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Growth Promoting Factors in Malt Distillers Dried Solubles
Growth Promoting Factors in Malt Distillers Dried Solubles 1. THE EFFECT OF THE MINERAL COMPONENTS W. P. J A I T E School of Veterinary Science, University of Bristol AND J. A. WAKELAM The Distillers Company {Biochemicals) Limited, Broadway House, The Broadway, Wimbledon, London, S.W. 19 (Received for publication August 13, 1957)
HE value of corn distillers solubles in the diets of young stock is well established and they are standard ingredients in many American feeds. In addition to the protein, energy and vitamins which they provide, there is strong evidence that they contain also an unidentified growth factor for chicks and poults. This evidence has been the subject of several adequate reviews (Combs et al., 1954; Scott, 1957; Rasmussen et al. 1957; and others) and it will not, therefore, be further reviewed here. In essence, however, the present position seems to be somewhat confused as to the identity or otherwise of the factor(s) provided by fish meals and by-products, milk products, yeast, corn distillers solubles and other fermentation solubles. Several workers have suggested that part, at least, of the growth promoting properties of these various products derives from their mineral components but here again the situation is somewhat obscure. Morrison et al. (1955) and Norris (1955) observed growth responses approximately half as great as those to the intact products when feeding the ash of five supposed growth factor sources. Dannenburg et al. (1955) and Couch (1955) obtained a similar increase when feeding the ash equivalent to 3 percent corn distillers solubles. Briggs (1956)
has criticized the basal diets used in some of these experiments for falling short of the minimum levels laid down by the National Research Council (Bird et al., 1954), and states that he has failed to establish any growth response to the ash of a number of feedstuffs when added to a purified diet. Scott et al. (1955) also failed to secure responses to the ash of corn distillers solubles. Thus it seems that the experiences of different workers with the mineral components of the various supposed growth factor sources has been as confusing as with the intact products themselves. We have discussed this general point in relation to our experience with malt distillers solubles elsewhere (Wakelam and Jaffe, 1958) and are here concerned principally with our work on malt distillers solubles ash. The recovery of malt distillers solubles from the Scottish distilleries is still at an early stage in contrast to the corn distillers solubles which have been recovered ever since the repeal of Prohibition. Accordingly there is little evidence concerning their properties. We have presented certain preliminary findings (Wakelam and Jaffe, 1958; Jaffe and Wakelam, 1956) and in the former have explained the essential differences between the malt and corn distillers' prod-
520
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T
GROWTH FACTORS IN MALT DISTILLERS DRIED SOLUBLES
these several reasons we do not suggest that our results are necessarily relevant to those obtained with corn distillers solubles mentioned above. EXPERIMENTAL METHODS
The basal diet which has been used in this work is shown in Table 1. This diet was designed to support a high level of growth whilst yet being capable of improvement from the addition of distillers solubles—which term, throughout this paper, refers to the recovered solubles from the "pot-stills" of a malt distillery in the Scottish Highlands. The casein and gelatin are standard edible grade material as also is the maize and starch and all
TABLE 1.—Standard diet for growth-factor work on chicks Basal diet Casein Gelatin Maize starch Sucrose Lard Distillers dried grains Mineral mix
Additions 25% 15% 29% 10% 10% 5% 6%
Any additions were made at the expense of maize starch
Mineral CaC0 3 (%) Steamed bone flour (22% Ca 17% P) (%) CaH4-(P04)2H20(%) KC1 (%) NaCl(%) MgS0 4 (%) MnSCv4H20(%) ZnCl2 (g./100-lb.) K I (g./100-lb.) H3BO3 (g./100-lb.) CoS0 4 -7H 2 0 (g./100-lb.) CuSO 4 -5H 2 O(g./100-lb.) FeSO 4 -7H 2 O(g./100-lb.)
