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By-Products of the Refining of Soybean Oil as Pigment Sources for Poultry
626
F. Q. ENRIQUEZ AND E. ROSS REFERENCES Association of Official Agricultural Chemists, 1960. Official Methods of Analysis. Ninth Edition. Washington, D.C. Duncan, D. B., 1955. Multiple range and multiple F tests. Biometrics 11: 1-42. Seemanthani, K. B., 1962. Two new double-yielding tapiocas. Indian Farm, 12 : 10. Snedecor, G. W., 1956. Statistical Methods, Sth Edition. Iowa State College Press, Ames, Iowa. Squibb, R. L., and M. K. Wyld, 1951. Effect of yuca meal in baby chick rations. Turrialba, 1: 298-299. Tabayoyong, T. T., 1935. The value of cassava refuse meal in the ration for growing chicks. Philip. Agric. 24: 509-518. Torres, A. Di P., 1957-58. A raspa de mandioca como sucedaneo dos farelos de trigo na alimentacao de pintos. Anais da Esc. Sup. Agric. "Luiz de Queroz," 14-15: 143-150. Vogt, H., 1966. The use of tapioca meal in poultry rations. World Poultry Sci. J. 22: 113-125.
By-Products of the Refining of Soybean Oil as Pigment Sources for Poultry 1. PIGMENTATION STUDIES WITH BROILERS1 BlANKA LlPSTEIN, S. BOENSTEIN AND P . BUDOWSKI National and University, Institute of Agriculture, Rehovot, Israel (Received for publication September 19, 1966)
L
ITTLE attention has been paid to fats as sources of xanthophylls for the pigmentation of chicks. Fritz (1962) stated that the role of added fat in this connection was controversial. Stable or stabilized fats tend to protect xanthophylls against oxidation and may improve their absorption. On the other hand, by raising the energy level of rations, fat supplements cause less feed 1 Contribution from The National and University Institute of Agriculture, Rehovot, Israel. 1966 Series, No. 10S8-E. Part of a Ph.D. thesis to be submitted by the senior author to the Hebrew University of Jerusalem, Faculty of Agriculture, Rehovot, Israel.
to be eaten; thus, the birds will also consume less xanthophylls. On the whole, opinions are divided with respect to the effect of fat supplements (mainly tallow) on broiler pigmentation. Some authors (Fritz et al., 1957; Day and Williams, 1958; Elrod et al, 1958) have reported positive results, whereas others (Hammond and Harshaw, 1941; Ratcliff et al, 1959; Carver, 1959; Couch et al., 1963) denied the value of fat supplements in this respect or have obtained negative results. Papers pealing with the feeding of acidulated soapstocks to chicks have concentrated chiefly on the question of energy supply
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nutritive value of cassava root meal in rations containing 5% tuna, 5% meat and bone meal and 23 to 32.8% soybean meal. Poorer growth and feed conversion at 3 weeks of age were observed with increasing concentrations of cassava root meal in the ration. The addition of molasses or soybean oil to a SO percent cassava ration was without beneficial effect indicating that palatability and essential fatty acid deficiency were not responsible for the poor results. Supplementation of the high cassava ration with 0.15 percent of methionine largely overcame the depressing effects indicating that methionine was the major limiting nutrient. When the ration was balanced with respect to protein and methionine, cassava root meal, at a level of SO percent of the. chick ration, satisfactorily replaced corn.
SOYBEAN OIL BY-PRODUCTS AS PIGMENTORS
Xanthophyll retention was determined by recording feed intake over 48 hours and calculating the amount of xanthophylls consumed. Excreta were collected quantitatively during the same 48-hours period and their xanthophyll content determined on the day of collection. Xanthophyll assays were conducted as follows: In feed: (a) saponification with alcoholic potassium hydroxide, followed by extraction with petroleum ether at 60-90°C; (b) chromatography on a magnesium oxide column; (c) removal of carotene from the column with acetone petroleum ether (1:9); (d) elution of xanthophylls with a 1:1 mixture of petroleum ether and ethanol; and (e) measurement of the absorbancy of the eluate in a Baush & Lomb "Spectronic" colorimeter at 450 mjA. The readings were compared to a standard curve obtained with pure ^-carotene. Therefore all xanthophyll values reported are ^-carotene equivalents. In excreta: A 50-gram representative sample was blended in a high-speed blender with 50 ml. water, 50 ml. 95% ethanol and exactly 100 ml. petroleum ether (60EXPERIMENTAL AND RESULTS 90°C). The mixture was centrifuged Methods. Pigmentation was evaluated until a clear separation of the petroleum both by xanthophyll assays and by visual ether layer was achieved. A suitable aliquot scoring at 33-40 days of age. At that time of this layer was chromatographed as 6-10 ml. blood was withdrawn by heart above. puncture from chicks of average body For brevity sake this report does not inweight. The blood was centrifuged and the clude data obtained on weights and converplasma was assayed for total carotenoids sions (Lipstein, 1967). These results correby the method described by Moore (1957). spond closely to those published previously Four days later their shank and toe-web (Bornstein and Lipstein, 1961, 1963). pigmentation was determined from disks, 7 Results of all four trials were subjected mm. in diameter, cut from the shank skin to analysis of variance according to Snedeand from the web between the second and cor (1956) and to Duncan's multiple range third toes, respectively, by means of a cork test (1955). All interpretations of data are borer (Day and Williams, 1958). based on the 1% level of probability. Visual color scores of shanks of chicks Trial 1. The aim of the first (prelimiwere estimated independently by the senior nary) experiment was to test the effects of author and a technician, by comparison ASS and commercial (bleached) lecithin on with the color fan of Hoffman-La Roche. the degree of pigmentation of crossbred
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and extent of fatty acid utilization (Curtin and Raper, 1956; Bornstein and Lipstein, 1961, 1963; Pepper et al, 1962; Sibbald et al., 1962a, b; Artman, 1964; Lipstein and Bornstein, 1964). An interesting group of minor constituents found in acidulated soybean soapstock (ASS) is represented by carotenoid pigments, some of which are responsible for yellow pigmentation of chicks. The xanthophyll content of ASS is slightly more than half that of good grade dehydrated alfalfa meal (Budowski, 1961). Bornstein and Lipstein (1963) reported results indicating that ASS might constitute an interesting source of pigments for broilers. Soybean lecithin is another oily by-product of the edible oil-refining industry, being rich in lutein in its unbleached state (Scholfield and Dutton, 1954). The purpose of the present study was to obtain additional data on the influence of ASS and lecithin on pigmentation of broilers. In this connection it also seemed of interest to compare different criteria of broiler pigmentation.
627
628
B. LIPSTEIN, S. BORNSTEIN AND P. BUDOWSKI TABLE 1.—Composition of diets (in %) used in trials 1-3 Trial 1
Ingredients Fat supplements1 Alfalfa meal (dehydr.) Starch (potato flour) Soybean meal (45% protein) Ground sorghum (milo) Wheat bran DL-Methionine Constant ingredients2
A
Trial 2
B 3.0
16.0
—
2.5
2.5
2.50
33.0
29.5
42.0
26.5
29.5
34.0
27.0
29.5
32.0
35.5
20.5
31.5
37.0
41.8
58.3
32.7
64.3
58.3
48.8
63.3
57.8
52.3
44.8
63.8
52.8
41.3
2.5
A
Trial 3
C
B
A
c
3.0
8.0
2.50
2.50
_ —
B
c
D
E
F
G
3.0
6.0
10.0
_
6.0
12.0
3.0
3.0
—
—
— -
—•
16.0
3.0
3.0
3.0
3.0
0.05
0.08
0.15
0.05
0.08
0.10
0.06
0.07
0.09
0.10
— 0.07
6.65
6.65
6.65
6.65
6.65
6.65
6.65
6.65
6.65
6.65
6.65
0.10
6.65
6.65
22.4
25.9
21.6
22.5
23.8
21.5
22.2
22.9
23.9
21.4
22.8
24.3
208.9
239.3
203.8
211.3
223.2
198.6
205.7
212.8
221.5
197.6
211.8
226.4
9.3
9.2
9.4
9.4
9.4
9.2
9.3
9.3
9.3
9.3
9.3
9.3
1 2
Variable with regard to source of fat (see text). Fish meal, 3%; ground oyster shells, 1.5%; D.C.P., 1.5%; mineral mix., 0.30%; vitamin mix., 0.25%; coccidiostat, 0.1%. The composition of the mineral and vitamin mixes and of the coccidiostat was given in a previous report (Bornstein and Lipstein, 1963).
male chicks (Cornish X White Rock), fed these supplements beginning with one day of age. Both supplements were included in chick starter diets (Table 1) at two levels (3% and 16%) in a factorial arrangement, together with an unsupplemented control (Table 2). All diets contained alfalfa meal as an additional source of xanthophylls in order to make the diets more "practical." Each one of these S diets was fed to 3 replicates of 24 chicks each, housed in battery-brooders. Six chicks per treatment were chosen for the determination of plas-
ma and toe-web xanthophyll concentrations. The results obtained in this trial are summarized in Table 2 and Fig. 1. ASS produced a marked pigmenting effect which increased significantly with its dietary level. This effect was more conspicuous for plasma xanthophyll levels than for those of shanks or toe-web. Bleached lecithin at the 3% level tended to increase pigmentation slightly (and not significantly), whereas at the 16% level it interfered with pigmentation.
TABLE 2.—The influence of ASS1 and bleached lecithin2 on the concentration of xanthophylls in plasma, toe-web and shanks (trial 1) (Averages of 6 chicks)
Treatments
Control 3 % ASS 16% ASS 3 % Bleached lecithin 16% Bleached lecithin S.E. 1 2 3
Plasma xanthophylls
Assayed xanthophylls in diet (mcg./gm.)
Xanthophyll intake
6.6 12.8 45.0 6.0 6.5
8.4 16.0 51.6 7.3 7.3
152.6°3 267.0'' 814.0" 164.0° 82.3°
84.5" 151.7»b 212.2" 104.0>> 79.1"
45.8"° 98.6" b 107.8" 68.4 ab ° 29.9°
—
—
20.6
27.8
13.9
(mg.)
Toe-web xanthophylls
Shank xanthophylls
(mcg./lOO ml.) (mcg./lOO cm.2) (mcg./lOO cm.2)
Acidulated soybean soapstock, with a xanthophyll content of 260.0 mcg./gm. Devoid of xanthophylls. Any two mean values not having one letter in common differ significantly (P<0.01).
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Calculated _ composition Crude protein 22.5 Prod, energy (Cal./lOO gm.) 198.1 Cal./lOO g. ratio 8.8 % protein
3.0
— 0.09
629
SOYBEAN OIL BY-PRODUCTS AS PIGMENTOES TABLE 3.—The influence of ASS1, bleached lecithin and unbleached lecithin2 on the concentration of xanthophylls in plasma and toe-web {trial 2) {Averages of 4 chicks)
Treatments
Control 3 % ASS 8% ASS 3 % Unbleached lecithin 8% Unbleached lecithin 3 % Bleached lecithin 8% Bleached lecithin
1 2 8
Xanthophyll intake
Plasma xanthophylls
Toe-web xanthophylls
(mg.)
