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ABSTRACT An experiment was performed to investi- ied from 15.4 (Daka-40) to 28.3 g/kg (Sonac-50). Sub-
gate the effect of animal vs. vegetable protein sources sequently, the effect of a control diet (without PAP)
in the diet of laying hens on the development of hen vs. 4 PAP diets (50 g/kg of CP of animal origin from
performance. A diet containing protein sources of only the same batches as used in the digestibility study) on
vegetable origin was compared with 4 diets, each con- performance was determined. All diets were isocaloric
taining 1 of 4 processed animal proteins (PAP). Two (AMEn = 2,825 kcal/kg) and isonitrogenous (digestible
2608
VEGETABLE VERSUS ANIMAL PROTEIN IN LAYER DIET 2609
2005). The authors argued for the reuse of proteins Experimental Design. A fecal digestibility study was
in nonruminant diets, provided appropriate analytical conducted in laying hens from 24 to 27 wk of age. The
methods are available to distinguish the origin of the experimental period began at 24 wk of age and lasted
different proteins. Because of the species-to-species ban, until the end of wk 27 of age (a period of 4 wk). From
only meat of pork origin may be included in poultry wk 26 onward, the hens received the experimental diets.
diets. In digestibility studies performed earlier, how- In this experiment, fecal digestibility of 4 different pork
ever, the origin of the tested PAP was rarely reported. meat meals was determined: 1) Sonac-50 (Sonac BV,
Therefore, the nutritional value of pork meat meal in a Vion Ingredients Company, Son, the Netherlands),
laying hen diets is unknown. The aim of this study was 2) Sonac-60 (Sonac BV), 3) Daka-40 (Daka Proteins,
to determine the nutritional value of 4 different PAP Ringsted, Denmark), and 4) Daka-58 (Daka Proteins).
sources (experiment 1) and the effects of these PAP These PAP sources varied in chemical composition,
sources on performance (experiment 2) in laying hens. thereby covering the range of commercially available
pork meat meal quality, as shown in Table 1. Crude
MATERIALS AND METHODS protein content, for instance, ranged from 417 to 617 g/
kg, whereas ash content ranged from 183 to 437 g/kg.
Materials and Methods in Experiment 1 In the preexperimental phase (from wk 17 to 23),
(Digestibility Study) a commercial layer meal diet (2,825 kcal/kg, 6.7 g/kg
of digestible lysine) was fed. From wk 24 onward, the
Table 1. Analyzed nutrients of the processed animal proteins used in experiment 1 (g/kg)1
Meat and bone meal Meat meal
Ingredient (g/kg)
Corn 746.8 643.4 646.9 650.0 639.0
Soybean meal, heat-treated 142.6 122.7 123.4 124.0 121.9
Processed animal protein
Sonac-50 186.5
Sonac-60 163.9
Daka-40 220.0
Daka-58 163.5
Limestone 60.8 4.2 25.4 37.3
Oyster shells 30.4 6.6 30.0 30.0 30.0
Monocalcium phosphate 7.2
Salt 4.3 1.3 3.0 1.1 1.9
Premix laying hens2 5.0 5.0 5.0 5.0 5.0
Titanium oxide 1.0 1.0 1.0 1.0 1.0
l-Lysine hydrochloride 0.9
dl-Methionine 0.9
l-Tryptophan 0.1
Analyzed nutrient3 (g/kg)
creta were collected semiquantitatively per cage over a the differences in AMEn and digestibility coefficients
period of 4 × 24 h to determine AMEn and apparent between the basal and PAP-enriched diets.