Cystine (%) Vitamin A (I.U./lb.) Cholecalciferol (I.U./lb.) a-tocopheryl acetate (mg./lb.) Menaphthone (mg./lb.) Cyanocobalamin (mg./lb.) Thiamine hydrochloride (mg./lb.) Riboflavin (mg./lb.) Calcium pantothenate (mg./lb.) Nicotinic acid (mg./lb.) Choline chloride (mg./lb.) Pyridoxine hydrochloride (mg./lb.) Biotin (mg./lb.) ^>-aminobenzoic acid (mg./lb.) Inositol (mg./lb.) Folic acid (mg./lb.) Procaine benzyl penicillin (mg./lb.) »••• i. • . Mineral mix amount 9.55) 50.00J 6.60\ / 10.42 12.22 6.38 2.78 12 25 6 6 67 880
0.25 4,000 1,000 4.5 2.0 0.008 1.6 2.6 8.4 24.0 1,200.0 2.6 0.08 2.6 450.0 0.5 4.0
Contribution to basal diet when a(Wed at
Ca (%) P(%) Ratio Ca:P K(%) NaCl(%) Mg (mg./lb.) Mn (mg./lb.) Zn (mg./lb.) I (mg./lb.) B (mg./lb.) Co (mg./lb.) Cu (mg./lb.) Fe (mg./lb.)
6 %
1.005 1.005 1.67 0.33 0.73 225 185 3.45 11 0.8 0.8 12 120
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ucts. It should be made clear at this stage that in our studies of the properties of the ashed material we were concerned primarily to exclude the effect of the ash from our work. This, of course, is in contrast to the approach of the workers already cited who were interested in the ash per se. It is our intention to return to the study of the ash at a later stage but at present we use in our basal diet semi-purified ingredients and the birds receive ordinary tap water—no attempt is made to exclude unknown minerals, rather have we sought in formulating our basal diet to include all possible nutrients so long as the addition of malt distillers solubles will still elicit a growth response. For
521
522
W. P. JAFFE AND J. A. WAKELAM
The diets were made up at frequent intervals (usually about one week according to rate of consumption) and were kept in a cold store. These precautions were considered advisable because of the high level of fat included in the basal diet. RESULTS
Experiment 1 was a 4X4 factorial experiment with the following treatments: 1. Control as in Table 1 2. Control+3% spray-dried fish solubles
3. Control+6% malt distillers dried solubles (hereafter referred to as D.D.S.) 4. Control+ash equivalent to 6% D.D.S: Table 2 shows the results in terms of both growth and food conversion efficiency (F.C.E.). The following conclusions may be drawn: a. A good growth response to D.D.S. which is statistically significant (5% level) at 2, 3, 4 weeks. A marked F.C.E. response to D.D.S. which, however, does not reach a significance. b. D.D.S. ash produces a small growth response which at no stage approaches significance—it is at all stages well under half that to D.D.S. The F.C.E. response is even smaller and is in fact negative in the first week. c. The growth response to fish solubles is very small—in most weeks less than to D.D.S. ash. The F.C.E. response is more marked especially at week 4 where it reaches significance. Experiment Z was an 8X2 Randomised Block arrangement with the following treatments: 1. Control as Table 1 but manganese level reduced to 54 mg./lb. 2. Control+ash equivalent to 3 % D.D.S. 3. Control+ash equivalent to 10% D.D.S. 4. Control+1% D.D.S. 5. Control+2% D.D.S. 6. Control+4% D.D.S. 7. Control+6% D.D.S. 8. Control+10% D.D.S. Table 3 shows the results in detail. The mean weights for treatment 2 in brooder B are unusually low and as this was believed to be due to a faulty heater they have been replaced in the analysis by values estimated by minimising the sums of squares for TreatmentsX Brooders. These estimates are shown in brackets in
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these and the dried grains, added to provide roughage, will naturally provide some minerals. It was hoped that by the high level of protein used we would avoid any effect due to the protein content of the solubles. The manganese level shown in Table 1 is that now included as standard—a lower level was used in some of the experiments here reported. The experimental unit comprises two four-tier electrically heated brooders with thermostatic control. Each wire-floored tier is split longitudinally providing sixteen separate compartments. The chicks used were ordinary commercial hatchery stock (Rhode IslandXLight Sussex) which were allocated at random to the sixteen compartments on arrival as day olds. In Experiment 1 of this paper pullets were used, in all other experiments cockerels. All birds were weighed individually on arrival and at weekly intervals for four weeks and both feed and water were available ab libitum. Food consumption was measured on a group basis. No allowance was made for wastage as it had been found that by reducing the amount of food available this could be virtually eliminated. After a few experiments with 4X4 Latin Square layouts it was established that there was no significant effect due to position and subsequent experiments have been of the 8X2 Randomised Block design.
523
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F.C.E.
