaving re- highest for the PPC-2 either unheated or autoclaved.
duced trypsin inhibitor activity (TIA) but had little It is concluded that the ID of the AA of the PPC im-
effect on the saponin content of the PPC. The apparent proved with heating and was in general higher for the
ID (AID) of CP was similar for the FSBM and the un- autoclaved PPC than for the FSBM. Consequently,
heated PPC and lower for both than for the autoclaved heat processed PPC is a good alternative to FSBM and
PPC. Autoclaving improved (P < 0.001) the AID of unheated PPC in starter diets for broilers.
Key words: amino acid digestibility, autoclaving, broiler, fermented soybean meal, pea protein concentrate
2013 Poultry Science 92:1848–1857
http://dx.doi.org/10.3382/ps.2013-03007

INTRODUCTION Different processes have been proposed to improve

the nutritional value of peas for poultry. Dehulling
Peas (Pisum sativum L.) are widely used in poultry reduces the content of crude fiber as well as that of
diets because of their high CP, Lys, and energy content some ANF present in the hull, such as tannins. How-
(Castell et al., 1996). However, the level of inclusion of ever, trypsin inhibitors (TI), lectins, and saponins are
peas in diets for young broilers is limited because of its located for the most part in the protein bodies of the
antinutritional factor (ANF) content, mainly protease cotyledon, and therefore, they are not removed by de-
inhibitors (Liener, 1989) and oligosaccharides (Igbasan hulling (Wiryawan and Dingle, 1999). Micronization
et al., 1997), which interfere with digestive processes (very fine grinding) of dehulled peas, followed by air
and reduce growth performance (Gatel and Grosjean, classification, allows the separation of the light (pro-
1990; Huisman et al., 1992). Also, peas contain an ap- tein) from the heavy (starch) fraction. The fine fraction
preciable amount of saponins, a cianogenetic glycosid- contains variable amounts of CP (40 to 60.5%) and
ic compound that causes bitterness, astringency, and residual starch (2.0 to 7.5%) and is commercially avail-
off-flavor and might reduce feed intake in poultry and able as pea protein concentrate (PPC; Valencia et al.,
other species (Price et al., 1985; McNeill et al., 2004). 2008). However, many of the ANF, including TI, con-
centrate in the protein fraction of the seed, which could
reduce growth performance when PPC is incorporated
©2013 Poultry Science Association Inc.
into broiler diets.
Received January 2, 2013.
Accepted March 19, 2013. Autoclaving consists in applying heat under certain
1 Funds from Project AGL2011-03506. Ministerio de Ciencia e In- pressure conditions during determined periods of time
novación, C.P. 28040 Madrid, Spain. to reduce the TI, lectins, and other heat-labile ANF
2 Current address: Nutral S.A., 28770 Madrid, Spain.
3 Current address: Urcacyl, 47007 Valladolid, Spain. contained in the legumes, improving their nutritive
4 Corresponding author: [email protected] value (Wiryawan and Dingle, 1999). However, saponins