(mcg./lOO ml.)
(mcg./lOO cm.2)
7.8 14.6 25.1 8.2 9.8 7.1 6.9
7.0 14.3 22.8 7.9 8.7 6.4 6.2
220.0" 3 212.0" 359.5' 189.7" 158.6" 199.5" 200.0"
73.1"° 105.6" 146.3' 97.7"° 82.1"° 74.8"° 57.7°
—
—
36.0
6.9
Acidulated soybean soapstock; its xanthophyll content was 244.0 mg./gm. Xanthophyll content, 31.8 mcg./gm. See corresponding footnote to Table 2.
Xanthophyll concentrations in plasma and toe-web increased linearly with total xanthophyll intake (Fig. 1); in the former case the coefficient of correlation was r = 0.999 and in the latter it was r = 0.727 (both significant). Trial 2. This second trial differed from the first one in that it included an additional supplement (unbleached lecithin), and used 8%, rather than 16%, as the second level of supplementation (Table 3). Every diet was fed to 4 replicates of 24 male chicks each (of the above cross), and only one bird per replicate was chosen, according to its body weight, for the determination of plasma and toe-web xanthophyll concentrations. A summary of the results of this trial is presented in Table 3 and Fig. 1. Similar to the previous trial, the effect of ASS levels was again pronounced, however, by Duncan's range test (1955) only the 8% level was significantly different from the basal diet, especially since this time the plasma xanthophyll levels of the control birds were, for an unknown reason, very high. As in trial 1, a high level of dietary bleached lecithin tended to depress slightly the xanthophyll concentration in the toe-web, but unlike the former trial there was no effect
due to 3 % bleached lecithin. The supplementation with unbleached lecithin, at both dietary levels, likewise had no significant effect (presumably due to the low xanthophyll levels involved). In this trial, too, plasma and toe-web xanthophyll concentrations increased linearly with total xanthophyll intake (Fig. 1), but this time, the significant coefficient
Total xnthopfyll inlake (mg)
FIG. 1.—Degrees of pigmentation in broilers due to the feeding of acidulated soapstock (ASS) and lecithin. Upper diagram—trial 1, lower diagram— trial 2. Plasma ( • — • ) and toe-web ( ± A) xanthophyll levels due to dietary ASS; plasma (0 0 0) and toe-web levels (A A A) due to lecithin.
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S.E.
Assayed xanthophylls in diet (mcg./gm.)
630
B . LlPSTEIN, S. BORNSTEIN AND P . BUDOWSKI TABLE 4.-
-The influence of alfalfa meal1, ASS2 and unbleached lecithin* i • the retention of xanthophyll by broiler chicks of two ages {trial 3) 33 days of age
Diet
No.
G
Control 3 % ASS 6% ASS 10% ASS 3 % alfalfa meal 6% A S S + 3 % alfalfa meal 6% A S S + 6 % lecithin
Assayed xanthophylls in diet (mcg./gm.)
Retention 4
(%)
1.3 4.2 10.1 15.9 7.3
50.3 d e 6 59.4" 1 71#7abo
Assayed xanthophylls in diet (mcg./gm.)
Retention 4
(%)
69.2 b °
1.3 5.8 10.4 16.1 6.8
40.4« 74. 7abo 72>b° 66.9 b0 82.8»
17.1
63.8° d
17.8
79.0*b
12.4
61.7 c d
12.8
62.8^
1
Xanthophyll content, 270.0 mcg./gm. Xanthophyll content, 168.6 mcg./gm. Xanthophyll content, 44.6 mcg./gm. * Averages of 4 replicate groups of 10 chicks each. « See footnote 3 to Table 2; S.E. = 2.26. 2
3
of correlation for plasma xanthophylls (r = 0.862) was lower than that for toeweb xanthophyll (r = 0.999). Trial 3. The objective of the third trial was to evaluate the pigmenting effect of ASS, supplied as sole pigmenter at 3 different levels or in combination with alfalfa meal and with unbleached lecithin, as described in Tables 1 and 4. In this trial both male and female crossbred (Cornish X White Rock) chicks were used. One-day-old chicks were allocated at random to the 7 dietary treatments, each diet being fed to 4 replicates of 10 chicks each, 2 replicates containing only male chicks and the other 2 replicates containing only female chicks. In this trial the determinations of xanthophyll concentrations in blood plasma and toe-web were performed at the age of 33 days and were repeated 25 days later. The retention of xanthophylls was determined at the ages of 28-29 and 54-55 days. Table 4 presents a summary of the data obtained on the retention of xanthophylls. There were no differences in the retention between male or female groups within the
same treatment; hence, these data are based on 4 replicates each. At the younger age there appeared to exist a linear relationship between ASS-derived xanthophyll level in the diet and xanthophyll retention, up to a dietary level of 10 mcg./gm. (due to supplementation with 6% ASS), whereas at the older age 4 mcg./gm. dietary xanthophyll already caused maximal retention; the rate of maximal retention at both ages was practically identical. Analysis of variance of the data presented in Table 4 shows that age had a significant effect on the level of xanthophyll retention. This was not true for ASS-derived xanthophylls, but according to the results obtained with diets E and F, xanthophylls derived from alfalfa meal were significantly better retained at 58 days than at 33 days of age. For unexplained reasons there is a discrepancy between the assayed and calculated xanthophyll levels of diet B at the younger age and of diet E at both ages. However, the above effect of age on retention of alfalfaderived xanthophylls is evident irrespective of whether assayed or calculated values are used. A comparison of the xanthophyll reten-
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A B C D E F
Supplements
58 days of age
631
SOYBEAN OIL BY-PRODUCTS AS PIGMENTOES TABLE 5.—The effect of age and sex on the xanthophyll concentration in the plasma and toe-web of broiler chicks at two ages {trial 3)1 (Averages of 5 chicks) Diet No.
Age (days)
A
C
D
E
F
G
Average
1
Toe-web xanthophylls (mcg./lOO cm.2)
Males
Females
Average
Males
Females
Average
33 58
28.8 22.5
21.2 24.4
25.0 23.4
20.8 30.9
32.5 11.0
26.6 20.9
Average
25.6
22.8
24.2
25.8
21.7
23.7
33 58
96.8 88.0
62.0 85.6
79.4 86.8
44.2 55.2
48.8 41.6
46.5 48.4
Average
92.4
73.8
83.1
49.7
45.2
47.5
33 58
136.8 112.4
87.2 112.8
112.0 112.6
67.6 71.5
78.0 68.9
72.8 70.2
Average
124.6
100.0
112.3
69.5
73.5
71.5
33 58
134.4 192.8
118.4 132.0
126.4 162.4
96.2 102.7
91.0 94.9
93.6 98.8
Average
163.6
125.2
144.4
99.5
92.9
96.2
33 58
230.4 235.0
122.4 174.4
176.4 204.7
83.2 104.0
85.8 100.1
84.5 102.1
Average
232.7
148.4
190.5
93.6
92.9
93.3
33 58
234.8 300.8
208.8 265.0
221.8 282.9
128.7 145.6
152.1 151.1
140.4 148.4
Average
267.8
236.9
252.3
137.1
151.6
144.4
33 58
179.0 289.0
141.6 236.0
160.3 262.5
93.6 125.7
101.4 123.5
97.5 124.6
Average
234.0
188.8
211.4
109.6
112.4
111.0
33 58
148.7 177.2
108.8 147.2
128.7 162.2
76.3 90.8
84.2 84.4
80.3 87.6
Average
162.9
128.0
145.4
83.5
84.3
83.9
Nature of treatments and xanthophyll content of ingredients and diets are detailed in Table 4.
tion of diet G with that of diets C or F seems to indicate a non-significant trend for lesser retention of lecithin-derived xanthophylls than ASS-derived xanthophylls by broiler chicks. In Table 5 the data on xanthophyll levels in plasma and toe-web have been arranged according to sex and age, while Fig. 2 presents data pertaining to the 4 diets A through D, arranged by sex. As in the previous trials there was a linear relationship between xanthophyll intake on the one
hand, and xanthophyll levels in plasma or toe-web on the other. When the data for both sexes and both ages are averaged over, and only the results pertaining to diets A through D are included (4 levels of ASS), then the regression equations for plasma and toe-web xanthophylls on dietary xanthophyll levels were, respectively: Y = 35.9 + 9.05 X (r = 0.914) and Y = 24.58 + 6.29 X (r = 0.992), where Y is xanthophyll concentration in plasma or toe-web, respectively, and X is the die-
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B
Plasma xanthophylls (mcg./lOO ml.)
632
B . LlPSTEIN, S. BOENSTEIN AND P . BUDOWSKI
Dietary xanthophylls (>Jg/g)
tary xanthophyll level. Xanthophyll concentrations in the plasma of male birds were significantly higher than in female birds (Table 5). There was also a significant effect due to age, the plasma xanthophyll levels being higher at 58 than at 33 days (Table 5). This effect is absent in diets A and C, slight in diet B and pronounced in diets D, E, F and G. Thus, except for diet G (and possibly diet D ) , the effect of age on plasma xanthophylls is roughly parallel to its effect on xanthophyll retention (Table 4). Toe-web xanthophyll levels were not significantly affected by either sex or age (Table 5). However, if the effect of age is examined only with regard to diets E, F and G, then an unmistakable trend for higher levels at the older age can be noticed, in line with better xanthophyll retention at this age for these diets, with the exception of die G (Table 4). Trial 4. The aim of this trial was to establish the relative pigmentation efficiency of ASS, in comparison with alfalfa meal and yellow corn, by means of the slope analysis (Bartov and Bornstein, 1966). For this purpose each one of these 3 feedstuffs (after prior xanthophylls assay) was added to the diets at levels such as to sup-
At this stage blood and toe-web samples were obtained from 12 chicks per treatment (6 males and 6 females), which had been chosen on the basis of their body weights (which were close to the average group weight of their sex). Visual color score of the shanks of these chicks was also estimated. The results obtained in this trial are summarized in Table 6 and Fig. 3. They demonstrate, as did those of previous trials, that plasma and toe-web xanthophyll concentrations increased in a linear manner with total xanthophyll intake, as did the visual score. As far as both plasma and toe-web xanthophyll concentrations are concerned, there is no significant difference between the 3 dietary xanthophyll supplements, as long as they are used at the lowest level, that is to say, as long as the dietary xanthophyll levels and total xanthophyll intake do not exceed S.2-S.6 mcg./gm. and 8.6 mg., respectively. At the two higher dietary (or intake) levels, the ulitization of ASS-derived xanthophylls is significantly lower than that of alfalfa-or corn-derived xanthophylls, with no significant difference between the latter two xanthophyll supplements. With regard to the visual score, only the highest levels of alfalfa meal and corn used
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FIG. 2.—The effect of ASS-derived xanthophyll levels and sex of chicks on the degree of pigmentation (trial 3.)- Plasma ( • • ) and toe-web (A A) xanthophyll concentrations of males, and plasma (0 0) and toe-web (A — • A) levels of females.
ply three common dietary xanthophyll concentrations (Table 6). ASS was included in the diet at the expense of refined soybean oil, alfalfa meal replaced wheat bran, and yellow corn was used instead of sorghum, in order to minimize variations in the nutritional value of the resulting 9 diets. A diet without any of these xanthophyll supplements served as negative control. Each diet was fed to one group of 24 crossbred (Cornish X White Rock) chicks, comprising equal numbers of males and females, beginning with the first day of age and up to the age of 40 days.