G R O W T H FACTORS IN M A L T D I S T I L L E R S D R I E D SOLUBLES
t * ** ***
29.6 (33.2)t 40.9 40.0 40.4** 32.8 31.8 32.3 34.3 30.9
68.8
79.5 78.9
79.2
70.8 69.9
70.4
72.8 68.8
39.1
38.6 38.9
38.8
38.0 38.1
38.0
38.5 37.9
A B
A B
A B
A B
A B
A B
36.9 38.3 37.6* 46.4 37.5 42.0*** 44.1 46.9
75.2
84.6 76.1
80.4
82.5 84.5
83.5
37.6
38.2 38.6
38.4
38.4 37.6
38.0 2.7
45.5***
32.6
70.8
73.9 76.4
38.2
37.0 38.1
27.0
151.8 148.6
1.049 1.103
Week 2
1.515** 1.496 1.571
357.5** 371.0 342.7
395.6 409.5 380.6
1.466 1.450 1.504
206.3*
1.534** 1.515 1.560 1.538** 1.531 1.533
356.8** 378.9 387.7 383.3*** 387.2 400.8
395.0
420.8 425.4 439.4
1.430' 1.489 1.421
226.1** 221.4 239.5
263.6 259.6 278.1
0.073
2.5
0.050
8.8
0.059
9.9
406.4***
444.4 1.437* 246.0*** 284.0 1.362* 126.1***
164.1
0.026
1.542**
1.534 1.551 409.6 4C3.2 448.0 440.8 1.402 1.472 255.6 236.5
294.0 274.1 1.371 1.354 127.2 125.0
165.6 162.6
0.987*
1.532** 394.0*** 432.4 1.455* 230.5**
268.8 1.376*
118.0***
156.4
1.477
415.9 425.8
209.0*
1.404 1.456
247.2
222.8 229.3
215.1 202.9
1.532 1.498 362.1 352.9 400.1 391.0
390.2
1.630*
1.629 1.631 348.5 354.4 387.1 393.3
351.4**
2.158** 244.4 (277.0)t*
317.4 249.7 283.6
1.690
1.663 1.716
2.146 (2.169)
312.2
313.4 311.0
278.3 210.6 (275.7)t
353.0
354.3 351.8
F.C.E.
1.522 1.411
1.428*
1.389 1.468
1.895*
1.896 (1.894)
1.634
1.680 1.589
Weight
206.5 206.1
233.0***
234.3 231.8
148.9 (167.l)f
167.4 130.4 (166.8)t
176.7
173.9 179.5
F.C.E.
Week 4 Weight increase
259.8 267.4
253.6 240.8
244.4
244.5 244.2
271.8
272.9 270.7
188.0
206.5 169.5
217.6
214.8 220.3
Weight
Week 3
Weight increase
1.026 0.948
1.355 1.398
1.435*
106.2*** 120.4 115.6
1.400 1.469
1.530
1.483 1.576
1.489
1.528 1.450
1.407*
1.338 1.476
1.726*
1.707 (1.746)
1.566
1.552 1.581
F.C.E.
104.3 108.1
97.1*
100.4 93.8
94.2
94.3 94.0
111.4***
113.2 109.7
75.2 (81.8)t
82.8 67.6 (80.9)t
84.9
84.8 85.0
Weight increase
diet
1.070
158.6 154.2
143.8
1.127 1.000 1.141
141.3 146.2
135.3
138.9 131.7
132.2
132.3 132.1
1.107 1.147
1.312
1.212 1.413
1.246
1.285 1.206
150.2
114.3
1.264
1.076
121.9 106.7
1.241 1.288
125.8
73.1 64.4
39.1 39.1
1.264
34.0 25.3 (32.5)t
67.8
40.9
A B
125.7 125.8
1.238 1.290
27.2 26.7
68.1 67.5
40.9 40.8
Weight
F.C.E.
A B
Weight
Week 1 Weight increase
Initial weight
Means of brooder
Estimated values—see text, Difference from control significant a t 5% probability level. Difference from control significant a t 1 % probability level. Difference from control significant at 0 . 1 % probability level.
Standard error of single pen means
Average
10% D.D.S.
Average
6 % D.D.S.
Average
4 % D.D.S.
Average
2 % D.D.S.
Average
1% D.D.S.
Average
Ash equiv. to 1 0 % D.D.S.
Average
Ash Equiv. to 3 % D.D.S.