1848
 PEA PROTEIN CONCENTRATE IN BROILERS 1849
are heat-stable (Liener, 2000), although some structural experiment. The trial was conducted as a completely
changes might happen with heating (Heng et al., 2006). randomized design with 5 dietary treatments consisting
Fermentation, together with the addition of selected of a control diet that used FSBM as the unique source
enzymes to defatted and decorticated soybean meal of protein and 4 additional diets arranged factorially
(SBM), removes to a high extent the soluble carbo- in which the FSBM was substituted on a CP basis by
hydrates and allergenic proteins contained in the meal, 1 of the 2 sources of PPC (PPC-1 and PPC-2), either
resulting in a soy product (fermented SBM, FSBM) unheated or autoclaved. In addition, a N-free diet was
that is low in fiber, lectins, TI, and oligosaccharides formulated and fed to 6 additional cages of broilers to
(Lenehan et al., 2007; Berrocoso et al., 2013). There- estimate the basal endogenous losses for the determina-
fore, the use of FSBM in substitution of SBM should tion of standardized ID (SID) of nutrients.
improve nutrient digestibility and growth performance
in young nonruminant animals. In this respect, FSBM
Ingredient and Diets
has been used successfully in starter diets for broilers
(González-Alvarado et al., 2007). The FSBM used was obtained from a conventional
Mariscal-Landín et al. (2002) compared the appar- trader and was produced from defatted and dehulled
ent ileal digestibility (AID) of amino acids (AA) of soya flakes via fermentation and enzymatic degrada-
unheated and heat-processed (HP) peas in pigs and re- tion, and contained by duplicate analysis 52.4% CP,
ported that heating improved considerably (up to 17%) 0.2 mg TI activity (TIA)/g, and 0.1 g of saponins/kg.
the digestibility of CP and of all AA (e.g., 77.6 vs. Two varieties of yellow peas (Pisum sativum, L.) with
87.6% for Lys). To our knowledge, no studies have been similar CP and TIA contents, were obtained at differ-
conducted comparing the ileal digestibility (ID) of the ent locations and time from a local supplier (Esasa,
AA of FSBM, unheated PPC, and HP PPC in broilers. Valladolid, Spain), cleaned, dehulled, and ground to a
The hypothesis tested in this study was that microniza- mean particle size of approximately 30 μm by using an
tion (very fine grinding) of unheated PPC could result impact mill (Circoplex mill 200 ZPS, Hosokawa-Alpine,
in ID of AA in broilers similar to those of FSBM and Augsburg, Germany). The finely ground peas were
that HP of the micronized PPC could increase further then passed through an air classifier equipment (Tur-
AA digestibility. The current experiment examined the boplex ATP 200, Hosokawa-Alpine) to separate the fine
influence of autoclaving 2 varieties of PPC on the ID and coarse fractions. The fine fraction (approximately
of AA in broilers. 28% of the original dehulled peas) corresponded to the
protein-rich fraction and contained by analysis 45.6 and
MATERIALS AND METHODS 44.6% CP, 9.4 and 9.1 mg of TIA/g, and 1.4 and 1.1
g of saponins/kg for PPC-1 and PPC-2, respectively.
The experimental procedures described in this re- The 2 varieties were split into 2 portions. The first
search were approved by the animal ethics committee portion of each variety was used as such, whereas the
of the Universidad Politécnica de Madrid and were in other portion was processed in a steam-jacketed auto-
compliance with the Spanish Guidelines for the Care clave before being included in the diets. The process
and Use of Animals in Research (Boletín Oficial Del consisted of heating the meal in a laboratory autoclave
Estado, 2007). (AES-75, Raypa, Terrassa, Spain) with a capacity of
75 L and a maximum pressure of 3.0 bars. The tank of
Bird Management, Husbandry, the autoclave was filled with distilled water up to the
and Experimental Design lower grid level. Individual batches (n = 10) of both
PPC varieties, weighing 2 kg each, were placed into
In total, 216 one-day-old straight-run broiler chicks the imperforated stainless-steel basket (38 cm diameter
(Cobb 500) with an initial BW of 40.4 ± 2.7 g were and 26 cm width) and autoclaved using a wet-cycle
used in this experiment. After arrival at the experi- program at 108°C for 8 min at 0.6 bars. The tempera-
mental station, the birds were weighed individually ture applied in this research (i.e., 108°C) was chosen
and stratified by BW into 6 groups of 30 chicks each. because in previous experiments conducted in our de-
Thirty-six uniform groups (6 from each BW block) of partment, the use of higher temperatures to autoclave
6 chicks each were formed and housed in battery cages finely ground PPC did not result in any improvement
(1.0 m × 0.9 m, Avícola Grau, S.A., Madrid, Spain). in growth performance of the broilers. All the 10 sub-
One cage from each of the 6 BW groups was randomly batches corresponding to each of the 2 PPC varieties
assigned to each of the experimental diets. All the cages were thoroughly mixed for uniformity of the composite
were equipped with 2 drinker cups and an open trough sample. The determined chemical analyses of the pro-
feeder. Room temperature was kept at 33°C during the tein sources are shown in Table 1.
first 3 d of life and then was reduced gradually ac- From d 1 to 17, all chicks received a common com-
cording to age until reaching 24°C at 21 d. The chicks mercial diet based on corn and SBM in crumble form,
were kept on a 18 h/d light program and had free ac- and from d 18 to 21 they were fed their respective ex-
cess to feed in mash form and water throughout the perimental diets in mash form on an ad libitum basis.
1850 Frikha et al.
Table 1. Determined chemical composition (% as fed-basis, unless otherwise indicated)1 of fermented soybean meal (FSBM) and pea
protein concentrates (PPC)
PPC-1 PPC-2