633
SOYBEAN O I L BY-PRODUCTS AS PIGMENTORS TABLE 6.—The influence of ASS1, alfalfa meaP, and yellow corn3 on the concentration of xanthophylls in plasma and toe-web, and on visual score {trial 4) (Average of 12 chicks) Diets 4 Xanthophyll supplements
Xanthophylls in diet
%
Calculated (meg./ gm-)
5
Assayed (meg./ gm.)
Xanthophyll intake (mg.)
Plasma xanthophylls (meg./ 100 ml.)
Toe-web xanthophylls (meg./ 100 cm.2
Visual score
—
1.3
2.2
21.8 re
25.4^
0.15°
1.9 3.8 5.7
3.9 7.9 11.8
5.2 9.3 13.9
8.6 15.2 22.5
70.6° 107. W°
36.6'« 60.9 d e [ 69.4 cde
0.4 b0 1.2abc 1. 7*bo
1.4 2.7 4.0
3.9 7.9 11.8
5.3 8.2 11.6
8.6 13.5 18.8
93.4" 181.4° 259.6»b
49
Alfalfa meal
76.1" 1 110.2"*
0.6 b ° 1. 7 abc 2>b
Yellow corn
17.0 34.0 51.0
3.9 7.9 11.8
5.6 8.7 12.7
8.5 13.6 20.8
104.0 de 206.6 b ° 282.2»
52.5*>f 92.5 b ° 130.7'
1 l abo 2!5 ab0 3.5"
—
—
—
—
11.40
5.00
ASS
S.E.
1 6 0 . &<*>•
4 ef
0.47
1
Acidulated soybean soapstock; its xanthophyll content was 209.3 meg./gm. Xanthophyll content, 294.0 meg./gm. 8 Xanthophyll content, 23.2 meg./gm. 4 The diets contained 6.0% total oils, 4.0% wheat bran or alfalfa meal (or a combination of both), and 51.0% total grains; constant ingredients were soybean meal (45% protein) 27.5%; fish meal, 9.0%; ground oyster shells, 1.1%; D.C.P., 0.6%; mineral mix., 0.30%; vitamin mix., 0.35%; coccidiostat, 0.10%. 6 Calculated only from the three main xanthophyll sources (ASS, alfalfa meal and yellow corn). Small additional amounts of xanthophylls are present in some of the other ingredients of the basal diet. 6 See footnote 3 to Table 2. 2
in this trial gave significantly better results than the unsupplemented control diet, and only the highest level of corn gave significantly better results than the lowest levels of ASS and alfalfa meal. In spite of this relative lack of statistically significant differences, the visual score exhibits distinct linear relationships with xanthophyll intakes (Table 6 and Fig. 3). Table 7 presents the regression equations and the relative xanthophyll utilization (based on "b" of these equations) when the utilization of corn-derived xanthophylls is considered as 100%. On the basis of these data the relative efficiency (yellow corn = 100%) of ASS and alfalfa meal as xanthophyll supplements ranges from 40 to 46% and from 84 to 100%, respectively, depending on the criterion used for evaluating pigmentation. Plasma xanthophyll levels gave the highest relative ratings, that
is to say, the least differences between the supplements (Fig. 3), while there is little difference between toe-web xanthophyll concentrations or visual score as a basis for evaluating the efficiency of ASS or alfalfa meal relative to yellow corn. In this trial the data for male and female chicks were obtained separately. No differences between the sexes were observed with regard to toe-web xanthophyll concentrations or visual score. However, on the basis of plasma xanthophyll concentrations, the female chicks appeared to utilize cornderived xanthophylls slightly better, whereas the males had a slight advantage with regard to ASS and alfalfa meal, parallel to the corresponding results of the previous trial (Table 5). The results presented in Table 6 are the averages of both sexes, in view of this lack of consistent differences between sexes.
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—
—
634
B . LlPSTEIN, S. BOENSTEIN AND P . BuDOWSKI
FIG. 3.—Plasma and toe-web xanthophyll concentrations and visual scores due to 3 levels each of dietary ASS ( • • ) , alfalfa meal ( • •) and yellow corn (A A ) ; trial 4. DISCUSSION
Most of the pigmenters studied heretofore have been restricted to nonfatty materials. The present study deals with two fatty industrial by-products. The first of these, ASS, contained 168-260 meg. xanthophylls per gram in the present trials, in agreement with the data of Budowski et al. (1964). The composition of ASS xanthophylls is rather peculiar, since some of them are artifacts formed during the industrial acidulation of raw soybean soapstocks (Budowski et al, 1964). The second product studied, soya lecithin, usually appears on the market in a bleached state, in which case it is devoid of carotenoids. The unbleached lecithin and in trials 2 and 3 contained 31.8 and 44.6 meg. xanthophylls per gram, respectively.
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Dietary xanthophylls (jjg/g)
Other soy lecithin products exist which represent purified materials, e.g., acetoneinsoluble phospholipids, which are essentially carotenoid-free. Thus, when studying the effect of soy lecithin on pigmentation, one should differentiate between the xanthophyll-supplying role of unbleached lecithin (Scholfield and Dutton, 1954) and the action of phospholipids on the absorption or stability of xanthophylls provided by other sources. The role of the different xanthophylls' fractions in broiler pigmentation has not yet been elucidated, although lutein had been considered as one of the important pigments for this purpose (Fritz et al, 19S7; Couch et al, 1962). The total xanthophylls of ASS contain about 13.5— 16.0% lutein, while in lecithin lutein makes up 61.6-80.5% of the total xanthophylls (Lipstein et al, 1967). Scholfield and Dutton (1954) reported that the carotenoids of soya lecithin contained 75% lutein. The relatively good pigmentation results obtained with lecithin in trial 3 (diet G of Table 5) as compared to ASS (diets C and D of Table 5), especially at the older age, may be construed as indicating that lutein is more important than other xanthophylls' fractions as far as broiler pigmentation is concerned. ASS demonstrated a marked capability for broiler pigmentation, relative to lecithin, the degree of pigmentation increasing linearly with dietary ASS levels. The results (Figs. 1 and 2) indicate that up to a level of 10% dietary ASS there is no evidence of a diminishing response. In each one of the four trials there was one treatment containing 2.5-3.0% alfalfa meal as the only xanthophyll supplement. If their pigmentation results are compared with the data reported by Day and Williams (1958), Ratcliff et al (1959) and Combs and Nicholson (1963), a great variability becomes apparent. On the other
635
SOYBEAN OIL BY-PRODUCTS AS PIGMENTORS TABLE 7.—Regression equations for plasma xanthophylls, toe-web xanthophylls and visual score on dietary xanthophyll intake {trial 4) Xanthophyll supplements
Correlation coefficient1
Regression equation
Relative xanthophyll utilization (%)
Plasma xanthophylls Y2 = 8 . 6 0 + 6.72X 3 0.844 0.958 Y = --18.14+14.61X 0.911 Y = - - 9.27+14.49X
46.4 100.8 100.0 4
ASS Alfalfa meal Yellow corn
Y= Y= Y=
Toe-web xanthophylls 19.78+ 2.31X 0.817 10.30+ 5.11X 0.944 9 . 7 0 + 5.79X 0.854
39.9 88.3 100.0
ASS Alfalfa meal Yellow corn
Y = - - 0 . 1 2 + 0.08X Y = - - 0 . 3 3 + 0.16X Y = - - 0 . 3 0 + 0.19X
Visual score 0.715 0.853 0.907
42.1 84.2 100.0
1
All coefficients of correlation are significant (P<0.01). Y=plasma xanthophylls, mcg./lOO ml.; toe-web xanthophylls, mcg./lOO cm2; visual score, units Hoffman-La Roche. 3 X = xanthophyll intake. 4 By definition. 2
hand according to the results of trial 4 (Table 7), the relative utilization of xanthophylls from ASS was found to be only 40-46%, and- that of alfalfa-derived xanthophylls 84-100%, of the utilization of corn xanthophylls, as far as broiler pigmentation is concerned. This high utilization of alfalfa xanthophylls relative to corn xanthophylls (84100%) for pigmentation of broilers is decidedly better than the corresponding data of Day and Williams (1958), Ratcliff et al. (1959) and Waldroup et al. (1960). The yellow corn used in trial 4 was commercial U.S.A.—imported grain, and the availability of its xanthophylls might have been lowered due to storage (Bartov and Bomstein, 1967). Lipstein et al. (1967) estimated that, for egg yolk pigmentation, ASS had an absolute xanthophyll utilization of 6% as compared to 20% utilization of corn xanthophylls, or a relative utilization of 30% as compared to corn; this is in fair agreement with the above values of 40-46% for broiler pigmentation. Lutein makes up 55-65% of the corn xanthophylls (Bartov and
Bornstein, 1967) and 13-16% of the ASSderived xanthyphylls (Lipstein et al., 1967). Thus the xanthophylls of ASS contain only about 27% as much lutein as do those of corn. These results may be considered a second indication, that lutein may play a more important role than other xanthophyll fractions in broiler pigmentation. The effect of age on retention of ASS-derived xanthophylls was slight and not significant. On the other hand, age seems to have a significant effect on the retention of alfalfa xanthophylls (Table 4). The relatively similar retention of ASS- and alfalfaderived xanthophylls, in contrast to the marked difference in their respective utilization (Table 7), suggests that the latter is not due to differences in intestinal absorption, but may possibly be the result of relative predominance of various xanthophyll fractions, of different pigmenting capacity. Changes in degree of retention (Table 4) were reflected clearly in the concentration of xanthophylls in blood plasma, but only slightly so in that of toe-web (Table 5). The latter was not significantly affected by either age or sex, whereas the former was.