digestibility of DM, ash, fat, and amino acids. Excreta Analytical Procedures. After defrosting, excreta
samples were frozen and stored at −18°C until chemical samples were homogenized and subsequently, represen-
analysis. The AMEn content was determined by cor- tative samples were taken. All samples were analyzed
recting AME values to zero nitrogen values (Hill and in duplicate. For determination of the DM content in
Anderson, 1958). Therefore, nitrogen retention (nitro- feed and excreta, samples were freeze-dried accord-
gen intake minus nitrogen excretion; De Jonge et al., ing to International Organization for Standardization
2000) was multiplied by the enthalpy of oxidation of (ISO) method number 6496 (International Organiza-
uric acid (8.7 kcal/g) and subtracted from AME. Ap- tion for Standardization, 1998b). After freeze-drying,
parent digestibility of nutrients was calculated using samples were ground to pass a 1-mm screen and kept
the following equation: for analysis. Air-dried samples were dried in a forced-
air oven at 103°C to a constant weight according to
AD (%) = {1 – [(Tifd/Tifc × Nfc/Nfd)]} × 100, [1] ISO method number 6496 (International Organization
for Standardization, 1998b). Kjeldahl nitrogen content
where AD is the apparent digestibility of nutrients, in feed and excreta was measured according to ISO
Tifd and Tifc represent titanium in feed and excreta, method number 5983 (International Organization for
respectively, and Nfd and Nfc are the nutrients in feed Standardization, 1997) in fresh samples. Crude protein
and excreta, respectively. The AMEn and digestibility content was calculated as nitrogen × 6.25. Fecal nitro-
coefficients of the PAP sources were calculated from gen content was determined according to the method
VEGETABLE VERSUS ANIMAL PROTEIN IN LAYER DIET 2611
of De Jonge et al. (2000). Amino acids in feed and 16 h of light and 8 h of dark, whereas the photoperiod
excreta (except tryptophan) were determined after hy- lasted from 1 to 17 h. Health status of the hens was
drolysis with 6 M hydrochloric acid during 23 h. The monitored daily.
hydrolysate was adjusted to pH 2.2. Before hydrolysis, Experimental Design. Pens were allotted to 1 of 5
sulfur-containing amino acids were oxidated overnight dietary treatments, and each treatment had 8 repli-
with performic acid/phenol at 0°C and neutralized with cates, which were equally divided over the 2 rooms. In
sodium disulfite. Amino acids were separated by ion- this study, a control diet without PAP was compared
exchange chromatography and detected photometri- with 4 PAP diets, thereby using the same PAP batches
cally after postcolumn derivatization with ninhydrin by as used in the digestibility experiment: Sonac-50 (treat-
using an automated amino acid analyzer at 570 nm and ment 2), Sonac-60 (treatment 3), Daka-40 (treatment
at 440 nm for proline according to ISO method num- 4), and Daka-58 (treatment 5). The nutritional values
ber 13903 (International Organization for Standard- of these PAP sources (Table 1) that were determined
ization, 2005a). Tryptophan was released by alkaline in the digestibility study were used to optimize the di-
hydrolysis with lithium hydroxide under nitrogen pres- ets for the performance study. All diets were isocaloric
sure at 120°C for 16 h. Tryptophan was separated by (2,825 kcal/kg), had similar digestible essential amino
reversed-phase C18 HPLC with fluorescence detection acid profiles, and met NRC (1994) requirements (Table
with acetate/methanol as eluants and identified and 3). Each PAP-enriched diet contained 50 g/kg of pro-
quantified by UV detection at 280 nm according to ISO tein of animal origin. Because of differences in protein
Housing, Birds, and Management. A total of 480 where Yij is the dependent variable, μ is the overall
non-beak-trimmed 21-wk-old layers (Isa Brown strain) mean, weeki is the fixed effect of week i (i = 21, 22, …
were housed in 2 climate-controlled rooms. Each room 40), treatmentj is the fixed effect of dietary treatment
contained 20 floor pens (0.90 × 1.50 m), and each pen j (j = 1, 2, … 5), and eij is the error term (Genstat,
contained 12 hens. The experimental period was from 2002). The effects of room and pen were added to the
21 to 40 wk of age. The pens were built of wire and random terms of the model. The P-values for week,
hens could see their flock mates in other pens. Each treatment, and week × treatment are presented. Means
pen contained 4 perches, a feeding trough (length 100 were considered significantly different when P < 0.05.
cm), and 3 nipple drinkers. Sand was used as litter on
the floor. A laying nest was placed outside each pen. RESULTS
Throughout the experiment, litter quality was main-
tained by adding new sand monthly. At the beginning Results of Experiment 1
of the experiment, mean BW of the hens was 1,484 ± (Digestibility Study)
52 g (mean ± SD). Feed and water were provided for
ad libitum consumption. Temperature was set at a con- The digestibility coefficients of the 4 PAP sources
stant value of 21°C. Hens received an L:D schedule of showed a large variation (Table 4). Dry matter digest-
2612 van Krimpen et al.