Item FSBM Unheated Autoclaved2 Unheated Autoclaved

DM 91.0 92.2 90.7 89.6 90.4

Gross energy (kcal/kg) 4,293 4,202 4,157 4,111 4,179
Total ash 6.7 6.1 6.1 5.6 5.7
Ether extract 0.9 2.5 3.0 2.7 2.4
Neutral detergent fiber 8.5 4.6 3.6 4.1 2.7
Sucrose 0.4 2.9 3.1 3.0 3.2
Raffinose 0.8 0.3 0.7 0.2 0.4
Stachyose 0.9 2.1 1.7 1.9 1.5
Verbascose 0.9 1.4 1.5 1.3 1.4
CP 52.4 45.6 45.5 44.6 45.0
Indispensable amino acid
Arg 3.66 4.12 4.03 4.10 4.13
His 1.36 1.20 1.13 1.12 1.16
Ile 2.32 1.96 1.90 1.86 1.89
Leu 3.99 3.38 3.30 3.22 3.28
Lys 2.69 3.48 3.40 3.28 3.35
Met 0.73 0.45 0.42 0.44 0.40
Phe 2.62 2.30 2.24 2.19 2.16
Thr 2.06 1.77 1.73 1.72 1.69
Val 2.46 2.20 2.16 2.10 2.16
Dispensable amino acid
Ala 2.40 2.05 2.00 1.96 1.90
Asp 5.94 5.18 5.12 4.97 5.04
Cys 0.75 0.65 0.58 0.63 0.59
Glu 9.41 7.42 7.35 7.21 7.32
Gly 2.23 1.99 1.96 1.91 1.97
Pro 2.63 1.90 1.95 1.88 1.92
Ser 2.64 2.25 2.18 2.09 2.14
Trypsin inhibitor activity (mg/g) 0.2 9.4 2.8 9.1 5.3
Saponins (g/kg) 0.1 1.4 1.1 1.1 0.9
Urease activity (mg N/g per min 30°C) 0.02 0.00 0.00 0.00 0.00
Protein dispersibility index 9.1 87.6 47.2 81.5 53.2
1Analyzed in triplicate, except for the amino acids that were determined in duplicate. The CV of the analyses varied between 3 and 7%, depending
on the variable considered.
2Autoclaved: 108°C for 8 min at 0.6 bars.

These diets were based on corn starch and sucrose, with in the ileal digesta, and Nutrientf and AIAf were the
the ingredient tested being the only source of protein concentrations of the same dietary components in the
(20% CP) of the diet. Acid-washed diatomaceous earth feed, all of them expressed on a DM basis. The SID
(Celite Hispánica, S.A., Alicante, Spain) was added to was calculated as indicated by Adedokun et al. (2009)
all diets at 2%, as an additional source of acid insoluble by correcting the AID for the endogenous losses of CP
ash (AIA). The determined CP and AA content of the and of each particular AA determined in birds fed the
experimental diets are shown in Table 2. N-free diets: SID = AID + (basal endogenous losses/
Nutrientf).
Collection of Ileal Digesta
Laboratory Analyses
At 21 d of age, all the birds were euthanized by as-
phyxiation in CO2 atmosphere, and the terminal ileum, Samples (100 g) of the protein ingredients were
defined as the segment of the gastrointestinal tract ground through a laboratory mill (model Z-I, Retsch,
from 15 to 2 cm anterior to the ileocecal junction of all Stuttgart, Germany) equipped with a 0.5-mm screen
6 birds from each cage, was clamped to avoid contami- and analyzed for moisture by oven drying (method
nation. The ileal digesta was collected by gently flush- 930.15), N by combustion (method 990.03) using Leco
ing the digesta with distilled water into plastic contain- equipment (model FP-528, Leco Corporation, St. Jo-
ers, pooled, frozen at −20°C, freeze-dried, and stored in seph, MI), total ash by muffle furnace (method 942.05),
airtight containers at room temperature until further and ether extract by Soxhlet (method 920.39) as de-
chemical analyses. scribed by AOAC International (2000). The gross ener-
The AID of CP and AA of the diets were calculated gy was determined using an isoperibol bomb calorime-
as indicated by Stein et al. (2001), according to the ter (model 356, Parr Instrument Company, Moline, IL).
following equation: AID = [100 − (Nutrientd/Nutri- Sucrose and oligosaccharides (raffinose, stachyose, and
entf) × (AIAf/AIAd)], where Nutrientd and AIAd were verbascose) were determined by HPLC (model 3110,
the concentrations of dietary components and of AIA Perkin Elmer Inc., Waltham, MA) as indicated by de
 PEA PROTEIN CONCENTRATE IN BROILERS 1851
Table 2. Ingredient composition and chemical analyses (%, as-fed basis, unless otherwise indicated) of the experimental diets
PPC2-1 PPC-2