Average
Control
Diet
TABLE 3.—Influence of different levels of D.D.S. and D.D.S. ash on growth and food conversion {Experiment 2). Low (Means of 25 chicks per sub-group, weights in grams, food conversion in g.food/g.weight increase)
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GROWTH FACTORS IN MALT DISTILLERS DRIED SOLUBLES
7. Control A + 6 % D.D.S. 8. Control A + 1 0 % D.D.S. Table 4 shows the detailed results from which the following conclusions may be drawn: a. Control A (high manganese) gave considerably greater growth rates and superior F.C.E. than Control B (low manganese). The difference for both growth and conversion were highly significant (0.1% level of probability) at all weeks. b. Both levels of ash appeared to depress growth in this experiment. In the case of the lower level (3% ash equivalent) this depression was statistically significant at 1 and 4 weeks. c. The response to D.D.S. although reasonably consistent approached statistical significance only for the higher levels (6% and 10%) and in the later weeks. At no stage was the response significant at the 5% level of probability. These results were somewhat surprising. Between Experiments 2 and 3 of this paper the brooders had been moved to a new building and this poor response to D.D.S. led us to consider whether the solubles were acting through the gut flora and we were experiencing a "clean/dirty" house effect—comparable to that reported by Coates et al. (1952). This point has been investigated and is reported elsewhere (Wakelam and Taffe, 1958) where we have discussed the matter in some detail and suggested that it may indeed be the case. Since this suggests that Experiment 3 may represent an atypical state of affairs, we deferred confirmation until we felt reasonably certain that the new house had reached a steady state, and then carried out Experiment 4. In Experiment 4 we examined the effect of two levels of D.D.S. ash and a high level of D.D.S. on the two control diets used in Experiment 3. The treatments
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Table 3. The following conclusions may be drawn. a. 10% ash equivalent produces growth responses which are statistically significant ( 1 % level or better) at all weeks. The F.C.E. response is also significant (5% level or better) at all weeks. b. The response to 10% D.D.S. is always considerably greater than that to the equivalent amount of ash and the difference between ash and solubles response is significant (5% level or better) for both growth and conversion at 2 and 4 weeks. c. 3 % ash equivalent shows growth depression and an adverse effect on conversion at all weeks. The effect is significant (5% level) at 4 weeks. d. It is estimated, from the regression equation, that the growth response to 3 % D.D.S. would be considerably greater than that to 3 % ash equivalent at all weeks and that it would be statistically significant (5% level). These results seem to indicate a marked response to a high level of D.D.S. ash and are in many respects more similar to those reported by the various American workers quoted above than to the results of Experiment 1. Experiment 3 was essentially a repeat of Experiment 2 with the manganese level restored to the high value of Table 1 and an additional group (treatment 2) receiving the same diet as the control in Experiment 2. The treatments (8X2 Randomised Block) were: 1. Control A (Manganese content 185 mg./lb.) 2. Control B (Manganese content 54 mg./lb.) 3. Control A+ash equivalent to 3 % D.D.S. 4. Control A+ash equivalent to 10% D.D.S. 5. Control A + 2 % D.D.S. 6. Control A + 4 % D.D.S.
525
A B
A B
A B
A B
A B
40.4
79.4
39.0
3.1
37.8
41.6
43.7 37.1
34.7 41.0
76.0 82.8
41.3 41.8
79.4
36.6
78.2
41.6
82.7 76.2
36.1 37.1
39.0 39.1
35.4
43.0
78.0 78.3
34.9 35.8
78.5 78.1
43.6 42.3
78.3
34.2
75.8
41.6
41.9 41.2
37.2 31.2
78.7 73.0
0.113
1.093
0.992 1.193
1.096
1.108 1.083
1.098
1.121 1.076
1.163
1.174 1.152
1.233
1.187 1.279
1.434
1.530 1.337
2.286***
2.466 2.107
1.080
1.121 1.040
F.C.E.
158.6
165.0 152.2
158.4
154.5 162.3
153.4
154.3 152.5
152.3
152.1 152.5
145.4
151.8 139.0
136.6
133.5 139.6
92.4
91.9 92.9
148.2
145.1 151.2
Weight
* Difference from control significant (at 5% probability level). t Difference from control approaches significance (at 5% probability level). *** Difference from control significant (at 0.1% probability level).
Standard error of single pen means
Average
A+10% D.D.S.
Average
A+6% D.D.S.