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ASS Alfalfa meal Yellow corn
636
B . LlPSTEIN, S. BORNSTEIN AND P . BUDOWSKI
phylls of which are better utilized than those of ASS, the latter contributes calories as well as pigments, and may therefore be added freely to broiler diets, providing good pigmentation and economic growth. SUMMARY
By-products of the refining of soybean oil were tested as sources of pigments in practical broiler rations in four trials. Results obtained indicate that acidulated soybean soapstock (ASS), containing 168-260 mcg./gm. xanthophylls, may serve as a pigmenter for broilers. On the other hand, soya lecithin contained only 32-45 meg./ gm. xanthophylls, and hence cannot serve as a pigmenter for chicks, although lutein comprises a much higher proportion of its xanthophylls. The degree of pigmentation due to ASSderived xanthophylls increased linearly with increasing dietary xanthophyll concentration. However, their relative utilization was only 40%-46% in comparison with the pigmentation efficiency of corn xanthophylls. This relatively low utilization of ASS-derived xanthophylls is not due to a low retention, but might possibly be the result of their relatively low content of lutein. The retention of ASS-derived xanthophylls was not affected by the age of the birds, but that from alfalfa meal was. Changes in rate of retention, as well as effects of age and sex, were reflected in the concentration of blood plasma xanthophylls, but not in toe-web xanthophyll levels. Of the criteria used for evaluating the relative efficiency of xanthophyll supplements, visual scoring was the most sensitive, though its treatment differences barely reached statistically significant levels. Plasma xanthophyll levels, on the other hand, constituted a less sensitive criterion, although more statistically significant responses were obtained, with toe-web xan-
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Collins et al. (1955) and Grabowski and Szuman (1963) also found significantly better pigmentation in male birds. According to Fig. 3 the three criteria of broiler pigmentation were linear functions of dietary xanthophyll levels, irrespective of the xanthophyll supplements employed. Plasma xanthophylls' concentration was least affected by differences in the efficiency of these three supplements (see also Table 7). If these differences are not due to variations in retention, as suggested before, the lack of sensitively of this criterion is not unexpected. Visual scoring, on the other hand, appeared to bring out best these differences. In contrast to this advantage of visual scoring, its grades had the lowest coeffcients of correlation with xanthophyll intake (Table 7) and their treatment differences barely reached statistical significance (Table 6). Therefore, from both points of view, toeweb xanthophyll levels seemed the best criterion under the conditions of trial 4. Combs and Nicholson (1963) found xanthophyll content of toe-web to be a less sensitive criterion than visual score or serum xanthophyll levels. In highly pigmented broilers a comparison between visual scoring and colorimetric assay of toe-web carotenoids may possibly be subject to the same considerations as described by Bornstein and Bartov (1966) for yolk pigmentation. The present study demonstrates that ASS, although containing 67-72% free fatty acids (unpublished data) may constitute a good source of xanthophylls. This does not exclude the possibility that an identical concentration and pattern of carotenoid pigments would result in even better pigmentation were it not for its free fatty acids content (Couch et al., 1963). This question is probably related to the oxidative stability of the fats. In contrast to alfalfa meal, the xantho-
SOYBEAN OIL BY-PRODUCTS AS PIGMENTOES
thophylls being the parameter of choice, under these experimental conditions. ACKNOWLEDGMENTS
REFERENCES Artman, N. R., 1964. Interactions of fats and fatty acids as energy sources for the chicks. Poultry Sci. 4 3 : 994-1004. Bartov, I., and S. Bornstein, 1966. Studies on egg yolk pigmentation. 2. Effect of ethoxyquin on xanthophyll utilization. Poultry Sci. 4 5 : 297305. Bartov, I., and S. Bornstein, 1967. Studies on egg yolk pigmentation. 3. The effect of origin and storage conditions of yellow corn on the relative utilization of its xanthophylls. To be published. Bornstien, S., and I. Bartov, 1966. Studies on egg yolk pigmentation. 1. A comparison between visual scoring of yolk color and colorimetric assay of yolk carotenoids. Poultry Sci. 45: 287296. Bornstein, S., and B. Lipstein, 1961. By-products of refining of soybean oil as fat supplements in practical broiler rations. World's Poultry Sci. J. 17: 167-178. Bronstein, S., and B. Lipstein, 1963. Some unusual waste vegetable oils as fat supplements in practical broiler rations. World's Poultry Sci. J. 19: 172-184. Budowski, P., 1961. Acidulated soybean soapstock, chemical composition in relation to poultry nutrition. Seminar of World's Poultry Sci. Association, Israel Branch. Budowski, P., I. Ascarelli, J. Gross, I. Nir and A. Bondi, 1964. Vitamin A activity of acidulated soybean soapstocks in chicks. J. Am. Oil Chem. Soc. 4 1 : 441-445. Carver, D. S., 1959. Variation in the effects of fat supplements on broiler pigmentation, growth feed conversion. Poultry Sci. 38: 71-76.
Collins, W. M., S. C. Thayer and W. C. Skoglund, 1955. Breed and strain differences in shank pigmentation in growing Poultry Sci. 3 4 : 223-228. Combs, G. F., and J. L. Nicholson, 1963. Report of Maryland broiler S-38. Feedstuffs, 35 ( 1 ) : 36-38. Couch, J. R., A. A. Camp, F. M. Farr and C. R. Creger, 1962. Pigmentation of egg yolks and broilers. Proc. Texas Nutrit. Conf. pp. 124128. Couch, J. R., F. M. Farr and A. A. Camp, 1963. Pigmentation of broilers and egg yolks. Proc. Conf. Distillers Feed Res. Council, pp. 31-39. Curtin, L. V., and J. T. Raper, 1956. Feeding value of hydrolyzed vegetable fats in broiler rations. Poultry Sci. 35:273-278. Day, E. J., and W. P. Williams, Jr., 1958. A study of certain factors that influence pigmentation in broilers. Poultry Sci. 37: 1373-1381. Duncan, D. B., 1955. Multiple range and multiple F tests. Biometrics, 1 1 : 1-42. EIrod, R. C , E. S. Robajdek, R. H. Gledhill, W. N. Witz, G. M. Diser and J. W. Hayward, 1958. Pigmentation studies with broilers and laying hens. Feedstuffs, 30(33): 26-32. Fritz, J. C , 1962. Problems concerning pigmentation of poultry and eggs. Proc. Texas Nutrit. Conf. pp. 102-109. Fritz, J. C , F. D. Wharton, Jr. and L. J. Classen, 1957. Influence of feed on broiler pigmentation. Feedstuffs, 29(43): 18-24. Grabowski, T., and J. Szuman, 1963. Broiler production with intensively yellow pigmented skin. Roczn. Nauk. roln. 83-B-2: 227-245. Hammond, J. C , and H. M. Harshaw, 1941. Factors influencing shank and skin color in the growing chicken. Poultry Sci. 20: 437-444. Lipstein, B., 1967. Part of a Ph.D. thesis to be submitted to the Hebrew University of Jerusalem. Lipstein, B., and S. Bornstein, 1964. Studies with acidulated cottonseed-oil soapstock. 1. Its use as a fat supplement in practical broiler rations. Poultry Sci. 43 : 686-693. Lipstein, B., P. Budowski and S. Bornstein, 1967. By-products of the refining of soybean oil as pigment sources for poultry rations. 2. Egg yolk pigmentation. Unpublished data. Moore, T., 1957. Vitamin A. Elsevier Publ. Comp., New York, pp. 586. Pepper, W. F., S. J. Slinger and I. R. Sibbald, 1962. A comparison of feed grade tallow and a mixture of tallow and acidulated soapstocks in practical chicken roaster rations. Poultry Sci. 41:1163-1168.
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The authors are indebted to the "EtzHazait" Oil and Soap Industry, Tel Aviv, for supplying the soybean-oil soapstock and lecithin used in this study. They gratefully acknowledge the technical assistance of Mrs. Miriam Mosheh. This study was supported by a grant from the Committee for the Encouragement of Research and Postgraduate Studies of the General Federation of Jewish Labour.
637
638
B . LlPSTElN, S. BORNSTEIN AND P . BuDOWSKI
Ratcliff, R. G., E. J. Day and J. E. Hill, 1959. Broiler pigmentation as influenced by dietary modifications. Poultry Sci. 38: 1039-1048. Scholfield, C. R., and H. H. Dutton, 1954. Sources of color in soybean lecithin. J. Am. Oil Chem. Soc. 3 1 : 258-260. Sibbald, I. R., W. F. Pepper and S. J. Slinger, 1962a. A comparison of feed grade tallow and acidulated soapstocks in practical chick starter rations. Poultry Sci. 4 1 : 120-124. Sibbald, I. R., S. J. Slinger and G. C. Ashton,
1962b. The utilization of a number of fats, fatty materials and mixtures thereof evaluated in terms of metabolizable energy, chick weight gains and gain: feed ratios. Poultry Sci. 41 : 46-61. Snedecor, G. W., 1956. Statistical Methods, 5th ed. Iowa State College Press, Ames, Iowa. Waldroup, P. W., C. R. Douglas, J. T. McCall and R. H. Harms, 1960. The effect of santoquin on the performance of broilers. Poultry Sci. 39: 1313-1317.
W. M. REID AND R. N. BREWER Department of Poultry Science and The Poultry Disease Research Center, University of Georgia, Athens, Georgia 30601 (Received for publication September 19. 1966)
N
INETEEN or more coccidiostatsi have been manufactured and marketed in the United States to mix in poultry feeds for prevention of coccidiosisi (Reid, 1961). Such materials are so commonly accepted that chick starter feedI minus a coccidiostat is mixed only on spe-• cial order by most commercial feed companies. This new class of additives has intro-duced many new problems for feed manufacturers who have become increasingly' more demanding in specifications for new' coccidiostats (Edgar, 1958). Some of the: different properties listed for an ideal cocci-diostat include good mixing properties,, easy methods of chemical detection, non-toxic properties, low cost and an increasing; demand for efficacy against the less knowni intestinal species of coccidia. Increasing; * Meticlorpindol is the common name for a product marketed under the trademark Coyden by the Dow Chemical Company, Midland, Michigan and under U.S. patent No. 3,206,358. ** Journal Series Paper Number 507. College Experiment Station, University of Georgia. This study was partially supported by NSF Grant GB-1499.
evidence of development in coccidia of resistance to coccidiostats has come both from laboratory studies (Waletzky et al., 1954; Pellerdy, 1962; Vegh, 1962) and from feeding experiments (Cuckler and Malenga, 1955; Joyner, 1957; McLoughlin and Gardiner, 1961, 1962; Gardiner and McLoughlin 1963a, b). Thus, resistance has added another reason to search for new coccidiostats. Lack of resistance is a characteristic to be searched for in any new coccidiostat. Studies herein reported compare the efficacy of meticlorpindol with several widely used commercial coccidiostats in birds inoculated with a mixture of six species of oocysts including Eimeria tenella, E. necatrix, E. acervulina, E. mivati, E. brunetti and E. maxima. MATERIALS AND METHODS
In two different trials White Rock chicks (males in trial 1, mixed sexes in trial 2) were placed in wire-floored, electrically heated battery brooders on an open-formula starter mash (University of Georgia).