Item FSBM1 Unheated Autoclaved3 Unheated Autoclaved N-free diet

Ingredient
FSBM 38.2 — — — — —
PPC-1 — 43.8 43.8 — — —
PPC-2 — — — 44.8 44.8 —
Corn starch 25.6 22.8 22.8 22.3 22.3 42.75
Sucrose 25.6 22.8 22.8 22.3 22.3 42.75
Soybean oil 5.0 5.0 5.0 5.0 5.0 5.0
Sodium chloride 0.2 0.2 0.2 0.2 0.2 0.2
Dicalcium phosphate 1.9 1.9 1.9 1.9 1.9 1.9
Calcium carbonate 1.0 1.0 1.0 1.0 1.0 1.0
Potassium carbonate — — — — — 0.9
Cellulose — — — — — 3.0
Celite4 2.0 2.0 2.0 2.0 2.0 2.0
Premix5 0.5 0.5 0.5 0.5 0.5 0.5
Determined analysis6
Gross energy (kcal/kg) 3,884 3,998 3,930 3,952 3,907 3,612
DM 94.9 94.3 93.2 93.0 93.6 94.9
CP 20.1 20.3 20.2 19.8 20.2 0.01
Indispensable amino acid7
Arg 1.40 1.83 1.79 1.82 1.85 —
His 0.52 0.53 0.50 0.50 0.52 —
Ile 0.89 0.87 0.84 0.83 0.85 —
Leu 1.53 1.50 1.47 1.43 1.47 —
Lys 1.03 1.55 1.51 1.46 1.50 —
Met 0.28 0.20 0.19 0.20 0.19 —
Phe 1.01 1.02 0.99 0.97 0.97 —
Thr 0.79 0.79 0.77 0.76 0.76 —
Val 0.94 0.98 0.96 0.93 0.97 —
Dispensable amino acid7
Ala 0.92 0.91 0.89 0.87 0.85 —
Asp 2.28 2.31 2.27 2.21 2.26 —
Cys 0.29 0.29 0.26 0.28 0.26 —
Glu 3.61 3.30 3.26 3.20 3.28 —
Gly 0.86 0.89 0.87 0.85 0.88 —
Pro 1.01 0.87 0.84 0.83 0.86 —
Ser 1.01 1.00 0.97 0.93 0.96 —
1Fermented soybean meal.
2Pea protein concentrate.
3Autoclaved: 108°C for 8 min at 0.6 bars.
4Acid-washed diatomaceous earth (Celite Hispánica, S.A., Alicante, Spain).
5Provided the following per kilogram of diet: vitamin A (trans-retinyl acetate), 10,000 IU; vitamin D (cholecalciferol), 2,000 IU; vitamin E (all-
3
rac-tocopherol acetate), 20 IU; vitamin K (bisulfate menadione complex), 3 mg; riboflavin, 5 mg; pantothenic acid (d-calcium pantothenate), 10 mg;
nicotinic acid, 30 mg; pyridoxine (pyridoxine·HCl), 3 mg; thiamine (thiamine mononitrate), 1 mg; vitamin B12 (cianocobalamin), 12 μg; d-biotin, 0.15
mg; choline (choline chloride), 300 mg; folic acid, 0.5 mg; Se (Na2SeO3), 0.1 mg; I (KI), 2.0 mg; Cu (CuSO4·5H2O), 10 mg; Fe (FeSO4·7H2O), 30 mg;
Mn (MnSO4·H2O), 100 mg; Zn (ZnO), 100 mg; ethoxyquin, 110 mg.
6Analyzed in triplicate.
7The amino acid contents of the diets were estimated by multiplying the amino acid content of the protein source tested by the determined CP of
the corresponding diet.

Coca-Sinova et al. (2008). The AA contents of the The moisture, gross energy, and N contents of the diets
protein sources were determined by chromatography were measured as indicated previously for the protein
by Evonik-Degussa (Hanau, Germany) using method sources, and the AIA content as recommended by de
994.12 of AOAC International (1995). For determina- Coca-Sinova et al. (2011). The AA content of the diets
tion of Met and Cys concentration, separate samples was not measured but estimated by multiplying the
were oxidized with performic acid before hydrolysis and determined AA content of the protein sources tested
measured as Met sulfone and cysteic acid, respectively. (FSBM and PPC) by the CP content of the respective
Tryptophan was not determined. The neutral detergent diets. The analyses of DM, ash, AIA, N, and AA from
fiber of the protein sources was determined as described ileal digesta samples were determined according to the
by Van Soest et al. (1991) and expressed on an ash-free procedures described previously for ingredients and
basis. The saponin content was analyzed by reversed- diets. All laboratory analyses were done in triplicate
phase HPLC as described by Price et al. (1986) and the except for ileal digesta measurements, which, because
TIA, urease activity, and protein dispersibility index of the small size of the samples, were determined in
(PDI) as indicated by de Coca-Sinova et al. (2008). duplicate.
1852 Frikha et al.