Average
A+4% D.D.S.
Average
A+2% D.D.S.
Average
Equiv. to 10% D.D.S.
41.5 41.8
28.7*
70.0
41.2
A+Ash
Average
28.4 29.0
69.9 70.0
41.5 41.0
A B
13.8***
55.6
41.8
Average
A+Ash Equiv. to 3 % D.D.S.
39.4
79.2
39.8
12.5 15.0
38.0 40.7
78.0 80.4
40.0 39.7
Weight
Week 1
Weight increase
54.8 56.4
A B
A B
Initial weight
42.3 41.4
Control B (Low Manganese)
Average
Control A (High Manganese)
Brooder
5.9
119.6f
126.0 113.1
116.8
113.2 120.5
111.8
112.4 111.3
109.4
108.5 110.2
103.8
110.3 97.2
95.3f
92.0 98.6
50.6***
49.6 51.5
108.3
105.1 111.5
Weight increase
Week 2
0.061
1.386
1.334 1.438
1.400
1.407 1.392
1.415
1.442 1.388
1.456
1.459 1.453
1.550
1.514 1.586
1.564
1.639 1.490
2.440***
2.386 2.494
1.455
1.462 1.448
F.C.E.
1.727 1.622 1.674 1.615 1.571
373.2 369.7 371.4 380.8 371.9
416.8 412.0 414.4 422.7 413.1
1.460 1.440 1.450 1.460 1.395
227.3 229.4 228.4 235.9 228.8
270.9 271.7
10.1
238.2
14.9
391.Of
430.0 1.425 277.2
0.066
1.583 1.680 399.2 382.8 438.2 421.9 1.360 1.490 252.0 224.4
0.062
1.632
1.588 389.2 430.8 1.407 291.0 263.5
244.0t
1.596 1.579 387.0 391.4 428.3 433.2 1.431 1.383 237.6 250.4 278.9 292.2 285.6
1.593 376.4 417.9 1.428 232.4 273.9
277.8 270.0
271.3
1.704 352.8 394.4 1.533
1.716 1.693 364.3 341.2 405.8 383.0
1.826 321.7* 363.0
1.368 1.552 1.514
1.863 1.789 302.9 340.5
2.010*** 239.7*** 281.6 344.4 381.5
1.932 2.088
1.586 373.4 250.0 229.4
1.596 1.576
363.6 383.3
292.3 270.8
413.3
403.6 423.0
F.C.E.
1.265 1.470
1.933***
1.904 1.962
1.432
1.441 1.422
Weight
218.5
226.4 210.6
206.4f
200.7 212.2
132.9***
132.4 133.4
226.8
222.0 231.7
F.C.E.
Week 4 Weight increase
260.2
267.9 252.4
247.7
242.2 253.2
174.8
174.7 174.8
266.7
262.0 271.4
Weight
Weight increase
Week 3
TABLE 4.—Influence of D.D.S., D.D.S. ash and reduced manganese on growth, food conversion (Experiment 3). High manganese diets (Means of 25 chicks per sub-group, weight in grams F.C.E. g.food/g.gain)
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A B
A B
A + A s h e q u i v . to 3 % D.D.S.
A + A s h equiv. to 1 0 % D.D.S.
A B
A + 1 0 % D.D.S.