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Efficacy Studies on Meticlorpindol* as a Coccidiostat**
F. Q. ENRIQUEZ AND E. ROSS REFERENCES Association of Official Agricultural Chemists, 1960. Official Methods of Analysis. Ninth Edition. Washington, D.C. Duncan, D. B., 1955. Multiple range and multiple F tests. Biometrics 11: 1-42. Seemanthani, K. B., 1962. Two new double-yielding tapiocas. Indian Farm, 12 : 10. Snedecor, G. W., 1956. Statistical Methods, Sth Edition. Iowa State College Press, Ames, Iowa. Squibb, R. L., and M. K. Wyld, 1951. Effect of yuca meal in baby chick rations. Turrialba, 1: 298-299. Tabayoyong, T. T., 1935. The value of cassava refuse meal in the ration for growing chicks. Philip. Agric. 24: 509-518. Torres, A. Di P., 1957-58. A raspa de mandioca como sucedaneo dos farelos de trigo na alimentacao de pintos. Anais da Esc. Sup. Agric. "Luiz de Queroz," 14-15: 143-150. Vogt, H., 1966. The use of tapioca meal in poultry rations. World Poultry Sci. J. 22: 113-125.
By-Products of the Refining of Soybean Oil as Pigment Sources for Poultry 1. PIGMENTATION STUDIES WITH BROILERS1 BlANKA LlPSTEIN, S. BOENSTEIN AND P . BUDOWSKI National and University, Institute of Agriculture, Rehovot, Israel (Received for publication September 19, 1966)
L
ITTLE attention has been paid to fats as sources of xanthophylls for the pigmentation of chicks. Fritz (1962) stated that the role of added fat in this connection was controversial. Stable or stabilized fats tend to protect xanthophylls against oxidation and may improve their absorption. On the other hand, by raising the energy level of rations, fat supplements cause less feed 1 Contribution from The National and University Institute of Agriculture, Rehovot, Israel. 1966 Series, No. 10S8-E. Part of a Ph.D. thesis to be submitted by the senior author to the Hebrew University of Jerusalem, Faculty of Agriculture, Rehovot, Israel.
to be eaten; thus, the birds will also consume less xanthophylls. On the whole, opinions are divided with respect to the effect of fat supplements (mainly tallow) on broiler pigmentation. Some authors (Fritz et al., 1957; Day and Williams, 1958; Elrod et al, 1958) have reported positive results, whereas others (Hammond and Harshaw, 1941; Ratcliff et al, 1959; Carver, 1959; Couch et al., 1963) denied the value of fat supplements in this respect or have obtained negative results. Papers pealing with the feeding of acidulated soapstocks to chicks have concentrated chiefly on the question of energy supply
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nutritive value of cassava root meal in rations containing 5% tuna, 5% meat and bone meal and 23 to 32.8% soybean meal. Poorer growth and feed conversion at 3 weeks of age were observed with increasing concentrations of cassava root meal in the ration. The addition of molasses or soybean oil to a SO percent cassava ration was without beneficial effect indicating that palatability and essential fatty acid deficiency were not responsible for the poor results. Supplementation of the high cassava ration with 0.15 percent of methionine largely overcame the depressing effects indicating that methionine was the major limiting nutrient. When the ration was balanced with respect to protein and methionine, cassava root meal, at a level of SO percent of the. chick ration, satisfactorily replaced corn.
SOYBEAN OIL BY-PRODUCTS AS PIGMENTORS
Xanthophyll retention was determined by recording feed intake over 48 hours and calculating the amount of xanthophylls consumed. Excreta were collected quantitatively during the same 48-hours period and their xanthophyll content determined on the day of collection. Xanthophyll assays were conducted as follows: In feed: (a) saponification with alcoholic potassium hydroxide, followed by extraction with petroleum ether at 60-90°C; (b) chromatography on a magnesium oxide column; (c) removal of carotene from the column with acetone petroleum ether (1:9); (d) elution of xanthophylls with a 1:1 mixture of petroleum ether and ethanol; and (e) measurement of the absorbancy of the eluate in a Baush & Lomb "Spectronic" colorimeter at 450 mjA. The readings were compared to a standard curve obtained with pure ^-carotene. Therefore all xanthophyll values reported are ^-carotene equivalents. In excreta: A 50-gram representative sample was blended in a high-speed blender with 50 ml. water, 50 ml. 95% ethanol and exactly 100 ml. petroleum ether (60EXPERIMENTAL AND RESULTS 90°C). The mixture was centrifuged Methods. Pigmentation was evaluated until a clear separation of the petroleum both by xanthophyll assays and by visual ether layer was achieved. A suitable aliquot scoring at 33-40 days of age. At that time of this layer was chromatographed as 6-10 ml. blood was withdrawn by heart above. puncture from chicks of average body For brevity sake this report does not inweight. The blood was centrifuged and the clude data obtained on weights and converplasma was assayed for total carotenoids sions (Lipstein, 1967). These results correby the method described by Moore (1957). spond closely to those published previously Four days later their shank and toe-web (Bornstein and Lipstein, 1961, 1963). pigmentation was determined from disks, 7 Results of all four trials were subjected mm. in diameter, cut from the shank skin to analysis of variance according to Snedeand from the web between the second and cor (1956) and to Duncan's multiple range third toes, respectively, by means of a cork test (1955). All interpretations of data are borer (Day and Williams, 1958). based on the 1% level of probability. Visual color scores of shanks of chicks Trial 1. The aim of the first (prelimiwere estimated independently by the senior nary) experiment was to test the effects of author and a technician, by comparison ASS and commercial (bleached) lecithin on with the color fan of Hoffman-La Roche. the degree of pigmentation of crossbred
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and extent of fatty acid utilization (Curtin and Raper, 1956; Bornstein and Lipstein, 1961, 1963; Pepper et al, 1962; Sibbald et al., 1962a, b; Artman, 1964; Lipstein and Bornstein, 1964). An interesting group of minor constituents found in acidulated soybean soapstock (ASS) is represented by carotenoid pigments, some of which are responsible for yellow pigmentation of chicks. The xanthophyll content of ASS is slightly more than half that of good grade dehydrated alfalfa meal (Budowski, 1961). Bornstein and Lipstein (1963) reported results indicating that ASS might constitute an interesting source of pigments for broilers. Soybean lecithin is another oily by-product of the edible oil-refining industry, being rich in lutein in its unbleached state (Scholfield and Dutton, 1954). The purpose of the present study was to obtain additional data on the influence of ASS and lecithin on pigmentation of broilers. In this connection it also seemed of interest to compare different criteria of broiler pigmentation.
627
628
B. LIPSTEIN, S. BORNSTEIN AND P. BUDOWSKI TABLE 1.—Composition of diets (in %) used in trials 1-3 Trial 1
Ingredients Fat supplements1 Alfalfa meal (dehydr.) Starch (potato flour) Soybean meal (45% protein) Ground sorghum (milo) Wheat bran DL-Methionine Constant ingredients2
A
Trial 2
B 3.0
16.0
—
2.5
2.5
2.50
33.0
29.5
42.0
26.5
29.5
34.0
27.0
29.5
32.0
35.5
20.5
31.5
37.0
41.8
58.3
32.7
64.3
58.3
48.8
63.3
57.8
52.3
44.8
63.8
52.8
41.3
2.5
A
Trial 3
C
B
A
c
3.0
8.0
2.50
2.50
_ —
B
c
D
E
F
G
3.0
6.0
10.0
_
6.0
12.0
3.0
3.0
—
—
— -
—•
16.0
3.0
3.0
3.0
3.0
0.05
0.08
0.15
0.05
0.08
0.10
0.06
0.07
0.09
0.10
— 0.07
6.65
6.65
6.65
6.65
6.65
6.65
6.65
6.65
6.65
6.65
6.65
0.10
6.65
6.65
22.4
25.9
21.6
22.5
23.8
21.5
22.2
22.9
23.9
21.4
22.8
24.3
208.9
239.3
203.8
211.3
223.2
198.6
205.7
212.8
221.5
197.6
211.8
226.4
9.3
9.2
9.4
9.4
9.4
9.2
9.3
9.3
9.3
9.3
9.3
9.3
1 2
Variable with regard to source of fat (see text). Fish meal, 3%; ground oyster shells, 1.5%; D.C.P., 1.5%; mineral mix., 0.30%; vitamin mix., 0.25%; coccidiostat, 0.1%. The composition of the mineral and vitamin mixes and of the coccidiostat was given in a previous report (Bornstein and Lipstein, 1963).
male chicks (Cornish X White Rock), fed these supplements beginning with one day of age. Both supplements were included in chick starter diets (Table 1) at two levels (3% and 16%) in a factorial arrangement, together with an unsupplemented control (Table 2). All diets contained alfalfa meal as an additional source of xanthophylls in order to make the diets more "practical." Each one of these S diets was fed to 3 replicates of 24 chicks each, housed in battery-brooders. Six chicks per treatment were chosen for the determination of plas-
ma and toe-web xanthophyll concentrations. The results obtained in this trial are summarized in Table 2 and Fig. 1. ASS produced a marked pigmenting effect which increased significantly with its dietary level. This effect was more conspicuous for plasma xanthophyll levels than for those of shanks or toe-web. Bleached lecithin at the 3% level tended to increase pigmentation slightly (and not significantly), whereas at the 16% level it interfered with pigmentation.
TABLE 2.—The influence of ASS1 and bleached lecithin2 on the concentration of xanthophylls in plasma, toe-web and shanks (trial 1) (Averages of 6 chicks)
Treatments
Control 3 % ASS 16% ASS 3 % Bleached lecithin 16% Bleached lecithin S.E. 1 2 3
Plasma xanthophylls
Assayed xanthophylls in diet (mcg./gm.)
Xanthophyll intake
6.6 12.8 45.0 6.0 6.5
8.4 16.0 51.6 7.3 7.3
152.6°3 267.0'' 814.0" 164.0° 82.3°
84.5" 151.7»b 212.2" 104.0>> 79.1"
45.8"° 98.6" b 107.8" 68.4 ab ° 29.9°
—
—
20.6
27.8
13.9
(mg.)
Toe-web xanthophylls
Shank xanthophylls
(mcg./lOO ml.) (mcg./lOO cm.2) (mcg./lOO cm.2)
Acidulated soybean soapstock, with a xanthophyll content of 260.0 mcg./gm. Devoid of xanthophylls. Any two mean values not having one letter in common differ significantly (P<0.01).
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Calculated _ composition Crude protein 22.5 Prod, energy (Cal./lOO gm.) 198.1 Cal./lOO g. ratio 8.8 % protein
3.0
— 0.09
629
SOYBEAN OIL BY-PRODUCTS AS PIGMENTOES TABLE 3.—The influence of ASS1, bleached lecithin and unbleached lecithin2 on the concentration of xanthophylls in plasma and toe-web {trial 2) {Averages of 4 chicks)
Treatments
Control 3 % ASS 8% ASS 3 % Unbleached lecithin 8% Unbleached lecithin 3 % Bleached lecithin 8% Bleached lecithin
1 2 8
Xanthophyll intake
Plasma xanthophylls
Toe-web xanthophylls
(mg.)