Statistical Analyses claved PPC than for broilers fed the FSBM with that
of broilers fed the unheated PPC being intermediate
Data were analyzed as a randomized complete design (P < 0.05). For all other indispensable AA, the AID
with 5 treatments. The experimental unit was the cage of FSBM was similar to that of the 2 autoclaved PPC
for all traits and differences among treatments were but higher in general than that of the 2 unheated PPC.
considered significant at P < 0.05. When significant Source of PPC affected the AID of Thr (P < 0.05) and
differences were detected, treatment means were sepa- tended (P = 0.08) to affect that of Lys and Met, but
rated using the Tukey’s test. In addition, data of the no effects were detected for the other indispensable AA.
4 PPC diets were analyzed using the GLM procedure Autoclaving increased (P < 0.001) the AID of all indis-
of SAS (SAS Institute, 1990) and main effects (PPC pensable AA of the PPC (e.g., 92.1 vs. 88.8% for Lys,
variety and heat processing) and the interaction were 83.6 vs. 76.5% for Thr, and 85.5 vs. 76.9% for Met) and
studied. Results in tables are reported as least squares of Cys (75.9 vs. 64.7%). An interaction between type
means. and HP of PPC was observed for the AID of CP and
of most of the indispensable AA, including Ile, Met,
RESULTS and Thr (P < 0.05), and Cys (P < 0.01); autoclaving
tended to increase more (P = 0.09) the AID of the AA
Analytical Evaluation of Protein Sources of PPC-1 than that of PPC-2. The SID of CP was low-
and Diets er for broilers fed FSBM or unheated PPC-1 than for
broilers fed unheated or autoclaved PPC-2 (Table 4).
Neutral detergent fiber was higher for the FSBM For Lys, the SID was lower for broilers fed FSBM than
than for the PPC (8.5 vs. 3.8%, as an average), but for broilers fed autoclaved PPC-1 or unheated or auto-
sucrose (0.4 vs. 3.1%, as an average) and stachyose (0.9 claved PPC-2, with broilers fed unheated PPC-1 being
vs. 1.8%, as an average) contents were lower. Auto- intermediate. Type of PPC affected (P < 0.05) the SID
claving decreased the concentration of stachyose of the of Thr and Val and tended to affect that of Lys (P =
PPC (2.0 vs. 1.6%, as an average) but increased that 0.06) and Met (P = 0.08) with higher values observed
of sucrose, raffinose, and verbascose. The FSBM had for PPC-2. Autoclaving increased (P < 0.001) the SID
more CP and AA content than the PPC, except for of all indispensable and dispensable AA of the PPC.
Arg and Lys, which were lower. Autoclaving did not
modify substantially the AA concentration or the AA DISCUSSION
profile of the PPC. The TIA values were higher for the
2 unheated PPC than for the FSBM, with values for The determined chemical composition of the protein
the 2 autoclaved PPC being intermediate (9.3, 0.2, and sources was similar to expected values (Fundación Es-
4.5 mg of TIA/g, respectively). Autoclaving reduced at pañola Desarrollo Nutrición Animal, 2010), except for
different extent the TIA content of the 2 PPC tested Lys of the FSBM that was lower (3.20 vs. 2.69%). Con-
(9.4 vs. 2.8 mg of TIA/g for PPC-1 and 9.1 vs. 5.3 mg sequently, Lys content was unexpectedly lower for the
of TIA/g for PPC-2). Saponin content was lower for FSBM diet than for the PPC diets. The data suggest
the FSBM than for the 2 unheated PPC samples (0.1 that the fermentation process used in the current ex-
vs. 1.4 and 1.1 g/kg, respectively). Autoclaving reduced periment reduced the Lys content of the original SBM,
slightly the saponin content of the PPC (1.4 to 1.1 and a finding that should be considered when evaluating
1.1 to 0.9 g/kg for PPC-1 and PPC-2, respectively). the nutritive value of commercially available FSBM
The PDI values were lower for the FSBM than for the products. On the other hand, all the diets based on
unheated PPC-1 and PPC-2 (9.1, 87.6, and 81.5%). PPC, had similar AA contents reflecting the similar
Autoclaving reduced the PDI values from 87.6 to 47.2% AA profile of the 2 peas varieties used and that all diets
for PPC-1 and from 81.5 to 53.2% for PPC-2. were formulated to have a similar CP content. Auto-
The determined proximal analyses of the experimen- claving reduced the content of stachyose of the PPC
tal diets were close to expected values, confirming that but increased that of sucrose, raffinose, and verbas-
the diets were mixed correctly (Table 2). The AA pro- cose. These results were unexpected because sugars are
files were similar for the 2 PPC varieties. The Lys con- known to be heat-stable (Pedrosa et al., 2012). Howev-
tent was lower for the FSBM diet (1.03%) than for the er, Alonso et al. (2000) and Ramadan (2012) reported
PPC diets (1.48 to 1.53%). In contrast, Met content also that HP affected the concentration of sucrose and
was higher for the FSBM diet than for the PPC diets oligosaccharides in peas and SBM, respectively.
(0.28 vs. 0.19%, respectively). From d 18 to 21, feed intake was lower for broilers fed
unheated PPC than for broilers fed autoclaved PPC,
ID of CP and AA with broilers fed FSBM being intermediate (42.9 vs.
52.2 vs. 49.3 g/d; data not shown). Probably, the bitter
The AID of CP was similar (P > 0.05) for the FSBM taste of the unheated PPC caused an initial rejection
than for the 2 unheated PPC but lower for both than of the feed by the birds, as has been reported in pigs
for the average of the 2 autoclaved PPC (Table 3). by Valencia et al. (2008). Also, the lower than expected
For Lys, AID was higher for broilers fed the auto- Lys content reported for the FSBM might have resulted
Table 3. Apparent ileal digestibility of CP and amino acids (AA) of broilers at 21 d of age
Indispensable AA Dispensable AA