Control B (Low Manganese)
1.156 1.181
37.2
1.466 1.299 1.382
308.0 324.4 316.2
367.6 345.2 362.4
1.390 1.240 1.131
227.2
215.6 224.0
252.8 262.0
1.206 1.330 1.167
112.4 102.5 115.1
150.0 139.7 153.1
0.872 0.912 0.796
35.1 37.7
358.0 374.7
353.8 396.6 412.5
1.186 1.517 1.346
219.8
253.6 261.2
257.4 292.2 299.0
1.248 1.268 1.326
108.8 133.9 122.8
146.4 172.5 160.6
0.854 0.908 0.906
36.4»* 41.8 40.9
74.0
38.2
0.100
5.3
0.065
8.0
0.069
8.5
0.076
1.232* 362.4** 399.7 1.162** 267.7*** 305.0 1.158*
132.2**
169.4
0.568*
43.2***
80.6
37.3 1.3
1.220 1.243 405.6 393.8 1.090 1.234 277.0 258.4
1.374 351.8* 368.0 356.8
389.5
1.394 240.4* 314.6 295.4
278.1 1.179 1.138
129.8 134.5
167.4 171.5
0.460 0.675
41.8 44.7
1.178
116.2
153.8
0.774
37.4*
79.4 81.7
37.7
1.395 1.353
1.456
75.1
353.6 350.0
1.450 1.337 239.0 241.8 277.2 279.0
1.242 1.115
114.1 118.2
152.3 155.4
0.911 0.636
35.8 39.0
1.435 1.468
1.416
1.468 1.365
1.524
1.634 1.413
1.504
37.6 37.0
344.0
381.2 391.8 387.2
1.480 230.4 267.6
1.344
108.4
145.6
0.886
35.6
37.2
74.0 76.2
36.8 37.6
72.8
347.4 340.5
384.2 378.1 1.536 1.425
1.412 1.257
112.7 104.2
149.5 141.8
0.889 0.882
36.0 35.2
38.2 37.2
329.2
367.2 1.415 215.8 232.2 228.7
253.8 269.0 266.3
1.277
106.4
144.4
0.890
34.2
38.0
72.8 72.8
332.0 326.4
370.0 364.3 1.419 1.411
226.8 204.8
1.331 1.223
106.1 106.8
144.1 144.7
0.925 0.854
33.9 34.4
71.9 72.3
72.1
366.4**
404.6 1.432
257.4**
295.6 264.8 242.7
1.297
128.4**
166.6
0.907
41.4***
79.6
38.0 37.9
37.6
80.4 78.7
330.0
38.6 37.8
264.7
1.504 1.505
36.4"
1.486
72.3 75.7
37.2 38.0
324.4 335.7
74.0
361.4 373.7
1.393 1.386
37.5
323.1
360.2
1.312
222.9
112.8 112.1
149.8 150.1
0.875 0.868
263.8 265.6
1.181 1.230
109.9
147.0
0.810 226.8 227.6
1.439 1.533
F.C.E.
35.9 37.0
366.5 354.0
31.4
328.7 317.5
Weight
F.C.E. 1.339 1.285
223.7 222.1
72.9 75.0
260.0
Week 4 Weight increase
68.6
261.5 258.6
Weight
Week 3
Weight increase
37.0 38.0
1.168
F.C.E.
109.7 110.0
Weight 147.5 146.5
F.C.E. 0.780 0.841
29.3 33.5
67.1 70.0
Week 2
37.8 36.5
Weight
Week 1 Weight increase
Weight increase
Initial weights
* Difference from appropriate control significant (at 5% probability level). ** Difference from appropriate control significant (at 1% probability level). *** Difference from appropriate control significant (at 0.1 % probability level).
Standard error of single pen means
Average
B + 1 0 % D.D.S.
Average
A B
A B
B + A s h equiv. to 1 0 % D.D.S.
Average
A B
B + A s h equiv. to 3 % D.D.S.
Average
Average
A B
Average
Average
Average
A B
Control A (High Manganese)
Brooder
TABLE 5.—Influence of D.D.S. and D.D.S. ash on growth and food conversion using two different diets—high and low manganese
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528
W. P. JAFFE AND J. A. WAKELAM
sponse to the ash of D.D.S. seem well established. DISCUSSION
Our principal object in this work was to eliminate complicating effects due to the mineral content of the solubles so as to facilitate later fractionation work. To this extent we consider that the increased manganese level is justified. The true significance of these findings is more difficult to establish. Clearly the results of Experiments 3 and 4 are quite contradictory so far as the effect of manganese alone is concerned and we can offer no explanation for this. It seems unlikely in the extreme that the lower microbial loading of the gut, which we believe to be responsible for the poor response to D.D.S. in Experiment 3, should also have such a marked effect upon the requirement for manganese. If we assume that due to unknown causes the growth on diet 2 (low manganese control) is aberrant then we could see some measure of agreement with experiment 4—but we know of nothing to justify such an assumption. Turning to Experiment 4 we see that although the higher level of manganese does not itself influence growth in this experiment it appears to abolish any response due to the solubles ash. Analysis of the ash itself shows it to contain a fairly high level of manganese (138 p.p.m.) but apart from magnesium, calcium and potassium, no other mineral components seem to be present in appreciable amounts, bearing in mind the original level of minerals in the basal diet. It is possible that the relatively high level of calcium present in the soluble ash would disturb the calcium/phosphorus ratio and this might account for the depressions sometimes shown to the ash but this is difficult to reconcile with the fact that D.D.S. with the same level of calcium