(mcg./lOO ml.)
(mcg./lOO cm.2)
7.8 14.6 25.1 8.2 9.8 7.1 6.9
7.0 14.3 22.8 7.9 8.7 6.4 6.2
220.0" 3 212.0" 359.5' 189.7" 158.6" 199.5" 200.0"
73.1"° 105.6" 146.3' 97.7"° 82.1"° 74.8"° 57.7°
—
—
36.0
6.9
Acidulated soybean soapstock; its xanthophyll content was 244.0 mg./gm. Xanthophyll content, 31.8 mcg./gm. See corresponding footnote to Table 2.
Xanthophyll concentrations in plasma and toe-web increased linearly with total xanthophyll intake (Fig. 1); in the former case the coefficient of correlation was r = 0.999 and in the latter it was r = 0.727 (both significant). Trial 2. This second trial differed from the first one in that it included an additional supplement (unbleached lecithin), and used 8%, rather than 16%, as the second level of supplementation (Table 3). Every diet was fed to 4 replicates of 24 male chicks each (of the above cross), and only one bird per replicate was chosen, according to its body weight, for the determination of plasma and toe-web xanthophyll concentrations. A summary of the results of this trial is presented in Table 3 and Fig. 1. Similar to the previous trial, the effect of ASS levels was again pronounced, however, by Duncan's range test (1955) only the 8% level was significantly different from the basal diet, especially since this time the plasma xanthophyll levels of the control birds were, for an unknown reason, very high. As in trial 1, a high level of dietary bleached lecithin tended to depress slightly the xanthophyll concentration in the toe-web, but unlike the former trial there was no effect
due to 3 % bleached lecithin. The supplementation with unbleached lecithin, at both dietary levels, likewise had no significant effect (presumably due to the low xanthophyll levels involved). In this trial, too, plasma and toe-web xanthophyll concentrations increased linearly with total xanthophyll intake (Fig. 1), but this time, the significant coefficient
Total xnthopfyll inlake (mg)
FIG. 1.—Degrees of pigmentation in broilers due to the feeding of acidulated soapstock (ASS) and lecithin. Upper diagram—trial 1, lower diagram— trial 2. Plasma ( • — • ) and toe-web ( ± A) xanthophyll levels due to dietary ASS; plasma (0 0 0) and toe-web levels (A A A) due to lecithin.
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S.E.
Assayed xanthophylls in diet (mcg./gm.)
630
B . LlPSTEIN, S. BORNSTEIN AND P . BUDOWSKI TABLE 4.-
-The influence of alfalfa meal1, ASS2 and unbleached lecithin* i • the retention of xanthophyll by broiler chicks of two ages {trial 3) 33 days of age
Diet
No.
G
Control 3 % ASS 6% ASS 10% ASS 3 % alfalfa meal 6% A S S + 3 % alfalfa meal 6% A S S + 6 % lecithin
Assayed xanthophylls in diet (mcg./gm.)
Retention 4
(%)
1.3 4.2 10.1 15.9 7.3
50.3 d e 6 59.4" 1 71#7abo
Assayed xanthophylls in diet (mcg./gm.)
Retention 4
(%)
69.2 b °
1.3 5.8 10.4 16.1 6.8
40.4« 74. 7abo 72>b° 66.9 b0 82.8»
17.1
63.8° d
17.8
79.0*b
12.4
61.7 c d
12.8
62.8^
1
Xanthophyll content, 270.0 mcg./gm. Xanthophyll content, 168.6 mcg./gm. Xanthophyll content, 44.6 mcg./gm. * Averages of 4 replicate groups of 10 chicks each. « See footnote 3 to Table 2; S.E. = 2.26. 2
3
of correlation for plasma xanthophylls (r = 0.862) was lower than that for toeweb xanthophyll (r = 0.999). Trial 3. The objective of the third trial was to evaluate the pigmenting effect of ASS, supplied as sole pigmenter at 3 different levels or in combination with alfalfa meal and with unbleached lecithin, as described in Tables 1 and 4. In this trial both male and female crossbred (Cornish X White Rock) chicks were used. One-day-old chicks were allocated at random to the 7 dietary treatments, each diet being fed to 4 replicates of 10 chicks each, 2 replicates containing only male chicks and the other 2 replicates containing only female chicks. In this trial the determinations of xanthophyll concentrations in blood plasma and toe-web were performed at the age of 33 days and were repeated 25 days later. The retention of xanthophylls was determined at the ages of 28-29 and 54-55 days. Table 4 presents a summary of the data obtained on the retention of xanthophylls. There were no differences in the retention between male or female groups within the
same treatment; hence, these data are based on 4 replicates each. At the younger age there appeared to exist a linear relationship between ASS-derived xanthophyll level in the diet and xanthophyll retention, up to a dietary level of 10 mcg./gm. (due to supplementation with 6% ASS), whereas at the older age 4 mcg./gm. dietary xanthophyll already caused maximal retention; the rate of maximal retention at both ages was practically identical. Analysis of variance of the data presented in Table 4 shows that age had a significant effect on the level of xanthophyll retention. This was not true for ASS-derived xanthophylls, but according to the results obtained with diets E and F, xanthophylls derived from alfalfa meal were significantly better retained at 58 days than at 33 days of age. For unexplained reasons there is a discrepancy between the assayed and calculated xanthophyll levels of diet B at the younger age and of diet E at both ages. However, the above effect of age on retention of alfalfaderived xanthophylls is evident irrespective of whether assayed or calculated values are used. A comparison of the xanthophyll reten-
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A B C D E F
Supplements
58 days of age
631
SOYBEAN OIL BY-PRODUCTS AS PIGMENTOES TABLE 5.—The effect of age and sex on the xanthophyll concentration in the plasma and toe-web of broiler chicks at two ages {trial 3)1 (Averages of 5 chicks) Diet No.
Age (days)
A
C
D
E
F
G
Average
1
Toe-web xanthophylls (mcg./lOO cm.2)
Males
Females
Average
Males
Females
Average
33 58
28.8 22.5
21.2 24.4
25.0 23.4
20.8 30.9
32.5 11.0
26.6 20.9
Average
25.6
22.8
24.2
25.8
21.7
23.7
33 58
96.8 88.0
62.0 85.6
79.4 86.8
44.2 55.2
48.8 41.6
46.5 48.4
Average
92.4
73.8
83.1
49.7
45.2
47.5
33 58
136.8 112.4
87.2 112.8
112.0 112.6
67.6 71.5
78.0 68.9
72.8 70.2
Average
124.6
100.0
112.3
69.5
73.5
71.5
33 58
134.4 192.8
118.4 132.0
126.4 162.4
96.2 102.7
91.0 94.9
93.6 98.8
Average
163.6
125.2
144.4
99.5
92.9
96.2
33 58
230.4 235.0
122.4 174.4
176.4 204.7
83.2 104.0
85.8 100.1
84.5 102.1
Average
232.7
148.4
190.5
93.6
92.9
93.3
33 58
234.8 300.8
208.8 265.0
221.8 282.9
128.7 145.6
152.1 151.1
140.4 148.4
Average
267.8
236.9
252.3
137.1
151.6
144.4
33 58
179.0 289.0
141.6 236.0
160.3 262.5
93.6 125.7
101.4 123.5
97.5 124.6
Average
234.0
188.8
211.4
109.6
112.4
111.0
33 58
148.7 177.2
108.8 147.2
128.7 162.2
76.3 90.8
84.2 84.4
80.3 87.6
Average
162.9
128.0
145.4
83.5
84.3
83.9
Nature of treatments and xanthophyll content of ingredients and diets are detailed in Table 4.
tion of diet G with that of diets C or F seems to indicate a non-significant trend for lesser retention of lecithin-derived xanthophylls than ASS-derived xanthophylls by broiler chicks. In Table 5 the data on xanthophyll levels in plasma and toe-web have been arranged according to sex and age, while Fig. 2 presents data pertaining to the 4 diets A through D, arranged by sex. As in the previous trials there was a linear relationship between xanthophyll intake on the one
hand, and xanthophyll levels in plasma or toe-web on the other. When the data for both sexes and both ages are averaged over, and only the results pertaining to diets A through D are included (4 levels of ASS), then the regression equations for plasma and toe-web xanthophylls on dietary xanthophyll levels were, respectively: Y = 35.9 + 9.05 X (r = 0.914) and Y = 24.58 + 6.29 X (r = 0.992), where Y is xanthophyll concentration in plasma or toe-web, respectively, and X is the die-
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B
Plasma xanthophylls (mcg./lOO ml.)
632
B . LlPSTEIN, S. BOENSTEIN AND P . BUDOWSKI
Dietary xanthophylls (>Jg/g)
tary xanthophyll level. Xanthophyll concentrations in the plasma of male birds were significantly higher than in female birds (Table 5). There was also a significant effect due to age, the plasma xanthophyll levels being higher at 58 than at 33 days (Table 5). This effect is absent in diets A and C, slight in diet B and pronounced in diets D, E, F and G. Thus, except for diet G (and possibly diet D ) , the effect of age on plasma xanthophylls is roughly parallel to its effect on xanthophyll retention (Table 4). Toe-web xanthophyll levels were not significantly affected by either sex or age (Table 5). However, if the effect of age is examined only with regard to diets E, F and G, then an unmistakable trend for higher levels at the older age can be noticed, in line with better xanthophyll retention at this age for these diets, with the exception of die G (Table 4). Trial 4. The aim of this trial was to establish the relative pigmentation efficiency of ASS, in comparison with alfalfa meal and yellow corn, by means of the slope analysis (Bartov and Bornstein, 1966). For this purpose each one of these 3 feedstuffs (after prior xanthophylls assay) was added to the diets at levels such as to sup-
At this stage blood and toe-web samples were obtained from 12 chicks per treatment (6 males and 6 females), which had been chosen on the basis of their body weights (which were close to the average group weight of their sex). Visual color score of the shanks of these chicks was also estimated. The results obtained in this trial are summarized in Table 6 and Fig. 3. They demonstrate, as did those of previous trials, that plasma and toe-web xanthophyll concentrations increased in a linear manner with total xanthophyll intake, as did the visual score. As far as both plasma and toe-web xanthophyll concentrations are concerned, there is no significant difference between the 3 dietary xanthophyll supplements, as long as they are used at the lowest level, that is to say, as long as the dietary xanthophyll levels and total xanthophyll intake do not exceed S.2-S.6 mcg./gm. and 8.6 mg., respectively. At the two higher dietary (or intake) levels, the ulitization of ASS-derived xanthophylls is significantly lower than that of alfalfa-or corn-derived xanthophylls, with no significant difference between the latter two xanthophyll supplements. With regard to the visual score, only the highest levels of alfalfa meal and corn used
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FIG. 2.—The effect of ASS-derived xanthophyll levels and sex of chicks on the degree of pigmentation (trial 3.)- Plasma ( • • ) and toe-web (A A) xanthophyll concentrations of males, and plasma (0 0) and toe-web (A — • A) levels of females.
ply three common dietary xanthophyll concentrations (Table 6). ASS was included in the diet at the expense of refined soybean oil, alfalfa meal replaced wheat bran, and yellow corn was used instead of sorghum, in order to minimize variations in the nutritional value of the resulting 9 diets. A diet without any of these xanthophyll supplements served as negative control. Each diet was fed to one group of 24 crossbred (Cornish X White Rock) chicks, comprising equal numbers of males and females, beginning with the first day of age and up to the age of 40 days.