Item CP Arg His Ile Leu Lys Met Phe Thr Val Ala Asp Cys Glu Gly Pro Ser

Diet
FSBM1 83.4yz 92.8x 88.0x 86.9xy 87.8xy 85.7z 88.8x 88.2xy 79.0x 84.8xy 84.9xy 83.7y 65.0z 90.1x 81.8y 85.8xy 85.1x
PPC2-1 unheated 80.1z 89.9y 83.7y 79.5z 80.7z 87.4yz 78.3z 81.2z 73.3y 78.3z 79.3x 83.4y 65.6z 86.9y 78.7y 77.7z 78.6y
PPC-1 autoclaved3 88.1x 94.0x 90.3x 88.9x 89.4x 92.2x 85.9xy 89.5x 83.4x 87.8y 88.6x 89.0x 77.8x 92.2x 87.1x 87.5x 87.2x
PPC-2 unheated 84.3xy 91.8xy 87.2xy 82.7yz 84.0yz 90.2xy 80.5y 83.7yz 79.7x 82.2yz 84.2y 86.9xy 67.8yz 89.9x 83.1y 81.6yz 82.6xy
PPC-2 autoclaved 87.1xy 93.8x 90.0x 87.0xy 88.9xy 92.1x 85.2xy 88.0xy 82.7x 86.7xy 87.8xy 88.8x 74.0xy 92.1x 86.2x 86.0x 85.6x
SEM (n = 6) 0.97 0.61 0.91 1.14 1.06 0.73 1.76 1.23 1.28 1.15 1.04 0.90 1.70 0.63 0.99 1.01 1.19
Main effect
PPC source
  PPC-1 84.1 91.9 87.0 84.2 85.1 89.8 79.6 85.4 78.3y 83.1 84.0 86.2 69.7 89.5y 82.9 82.5 82.9
  PPC-2 85.7 92.8 88.6 84.8 86.1 91.1 82.9 85.8 81.2x 84.2 86.0 87.8 70.9 91.0x 84.6 83.8 84.1
Heat processing
Unheated 82.2y 90.9y 85.4y 81.1y 82.4y 88.8y 76.9y 82.4y 76.5y 80.2y 81.7y 85.1y 64.7y 88.4y 80.9y 79.5y 80.6y
Autoclaving 87.6x 93.9x 90.1x 87.9x 88.8x 92.1x 85.5x 88.8x 83.6x 87.1x 88.2x 88.9x 75.9x 92.1x 86.7x 86.7x 86.4x
SEM (n = 12) 0.69 0.43 0.64 0.81 0.75 0.52 1.24 0.87 0.91 0.81 0.73 0.63 1.20 0.45 0.71 0.71 0.84
Probability
General model *** *** *** *** *** *** *** *** *** *** *** *** *** *** *** *** ***
PPC source NS NS NS NS NS 0.08 0.08 NS * NS 0.07 0.09 NS * NS NS NS
Autoclaving *** *** *** *** *** *** *** *** *** *** *** *** *** *** *** *** ***
PPC × autoclaving * 0.09 0.05 * * 0.05 * NS * * * NS ** * * * *
x–zMeans within a column not sharing a common superscript are significantly different (P < 0.05).
PEA PROTEIN CONCENTRATE IN BROILERS

1Fermented soybean meal.

2Pea protein concentrate.
3Autoclaved: 108°C for 8 min at 0.6 bars.

*P < 0.05; **P < 0.01; ***P < 0.001.

1853
 1854

Table 4. Standardized ileal digestibility of CP and amino acids (AA) of broilers at 21 d of age
Indispensable AA Dispensable AA