Table 3. Dietary ingredients and analyzed nutrients of the diets used in experiment 2 (g/kg, as fed basis)1
Meat and bone meal Meat meal
Ingredient (g/kg)
Corn 400.0 400.0 400.0 400.0 400.0
Wheat 211.0 159.0 190.5 150.5 192.2
Soybean meal extract (dietary fiber <50) 129.7 48.6 62.2 47.4 66.1
nutrient2
Digestibility coefficient of
DM (%) 38.42 ± 7.76 55.74 ± 4.31 52.63 ± 3.67 64.28 ± 6.26
Organic matter (%) 47.10 ± 8.69 64.63 ± 3.41 53.65 ± 4.99 61.33 ± 6.76
CP (%) 81.43 ± 3.42 90.06 ± 1.93 85.26 ± 1.05 83.37 ± 1.54
Fat (%) 79.65 ± 8.02 81.20 ± 2.32 94.49 ± 3.80 97.20 ± 6.18
Gross energy (%) 55.95 ± 7.40 66.61 ± 2.81 62.01 ± 2.53 65.44 ± 5.58
AMEn (kcal/kg) 1,819 2,566 2,801 3,107
Digestibility coefficient of amino acid2 (%)
Lysine 78.1 ± 3.80 91.5 ± 1.18 81.0 ± 2.52 86.2 ± 3.69
Methionine 75.3 ± 4.23 86.0 ± 1.71 79.9 ± 1.25 82.7 ± 1.81
Cysteine 41.4 ± 12.27 88.4 ± 2.78 64.5 ± 7.29 54.8 ± 6.07
Methionine + cysteine 67.3 ± 5.80 86.7 ± 1.99 76.1 ± 2.62 72.5 ± 3.35
Threonine 68.0 ± 5.29 84.8 ± 2.17 77.8 ± 5.96 79.2 ± 3.92
Tryptophan 67.0 ± 7.43 90.7 ± 2.12 71.5 ± 3.18 72.5 ± 7.58
Isoleucine 67.9 ± 3.26 87.7 ± 3.74 77.1 ± 1.78 76.8 ± 4.24
Arginine 87.2 ± 3.73 94.2 ± 1.41 91.5 ± 0.69 87.5 ± 2.35
Phenylalanine 76.0 ± 6.30 90.8 ± 4.02 80.1 ± 2.92 81.9 ± 2.31
Daka-40 (41%), and Daka-58 (65%), compared with essential amino acids also differed between treatments.
Sonac-50 (88%), was partly caused by high pressure Egg mass production was probably partly affected by
during processing. different intake levels of (non)essential amino acids. Sig-
nificant positive linear regressions were found between
Effect of PAP Sources on Performance intake of digestible aspartic acid (R2 = 0.19), cysteine
(R2 = 0.18), glutamic acid (R2 = 0.16), histidine (R2
Feed intake level of the 2 meat meal diets (Sonac-60 = 0.21), isoleucine (R2 = 0.16), leucine (R2 = 0.12),
and Daka-58) was higher compared with the intake phenylalanine (R2 = 0.13), serine (R2 = 0.15), tyrosine
level of the 2 meat and bone meal diets (Sonac-50 and (R2 = 0.14), valine (R2 = 0.14), and egg mass produc-
Daka-40). Nutritional values of the PAP were deter- tion (g/hen per d). Compared with the control diet, egg
mined in the digestibility study, and these values were weights of the PAP-rich diets were similar (Sonac-60)
used in calculating the diets for the performance study. or (slightly) reduced (Sonac-50, Daka-40, Daka-58).