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(8X2 Randomised Block) were: 1. Control A (Manganese content 185 mg./lb.) 2. Control A-fash equivalent to 3 % D.D.S. 3. Control A+ash equivalent to 10% D.D.S. 4. Control A + 1 0 % D.D.S. 5. Control B (Manganese content 54 mg./lb.) 6. Control B + ash equivalent to 3 % D.D.S. 7. Control B + ash equivalent to 10% D.D.S. 8. Control B + 10% D.D.S. Table 5 shows the detailed results. The following conclusions may be drawn: a. The comparison of the control diets is completely at variance with that in Experiment 3 since in this new experiment there is no difference between the growth rates on the two diets. The noticeably poorer growth during week 1 on diet 1 in brooder A makes this figure appear discrepant and it is probably best to ignore comparisons at this week. b. The lower level of ash produces small responses which nowhere approach significance (ignoring week 1) on the high manganese diet. Although the responses on the low manganese control diet are considerably higher they still fail to reach significance. c. The higher level of ash depresses growth slightly on the high manganese diets (again ignoring week 1) but gives a good response on the low manganese diet, reaching statistical significance at weeks 3 and 4. d. The 10% D.D.S. response is highly significant ( 1 % level or better) at all weeks and on both levels of manganese. Whilst it is difficult to explain why there is no difference between the two control diets in this experiment the effect of added manganese in abolishing any re-
GROWTH FACTORS IN MALT DISTILLERS DRIED SOLUBLES
SUMMARY
1. Using a semi-synthetic diet responses in terms of growth and food conversion efficiency have been obtained to malt distillers dried solubles. 2. The ash of these solubles also gave responses in terms of growth under certain circumstances but this response was abolished by raising the manganese content of the basal diet from 54 mg./lb. to 185 mg./lb. 3. Although this indicated manganese as a growth factor in solubles ash, and analysis revealed relatively high levels of manganese in the ash, certain features of the results cast doubt on this simple explanation. ACKNOWLED GEMENTS
The assistance of The Agricultural Research Council and The Distillers Company (Biochemicals) Limited in bearing part of the cost of this investigation and of The Boots Pure Drug Company Limited, in providing the mineral mixtures used, is
gratefully acknowledged. Thanks are due to Mr. J. Ince of The Distillers Company Limited for carrying out the statistical computations. REFERENCES Bird, H. R., H. J. Almquist, W. W. Cravens, F. W. Hill and J. McGinnis, 1954, Nutrient requirements for poultry: National Research Council Publication 301. Briggs, G. M., 1956. Inadequacy of certain salt mixtures used in studies of unidentified growth factors for chicks. Poultry Sci: 35: 740-742. Coates, M. E., C. D. Dickinson, C. F. Harrison, S. K. Kon, J. W. G. Porter, S. H. Cummings and W. F. J. Cuthbertson, 1952. Antibiotics in chick nutrition. J. Sci. Food Agr. 3: 43-48. Combs, C. F., C. B. Sweet, H. J. Jones, C. L. Romoser and R. W. Bishop, 1954. Multiplicity of unidentified growth factors required by chicks and poults. Poultry Sci.: 33: 1050. Couch, J. R., B. L. Reid, A. A. Camp, W. N. Dannenburg and E. E. Rozacky, 1955. Distillers dried solubles in the diet of chicks. Proc. 10th Distillers Feed Conf. (Cincinnati): 47-57. Jaffe, W. P., and J. A. Wakelam, 1956. Unidentified chick growth factors in fermentation byproducts. Nature (London), 178: 414. Morrison, A. B., M. L. Scott and L. C. Norris, 1955. Evidence for an unidentified mineral required by the chick. Poultry Sci.: 34: 738-740. Norris, L. C. 1955. Recent progress in unidentified growth factors in distillers dried solubles and other feed supplements. Proc. 10th Distillers Feed Conf. (Cincinnati): 39-42. Rasmussen, R. A., P. W. Luthy, J. M. Van Lanen and C. S. Borult, 1957. Measurement and differentiation of unidentified chick growth factors using a new semi-purified ration. Poultry Sci. 36:46-54. Scott, M. L., 1957. Corn distillers dried solubles as a source of unidentified nutrients required by chickens and turkeys. Proc. 12th Distillers Feed Conf. (Cincinnati): 61-70. Wakelam, J. A., and W. P. Jaffe, 1958. Growth promoting factors in malt distillers dried solubles: Field trials and preliminary findings. Brit. J. Nutr. 12: 147.