Item CP Arg His Ile Leu Lys Met Phe Thr Val Ala Asp Cys Glu Gly Pro Ser

BEL1 4.74 2.47 1.20 2.70 3.62 2.49 0.97 3.45 4.82 3.26 2.82 5.41 1.76 6.83 3.51 2.70 3.84
Diet
FSBM2 91.2y 97.7x 93.5xy 93.5xy 94.5xy 90.5z 96.5x 94.6x 90.2x 92.8xy 92.9xy 89.5y 76.9y 94.5x 89.8y 92.8x 92.7xy
PPC3-1 unheated 88.7y 94.1y 90.0z 87.1y 88.5y 91.8yz 86.0y 88.4y 86.1y 87.1z 88.3z 89.1y 75.0y 92.4z 87.5y 86.6y 87.5z
PPC-1 autoclaved4 95.0x 97.5x 95.5xy 93.9x 95.7x 95.8x 96.2x 94.5x 93.7x 94.2x 95.9x 94.0x 86.6x 96.6x 94.2x 94.9x 93.6xy
PPC-2 unheated 91.6xy 95.2y 92.4yz 89.0y 90.5y 93.9xy 91.0xy 89.6y 90.3x 89.4yz 91.6yz 91.9xy 79.1y 94.4yz 90.3y 89.2y 89.9yz
PPC-2 autoclaved 95.0x 97.6x 95.7x 95.0x 95.2x 96.1x 96.8x 95.4x 94.3x 94.9x 95.9x 94.3x 89.3x 96.9x 94.1x 94.2x 94.3x
SEM (n = 6) 0.81 0.51 0.76 0.95 0.87 0.62 1.52 1.06 0.99 0.94 0.94 0.75 1.38 0.49 0.76 0.78 0.98
Main effect
PPC source
  PPC-1 91.8 95.8 92.7 91.1 92.1 93.8 91.1 91.9 89.9 91.0 92.1 91.7 82.2 94.5y 90.9 90.8 90.9
  PPC-2 93.3 96.4 94.0 91.4 92.8 95.0 93.9 92.0 92.3 91.8 93.8 93.1 82.8 95.7x 92.2 91.7 91.8
Heat-processing
  Unheated 90.1y 94.7y 91.2y 88.0y 89.5y 92.9y 88.5y 89.0y 88.2y 88.3y 90.0y 90.7y 77.0y 93.4y 88.9y 87.9y 88.7y
  Autoclaving 95.0x 97.5x 95.5x 94.4x 95.4x 96.0x 96.5x 94.9x 94.0x 94.5x 95.9x 94.1x 88.0x 96.8x 94.2x 94.6x 94.0x
Frikha et al.

SEM (n = 12) 0.58 0.36 0.54 0.67 0.61 0.44 1.07 0.75 0.71 0.67 0.58 0.53 0.98 0.35 0.54 0.55 0.70
Probability
General model *** *** *** *** *** *** *** *** *** *** *** *** *** *** *** *** ***
PPC source 0.09 NS NS NS NS 0.06 0.08 NS * * NS 0.08 NS * 0.09 NS NS
Autoclaving *** *** *** *** *** *** *** *** *** *** *** *** *** *** *** *** ***
PPC × autoclaving 0.08 NS NS NS NS NS NS NS 0.09 NS NS NS * NS 0.07 * NS
x–zMeans within a column not sharing a common superscript are significantly different (P < 0.05).
1BEL = basal endogenous losses. Amino acid composition (% CP on DM basis) of endogenous protein at the terminal ileum of chicks obtained by using an N-free diet.
2Fermented soybean meal.
3Pea protein concentrate.
4Autoclaved: 108°C for 8 min at 0.6 bars.

*P < 0.05; ***P < 0.001.