Therefore, it was assumed that the nutritional values of Several other studies showed reduced egg weights in
the different diets were similar. Hence, these differences PAP-fed hens (Damron et al., 2001; Bozkurt et al.,
in feed intake were not expected because it is generally 2004). This effect might be associated with the amino
known that laying hens adjust their feed intake to their acid concentration of the PAP used (Parsons et al.,
nutritional needs (van Krimpen et al., 2007, 2008, 2009; 1997; Wang and Parsons, 1998b; Shirley and Parsons,
Van der Meulen et al., 2008). It is unknown which spe- 2001). In the current study, however, differences in egg
cific property in the meat and bone meals was respon- weights could only partially be explained by variations
sible for the reduced feed intake levels. in the intake of digestible amino acids. Significant posi-
Laying hen performance differed between treatments; tive linear regressions were found for cysteine (R2 =
performance was most favorable for the Sonac-50 treat- 0.07), glutamic acid (R2 = 0.05), histidine (R2 = 0.04),
ment and most adverse for the Daka-40 treatment. Al- and serine (R2 = 0.05), but levels of declared variance
though contents of the digestible essential amino ac- were very low.
ids were equated for all diets, amino acid intake levels Nutritional factors, such as dietary protein and min-
differed between treatments due to differences in feed eral contents, could affect litter conditions (Veldkamp
intake level. Because of the different feed intake levels et al., 2007; Enting et al., 2009). In our study, we found
and different dietary contents, intake levels of the non- significant linear regressions between litter wetness and
VEGETABLE VERSUS ANIMAL PROTEIN IN LAYER DIET 2615
intake of calcium (R2
= 0.46), chloride (R2
= 0.36), po-
tassium (R2 = 0.56), sodium (R2 = 0.46). Intake of CP
Treatment × week
showed a weak relationship with litter wetness (R2 =
0.321
0.939
<0.001
0.998
0.998
0.758
0.427
0.10). Compared with the hens fed Sonac-60, only the
hens fed meat and bone meal (Daka-40 and Sonac-50)
showed significantly lower intake levels of calcium
(4,730 vs. 4,524 and 4,520 mg/hen per d), potassium
(754 vs. 703 and 705 mg/hen per d), and sodium (174
P-value
and Sonac-60 were provided by Sonac BV (Son, the Netherlands), and Daka-40 and Daka-58 were provided by Daka Proteins (Ringsted, Denmark).
differences, feed intake levels of hens fed the meat and
Treatment
0.005
0.163
0.014
0.001
0.049
0.156
<0.001 bone meal diets were reduced compared with those of
hens fed the meat meal diets, whereas the feed intake
level of hens fed the control diet was intermediate. Dif-
ferences in feed intake and subsequent mineral intake
seemed to be related to differences in litter wetness.
1.509
1.214
0.595
0.754
0.058
48.4ab
124.5a
32.5a
77.6
59.9
acid intake.
Meat meal
ACKNOWLEDGMENTS
2.26ab
Daka-58
122.3ab
47.6bc
32.4a
78.2
58.4
1,781
24.8b
49.2a
118.9c
79.7
59.4
1,795
within a row with different superscripts are significantly different (P < 0.05).
21.1b
119.1c
46.4c
76.5
58.7
1,767
REFERENCES
Berdanier, C. D. 1998. Advanced Nutrition: Micronutrients. CRC
2.17bc
33.0a
1,820
1655.
Bozkurt, M., A. Alcicek, and M. Cabuk. 2004. The effect of dietary
inclusion of meat and bone meal on the performance of laying
hens at old age. S. Afr. J. Anim. Sci. 34:31–36.
CVB. 2005. Feeding Table (Veevoedertabel). Centraal Veevoederbu-
reau, Lelystad, the Netherlands.
Dale, N. 1997. Metabolizable energy of meat and bone meal. J.
Feed conversion ratio (feed/egg; g/g)
1Sonac-50
a–cValues