633
SOYBEAN O I L BY-PRODUCTS AS PIGMENTORS TABLE 6.—The influence of ASS1, alfalfa meaP, and yellow corn3 on the concentration of xanthophylls in plasma and toe-web, and on visual score {trial 4) (Average of 12 chicks) Diets 4 Xanthophyll supplements
Xanthophylls in diet
%
Calculated (meg./ gm-)
5
Assayed (meg./ gm.)
Xanthophyll intake (mg.)
Plasma xanthophylls (meg./ 100 ml.)
Toe-web xanthophylls (meg./ 100 cm.2
Visual score
—
1.3
2.2
21.8 re
25.4^
0.15°
1.9 3.8 5.7
3.9 7.9 11.8
5.2 9.3 13.9
8.6 15.2 22.5
70.6° 107. W°
36.6'« 60.9 d e [ 69.4 cde
0.4 b0 1.2abc 1. 7*bo
1.4 2.7 4.0
3.9 7.9 11.8
5.3 8.2 11.6
8.6 13.5 18.8
93.4" 181.4° 259.6»b
49
Alfalfa meal
76.1" 1 110.2"*
0.6 b ° 1. 7 abc 2>b
Yellow corn
17.0 34.0 51.0
3.9 7.9 11.8
5.6 8.7 12.7
8.5 13.6 20.8
104.0 de 206.6 b ° 282.2»
52.5*>f 92.5 b ° 130.7'
1 l abo 2!5 ab0 3.5"
—
—
—
—
11.40
5.00
ASS
S.E.
1 6 0 . &<*>•
4 ef
0.47
1
Acidulated soybean soapstock; its xanthophyll content was 209.3 meg./gm. Xanthophyll content, 294.0 meg./gm. 8 Xanthophyll content, 23.2 meg./gm. 4 The diets contained 6.0% total oils, 4.0% wheat bran or alfalfa meal (or a combination of both), and 51.0% total grains; constant ingredients were soybean meal (45% protein) 27.5%; fish meal, 9.0%; ground oyster shells, 1.1%; D.C.P., 0.6%; mineral mix., 0.30%; vitamin mix., 0.35%; coccidiostat, 0.10%. 6 Calculated only from the three main xanthophyll sources (ASS, alfalfa meal and yellow corn). Small additional amounts of xanthophylls are present in some of the other ingredients of the basal diet. 6 See footnote 3 to Table 2. 2
in this trial gave significantly better results than the unsupplemented control diet, and only the highest level of corn gave significantly better results than the lowest levels of ASS and alfalfa meal. In spite of this relative lack of statistically significant differences, the visual score exhibits distinct linear relationships with xanthophyll intakes (Table 6 and Fig. 3). Table 7 presents the regression equations and the relative xanthophyll utilization (based on "b" of these equations) when the utilization of corn-derived xanthophylls is considered as 100%. On the basis of these data the relative efficiency (yellow corn = 100%) of ASS and alfalfa meal as xanthophyll supplements ranges from 40 to 46% and from 84 to 100%, respectively, depending on the criterion used for evaluating pigmentation. Plasma xanthophyll levels gave the highest relative ratings, that
is to say, the least differences between the supplements (Fig. 3), while there is little difference between toe-web xanthophyll concentrations or visual score as a basis for evaluating the efficiency of ASS or alfalfa meal relative to yellow corn. In this trial the data for male and female chicks were obtained separately. No differences between the sexes were observed with regard to toe-web xanthophyll concentrations or visual score. However, on the basis of plasma xanthophyll concentrations, the female chicks appeared to utilize cornderived xanthophylls slightly better, whereas the males had a slight advantage with regard to ASS and alfalfa meal, parallel to the corresponding results of the previous trial (Table 5). The results presented in Table 6 are the averages of both sexes, in view of this lack of consistent differences between sexes.
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—
—
634
B . LlPSTEIN, S. BOENSTEIN AND P . BuDOWSKI
FIG. 3.—Plasma and toe-web xanthophyll concentrations and visual scores due to 3 levels each of dietary ASS ( • • ) , alfalfa meal ( • •) and yellow corn (A A ) ; trial 4. DISCUSSION
Most of the pigmenters studied heretofore have been restricted to nonfatty materials. The present study deals with two fatty industrial by-products. The first of these, ASS, contained 168-260 meg. xanthophylls per gram in the present trials, in agreement with the data of Budowski et al. (1964). The composition of ASS xanthophylls is rather peculiar, since some of them are artifacts formed during the industrial acidulation of raw soybean soapstocks (Budowski et al, 1964). The second product studied, soya lecithin, usually appears on the market in a bleached state, in which case it is devoid of carotenoids. The unbleached lecithin and in trials 2 and 3 contained 31.8 and 44.6 meg. xanthophylls per gram, respectively.
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Dietary xanthophylls (jjg/g)
Other soy lecithin products exist which represent purified materials, e.g., acetoneinsoluble phospholipids, which are essentially carotenoid-free. Thus, when studying the effect of soy lecithin on pigmentation, one should differentiate between the xanthophyll-supplying role of unbleached lecithin (Scholfield and Dutton, 1954) and the action of phospholipids on the absorption or stability of xanthophylls provided by other sources. The role of the different xanthophylls' fractions in broiler pigmentation has not yet been elucidated, although lutein had been considered as one of the important pigments for this purpose (Fritz et al, 19S7; Couch et al, 1962). The total xanthophylls of ASS contain about 13.5— 16.0% lutein, while in lecithin lutein makes up 61.6-80.5% of the total xanthophylls (Lipstein et al, 1967). Scholfield and Dutton (1954) reported that the carotenoids of soya lecithin contained 75% lutein. The relatively good pigmentation results obtained with lecithin in trial 3 (diet G of Table 5) as compared to ASS (diets C and D of Table 5), especially at the older age, may be construed as indicating that lutein is more important than other xanthophylls' fractions as far as broiler pigmentation is concerned. ASS demonstrated a marked capability for broiler pigmentation, relative to lecithin, the degree of pigmentation increasing linearly with dietary ASS levels. The results (Figs. 1 and 2) indicate that up to a level of 10% dietary ASS there is no evidence of a diminishing response. In each one of the four trials there was one treatment containing 2.5-3.0% alfalfa meal as the only xanthophyll supplement. If their pigmentation results are compared with the data reported by Day and Williams (1958), Ratcliff et al (1959) and Combs and Nicholson (1963), a great variability becomes apparent. On the other
635
SOYBEAN OIL BY-PRODUCTS AS PIGMENTORS TABLE 7.—Regression equations for plasma xanthophylls, toe-web xanthophylls and visual score on dietary xanthophyll intake {trial 4) Xanthophyll supplements
Correlation coefficient1
Regression equation
Relative xanthophyll utilization (%)
Plasma xanthophylls Y2 = 8 . 6 0 + 6.72X 3 0.844 0.958 Y = --18.14+14.61X 0.911 Y = - - 9.27+14.49X
46.4 100.8 100.0 4
ASS Alfalfa meal Yellow corn
Y= Y= Y=
Toe-web xanthophylls 19.78+ 2.31X 0.817 10.30+ 5.11X 0.944 9 . 7 0 + 5.79X 0.854
39.9 88.3 100.0
ASS Alfalfa meal Yellow corn
Y = - - 0 . 1 2 + 0.08X Y = - - 0 . 3 3 + 0.16X Y = - - 0 . 3 0 + 0.19X
Visual score 0.715 0.853 0.907
42.1 84.2 100.0
1
All coefficients of correlation are significant (P<0.01). Y=plasma xanthophylls, mcg./lOO ml.; toe-web xanthophylls, mcg./lOO cm2; visual score, units Hoffman-La Roche. 3 X = xanthophyll intake. 4 By definition. 2
hand according to the results of trial 4 (Table 7), the relative utilization of xanthophylls from ASS was found to be only 40-46%, and- that of alfalfa-derived xanthophylls 84-100%, of the utilization of corn xanthophylls, as far as broiler pigmentation is concerned. This high utilization of alfalfa xanthophylls relative to corn xanthophylls (84100%) for pigmentation of broilers is decidedly better than the corresponding data of Day and Williams (1958), Ratcliff et al. (1959) and Waldroup et al. (1960). The yellow corn used in trial 4 was commercial U.S.A.—imported grain, and the availability of its xanthophylls might have been lowered due to storage (Bartov and Bomstein, 1967). Lipstein et al. (1967) estimated that, for egg yolk pigmentation, ASS had an absolute xanthophyll utilization of 6% as compared to 20% utilization of corn xanthophylls, or a relative utilization of 30% as compared to corn; this is in fair agreement with the above values of 40-46% for broiler pigmentation. Lutein makes up 55-65% of the corn xanthophylls (Bartov and
Bornstein, 1967) and 13-16% of the ASSderived xanthyphylls (Lipstein et al., 1967). Thus the xanthophylls of ASS contain only about 27% as much lutein as do those of corn. These results may be considered a second indication, that lutein may play a more important role than other xanthophyll fractions in broiler pigmentation. The effect of age on retention of ASS-derived xanthophylls was slight and not significant. On the other hand, age seems to have a significant effect on the retention of alfalfa xanthophylls (Table 4). The relatively similar retention of ASS- and alfalfaderived xanthophylls, in contrast to the marked difference in their respective utilization (Table 7), suggests that the latter is not due to differences in intestinal absorption, but may possibly be the result of relative predominance of various xanthophyll fractions, of different pigmenting capacity. Changes in degree of retention (Table 4) were reflected clearly in the concentration of xanthophylls in blood plasma, but only slightly so in that of toe-web (Table 5). The latter was not significantly affected by either age or sex, whereas the former was.