 PEA PROTEIN CONCENTRATE IN BROILERS 1855
in lower feed intake of the birds fed the diet based on ing conditions improved AA digestibility of the 2 PPC,
this protein source. but the improvements were more noticeable for PPC-
The AID of all AA, except Lys, were higher for broil- 1 than for PPC-2, a finding that was consistent with
ers fed FSBM than for broilers fed the unheated PPC- the higher reduction in TIA with autoclaving of the
1, confirming the results of Valencia et al. (2009) who PPC-1 (from 9.4 to 2.8 mg/g) compared with PPC-
showed in broilers higher ID of CP and some essential 2 (from 9.1 to 5.3 mg/g). This information suggests
AA, including Met, Thr, Val, and Arg for FSBM than that part of the beneficial effects of autoclaving on AA
for unheated PPC, although that of Lys was not affect- digestibility were related to a reduction in TIA, con-
ed. However, little differences between AID of FSBM sistent with data of Conan and Carré (1989) and van
and unheated PPC2 were detected. Carré et al. (1991) der Poel (1990). In addition, HP modifies the physi-
suggested that the strong cellular cohesion existing in cal characteristics of the protein fraction of the peas,
the pea cotyledon of some varieties of unprocessed peas improving the accessibility of the digestive enzymes
was the main factor responsible for the poor digestibil- to the substrate (Owusu-Ansah and McCurdy, 1991),
ity of CP observed. In this respect, Le Gall et al. (2007) which in turn might result in an improvement in nutri-
reported that certain proteins of peas, namely albumin ent digestibility. In this respect, Kakade et al. (1973)
2 and lectins, remained virtually intact after passing reported that up to 60% of the growth depressing effect
the stomach and the small intestine, contributing to the observed when unheated soybeans were included in rat
decrease in digestibility of pea proteins as compared diets could be attributed to the lower digestibility of
with soy proteins. However, opposite to these observa- the undenatured protein. Probably, in addition to TI
tions, in the current experiment the SID of CP was content, other factors present in the seed affected the
higher in birds fed the 2 PPC than in birds fed FSBM. digestibility of the CP fraction of the PPC in broilers.
Probably, fine grinding increased the digestibility of the The PPC used in the current research had a saponin
proteins of the PPC, as has been demonstrated by Va- content of 1.4 and 1.1 g/kg, respectively, values that
lencia et al. (2008) in pigs fed micronized peas. were within the range of 0.8 to 2.5 g/kg reported by
The AID and SID of most AA was higher for the Daveby et al. (1997) for dehulled peas. The saponins
unheated PPC-2 than for the unheated PPC-1. The present in peas and other legumes impart a bitter taste
reasons for the differences in ID between the 2 PPC to the feed (Bishnoi and Khetarpaul, 1994), reducing
are not known, but might be linked to variations in the palatability and feed intake (Castell et al., 1996). The
physical structure and chemical composition, as well authors have not found any report on the potential
as subtle differences in ANF content of the peas (Mar- negative effects of pea saponins on AA digestibility in
iscal-Landín et al., 2002; Kluth et al., 2005). poultry. Oakenfull (1981) and Sparg et al. (2004) indi-
Air classification concentrates the TI (Owusu-Ansah cated that saponins present in dicotyledonous angio-
and McCurdy, 1991), saponins (Price et al., 1985), and sperms (such as peas) had little toxic effects when fed
oligosaccharides (Vose et al., 1976) in the protein frac- to animals, an observation that is supported by the
tion of the peas, which affects voluntary feed intake and lack of negative effects of the saponins contained in the
nutrient digestibility. Le Guen et al. (1995) reported PPC on the ID of AA reported herein. In this respect,
that pigs fed diets with 1.1 g of TI from peas/kg had Daveby et al. (1998) confirmed the existence of 2 dif-
lower ID of CP than pigs fed diets with only 0.4 g of ferent types of saponins in peas: DDMP and B. Both
TI/kg. Grosjean et al. (2000) indicated that differences types are monodesmoside saponins with a sugar chain
in TI content were the main factor contributing to dif- linked at the C-3 position of the aglycon. The DDMP
ferences in ID of AA in pigs fed pea cultivars varying saponin differs from the B saponin in that it contains a
in TI content from 2.1 to 10.6 g/kg. However, Gdala et DDMP (2,3-dihydro-2,5-dihydroxy-6-methyl-4H-pyran-
al. (1991) in pigs and Kluth et al. (2005) in broilers in- 4-one) moiety at the C-22 position, which confer to the
dicated that the wide variation in ileal N retention and saponin a bitter taste. Although saponins are resistant
AA digestibility observed in animals fed different pea to heat, the DDMP saponins are easily converted to
varieties was independent of their TI content. More- B saponins when high temperatures are applied (Heng
over, Fan et al. (1994) did not find any relation between et al., 2006). Therefore, autoclaving might have con-
TI content of peas and AID of AA in pigs. tributed to the decomposition of most of the DDMP
In the current experiment, mild autoclaving increased saponins present in the unheated PPC into B saponins,
the AID and SID of CP and AA of the PPC, consistent reducing the bitter taste of PPC. The results support
with the changes in the structure and chemical compo- the suggestion that the concentration in TI present in
sition observed with heating, including a reduction in peas is a more important ANF in young chicks than
ANF content (Carré and Conan, 1989; Parera et al., saponin concentration, and thus, the lower AA digest-
2010). Many of the ANF contained in the pea seeds ibility reported for the unheated PPC compared with
are sensitive to heat, and consequently, moderate heat- the autoclaved PPC was probably more related to the
ing is likely to reduce the activity and negative effects higher TI content rather than to the saponin content.
of these compounds on nutrient digestibility (van der In conclusion, CP and AA digestibility of the unheat-
Poel et al., 1992; Rutherfurd and Moughan, 1997). In ed PPC varied according to type of peas considered,
the current research, autoclaving under mild process- being in general higher for unheated PPC-2 than for
1856 Frikha et al.

unheated PPC-1. Also, except for Lys, ID of the AA Fan, M. Z., W. C. Sauer, and S. Jaikaran. 1994. Amino acid and
was higher for the FSBM than for the micronized un- energy digestibility in peas (Pisum sativum) from white-flowered
spring cultivars from growing pigs. J. Sci. Food Agric. 64:249–
heated PPC-1, although few differences between FSBM 256.
and unheated PPC-2 were detected. Autoclaving im- Fundación Española Desarrollo Nutrición Animal. 2010. Normas
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la Fabricación de Piensos Compuestos. 3rd ed. C. De Blas, G.
ally above those observed for FSBM. The increase in ID G. Mateos, P. G. Rebollar, ed. Fund. Esp. Desarr. Nutr. Anim.,
of PPC with autoclaving was probably associated with Madrid, Spain.
a decrease in TIA content, although minor effects of Gatel, F., and F. Grosjean. 1990. Composition and nutritive value
other ANF present in the PPC, as well as modifications of pea for pigs: a review of European results. Livest. Prod. Sci.
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of the structure of the pea proteins with heating, might Gdala, J., L. Buraczewska, and W. Grala. 1991. Factors influencing
also have been contributing factors. ileal and total digestibility of pea protein and amino acids in pigs.
Pages 240–242 in Proc. 6th Int. Symp. Protein Metabolism and
Nutrition, Vol. 2, Herning, Denmark.
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