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ASS Alfalfa meal Yellow corn
636
B . LlPSTEIN, S. BORNSTEIN AND P . BUDOWSKI
phylls of which are better utilized than those of ASS, the latter contributes calories as well as pigments, and may therefore be added freely to broiler diets, providing good pigmentation and economic growth. SUMMARY
By-products of the refining of soybean oil were tested as sources of pigments in practical broiler rations in four trials. Results obtained indicate that acidulated soybean soapstock (ASS), containing 168-260 mcg./gm. xanthophylls, may serve as a pigmenter for broilers. On the other hand, soya lecithin contained only 32-45 meg./ gm. xanthophylls, and hence cannot serve as a pigmenter for chicks, although lutein comprises a much higher proportion of its xanthophylls. The degree of pigmentation due to ASSderived xanthophylls increased linearly with increasing dietary xanthophyll concentration. However, their relative utilization was only 40%-46% in comparison with the pigmentation efficiency of corn xanthophylls. This relatively low utilization of ASS-derived xanthophylls is not due to a low retention, but might possibly be the result of their relatively low content of lutein. The retention of ASS-derived xanthophylls was not affected by the age of the birds, but that from alfalfa meal was. Changes in rate of retention, as well as effects of age and sex, were reflected in the concentration of blood plasma xanthophylls, but not in toe-web xanthophyll levels. Of the criteria used for evaluating the relative efficiency of xanthophyll supplements, visual scoring was the most sensitive, though its treatment differences barely reached statistically significant levels. Plasma xanthophyll levels, on the other hand, constituted a less sensitive criterion, although more statistically significant responses were obtained, with toe-web xan-
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Collins et al. (1955) and Grabowski and Szuman (1963) also found significantly better pigmentation in male birds. According to Fig. 3 the three criteria of broiler pigmentation were linear functions of dietary xanthophyll levels, irrespective of the xanthophyll supplements employed. Plasma xanthophylls' concentration was least affected by differences in the efficiency of these three supplements (see also Table 7). If these differences are not due to variations in retention, as suggested before, the lack of sensitively of this criterion is not unexpected. Visual scoring, on the other hand, appeared to bring out best these differences. In contrast to this advantage of visual scoring, its grades had the lowest coeffcients of correlation with xanthophyll intake (Table 7) and their treatment differences barely reached statistical significance (Table 6). Therefore, from both points of view, toeweb xanthophyll levels seemed the best criterion under the conditions of trial 4. Combs and Nicholson (1963) found xanthophyll content of toe-web to be a less sensitive criterion than visual score or serum xanthophyll levels. In highly pigmented broilers a comparison between visual scoring and colorimetric assay of toe-web carotenoids may possibly be subject to the same considerations as described by Bornstein and Bartov (1966) for yolk pigmentation. The present study demonstrates that ASS, although containing 67-72% free fatty acids (unpublished data) may constitute a good source of xanthophylls. This does not exclude the possibility that an identical concentration and pattern of carotenoid pigments would result in even better pigmentation were it not for its free fatty acids content (Couch et al., 1963). This question is probably related to the oxidative stability of the fats. In contrast to alfalfa meal, the xantho-
SOYBEAN OIL BY-PRODUCTS AS PIGMENTOES
thophylls being the parameter of choice, under these experimental conditions. ACKNOWLEDGMENTS
REFERENCES Artman, N. R., 1964. Interactions of fats and fatty acids as energy sources for the chicks. Poultry Sci. 4 3 : 994-1004. Bartov, I., and S. Bornstein, 1966. Studies on egg yolk pigmentation. 2. Effect of ethoxyquin on xanthophyll utilization. Poultry Sci. 4 5 : 297305. Bartov, I., and S. Bornstein, 1967. Studies on egg yolk pigmentation. 3. The effect of origin and storage conditions of yellow corn on the relative utilization of its xanthophylls. To be published. Bornstien, S., and I. Bartov, 1966. Studies on egg yolk pigmentation. 1. A comparison between visual scoring of yolk color and colorimetric assay of yolk carotenoids. Poultry Sci. 45: 287296. Bornstein, S., and B. Lipstein, 1961. By-products of refining of soybean oil as fat supplements in practical broiler rations. World's Poultry Sci. J. 17: 167-178. Bronstein, S., and B. Lipstein, 1963. Some unusual waste vegetable oils as fat supplements in practical broiler rations. World's Poultry Sci. J. 19: 172-184. Budowski, P., 1961. Acidulated soybean soapstock, chemical composition in relation to poultry nutrition. Seminar of World's Poultry Sci. Association, Israel Branch. Budowski, P., I. Ascarelli, J. Gross, I. Nir and A. Bondi, 1964. Vitamin A activity of acidulated soybean soapstocks in chicks. J. Am. Oil Chem. Soc. 4 1 : 441-445. Carver, D. S., 1959. Variation in the effects of fat supplements on broiler pigmentation, growth feed conversion. Poultry Sci. 38: 71-76.
Collins, W. M., S. C. Thayer and W. C. Skoglund, 1955. Breed and strain differences in shank pigmentation in growing Poultry Sci. 3 4 : 223-228. Combs, G. F., and J. L. Nicholson, 1963. Report of Maryland broiler S-38. Feedstuffs, 35 ( 1 ) : 36-38. Couch, J. R., A. A. Camp, F. M. Farr and C. R. Creger, 1962. Pigmentation of egg yolks and broilers. Proc. Texas Nutrit. Conf. pp. 124128. Couch, J. R., F. M. Farr and A. A. Camp, 1963. Pigmentation of broilers and egg yolks. Proc. Conf. Distillers Feed Res. Council, pp. 31-39. Curtin, L. V., and J. T. Raper, 1956. Feeding value of hydrolyzed vegetable fats in broiler rations. Poultry Sci. 35:273-278. Day, E. J., and W. P. Williams, Jr., 1958. A study of certain factors that influence pigmentation in broilers. Poultry Sci. 37: 1373-1381. Duncan, D. B., 1955. Multiple range and multiple F tests. Biometrics, 1 1 : 1-42. EIrod, R. C , E. S. Robajdek, R. H. Gledhill, W. N. Witz, G. M. Diser and J. W. Hayward, 1958. Pigmentation studies with broilers and laying hens. Feedstuffs, 30(33): 26-32. Fritz, J. C , 1962. Problems concerning pigmentation of poultry and eggs. Proc. Texas Nutrit. Conf. pp. 102-109. Fritz, J. C , F. D. Wharton, Jr. and L. J. Classen, 1957. Influence of feed on broiler pigmentation. Feedstuffs, 29(43): 18-24. Grabowski, T., and J. Szuman, 1963. Broiler production with intensively yellow pigmented skin. Roczn. Nauk. roln. 83-B-2: 227-245. Hammond, J. C , and H. M. Harshaw, 1941. Factors influencing shank and skin color in the growing chicken. Poultry Sci. 20: 437-444. Lipstein, B., 1967. Part of a Ph.D. thesis to be submitted to the Hebrew University of Jerusalem. Lipstein, B., and S. Bornstein, 1964. Studies with acidulated cottonseed-oil soapstock. 1. Its use as a fat supplement in practical broiler rations. Poultry Sci. 43 : 686-693. Lipstein, B., P. Budowski and S. Bornstein, 1967. By-products of the refining of soybean oil as pigment sources for poultry rations. 2. Egg yolk pigmentation. Unpublished data. Moore, T., 1957. Vitamin A. Elsevier Publ. Comp., New York, pp. 586. Pepper, W. F., S. J. Slinger and I. R. Sibbald, 1962. A comparison of feed grade tallow and a mixture of tallow and acidulated soapstocks in practical chicken roaster rations. Poultry Sci. 41:1163-1168.
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The authors are indebted to the "EtzHazait" Oil and Soap Industry, Tel Aviv, for supplying the soybean-oil soapstock and lecithin used in this study. They gratefully acknowledge the technical assistance of Mrs. Miriam Mosheh. This study was supported by a grant from the Committee for the Encouragement of Research and Postgraduate Studies of the General Federation of Jewish Labour.
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Ratcliff, R. G., E. J. Day and J. E. Hill, 1959. Broiler pigmentation as influenced by dietary modifications. Poultry Sci. 38: 1039-1048. Scholfield, C. R., and H. H. Dutton, 1954. Sources of color in soybean lecithin. J. Am. Oil Chem. Soc. 3 1 : 258-260. Sibbald, I. R., W. F. Pepper and S. J. Slinger, 1962a. A comparison of feed grade tallow and acidulated soapstocks in practical chick starter rations. Poultry Sci. 4 1 : 120-124. Sibbald, I. R., S. J. Slinger and G. C. Ashton,
1962b. The utilization of a number of fats, fatty materials and mixtures thereof evaluated in terms of metabolizable energy, chick weight gains and gain: feed ratios. Poultry Sci. 41 : 46-61. Snedecor, G. W., 1956. Statistical Methods, 5th ed. Iowa State College Press, Ames, Iowa. Waldroup, P. W., C. R. Douglas, J. T. McCall and R. H. Harms, 1960. The effect of santoquin on the performance of broilers. Poultry Sci. 39: 1313-1317.
W. M. REID AND R. N. BREWER Department of Poultry Science and The Poultry Disease Research Center, University of Georgia, Athens, Georgia 30601 (Received for publication September 19. 1966)
N
INETEEN or more coccidiostatsi have been manufactured and marketed in the United States to mix in poultry feeds for prevention of coccidiosisi (Reid, 1961). Such materials are so commonly accepted that chick starter feedI minus a coccidiostat is mixed only on spe-• cial order by most commercial feed companies. This new class of additives has intro-duced many new problems for feed manufacturers who have become increasingly' more demanding in specifications for new' coccidiostats (Edgar, 1958). Some of the: different properties listed for an ideal cocci-diostat include good mixing properties,, easy methods of chemical detection, non-toxic properties, low cost and an increasing; demand for efficacy against the less knowni intestinal species of coccidia. Increasing; * Meticlorpindol is the common name for a product marketed under the trademark Coyden by the Dow Chemical Company, Midland, Michigan and under U.S. patent No. 3,206,358. ** Journal Series Paper Number 507. College Experiment Station, University of Georgia. This study was partially supported by NSF Grant GB-1499.
evidence of development in coccidia of resistance to coccidiostats has come both from laboratory studies (Waletzky et al., 1954; Pellerdy, 1962; Vegh, 1962) and from feeding experiments (Cuckler and Malenga, 1955; Joyner, 1957; McLoughlin and Gardiner, 1961, 1962; Gardiner and McLoughlin 1963a, b). Thus, resistance has added another reason to search for new coccidiostats. Lack of resistance is a characteristic to be searched for in any new coccidiostat. Studies herein reported compare the efficacy of meticlorpindol with several widely used commercial coccidiostats in birds inoculated with a mixture of six species of oocysts including Eimeria tenella, E. necatrix, E. acervulina, E. mivati, E. brunetti and E. maxima. MATERIALS AND METHODS
In two different trials White Rock chicks (males in trial 1, mixed sexes in trial 2) were placed in wire-floored, electrically heated battery brooders on an open-formula starter mash (University of Georgia).
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Efficacy Studies on Meticlorpindol* as a Coccidiostat**