JULY 22-24. AMERICAN POULTRY HATCHERY FEDERATION CONVENTION, AUDITORIUM, CLEVELAND, OHIO
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does not do so or that this effect is more marked in the presence of additional manganese. Although there are obvious discrepancies, and the amount of manganese which the solubles ash provides in the final ration is relatively small it is difficult to find any explanations for the facts which we report here other than that manganese is acting as a growth promoting factor in the ash of malt distillers solubles. So far as we are concerned, our main object has been to eliminate responses to the mineral portion of the solubles from the present, and this has been acheived by increasing the level of manganese in the basal diet.
529
HE value of corn distillers solubles in the diets of young stock is well established and they are standard ingredients in many American feeds. In addition to the protein, energy and vitamins which they provide, there is strong evidence that they contain also an unidentified growth factor for chicks and poults. This evidence has been the subject of several adequate reviews (Combs et al., 1954; Scott, 1957; Rasmussen et al. 1957; and others) and it will not, therefore, be further reviewed here. In essence, however, the present position seems to be somewhat confused as to the identity or otherwise of the factor(s) provided by fish meals and by-products, milk products, yeast, corn distillers solubles and other fermentation solubles. Several workers have suggested that part, at least, of the growth promoting properties of these various products derives from their mineral components but here again the situation is somewhat obscure. Morrison et al. (1955) and Norris (1955) observed growth responses approximately half as great as those to the intact products when feeding the ash of five supposed growth factor sources. Dannenburg et al. (1955) and Couch (1955) obtained a similar increase when feeding the ash equivalent to 3 percent corn distillers solubles. Briggs (1956)
has criticized the basal diets used in some of these experiments for falling short of the minimum levels laid down by the National Research Council (Bird et al., 1954), and states that he has failed to establish any growth response to the ash of a number of feedstuffs when added to a purified diet. Scott et al. (1955) also failed to secure responses to the ash of corn distillers solubles. Thus it seems that the experiences of different workers with the mineral components of the various supposed growth factor sources has been as confusing as with the intact products themselves. We have discussed this general point in relation to our experience with malt distillers solubles elsewhere (Wakelam and Jaffe, 1958) and are here concerned principally with our work on malt distillers solubles ash. The recovery of malt distillers solubles from the Scottish distilleries is still at an early stage in contrast to the corn distillers solubles which have been recovered ever since the repeal of Prohibition. Accordingly there is little evidence concerning their properties. We have presented certain preliminary findings (Wakelam and Jaffe, 1958; Jaffe and Wakelam, 1956) and in the former have explained the essential differences between the malt and corn distillers' prod-
520
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T
GROWTH FACTORS IN MALT DISTILLERS DRIED SOLUBLES
these several reasons we do not suggest that our results are necessarily relevant to those obtained with corn distillers solubles mentioned above. EXPERIMENTAL METHODS
The basal diet which has been used in this work is shown in Table 1. This diet was designed to support a high level of growth whilst yet being capable of improvement from the addition of distillers solubles—which term, throughout this paper, refers to the recovered solubles from the "pot-stills" of a malt distillery in the Scottish Highlands. The casein and gelatin are standard edible grade material as also is the maize and starch and all
TABLE 1.—Standard diet for growth-factor work on chicks Basal diet Casein Gelatin Maize starch Sucrose Lard Distillers dried grains Mineral mix
Additions 25% 15% 29% 10% 10% 5% 6%
Any additions were made at the expense of maize starch
Mineral CaC0 3 (%) Steamed bone flour (22% Ca 17% P) (%) CaH4-(P04)2H20(%) KC1 (%) NaCl(%) MgS0 4 (%) MnSCv4H20(%) ZnCl2 (g./100-lb.) K I (g./100-lb.) H3BO3 (g./100-lb.) CoS0 4 -7H 2 0 (g./100-lb.) CuSO 4 -5H 2 O(g./100-lb.) FeSO 4 -7H 2 O(g./100-lb.)
Cystine (%) Vitamin A (I.U./lb.) Cholecalciferol (I.U./lb.) a-tocopheryl acetate (mg./lb.) Menaphthone (mg./lb.) Cyanocobalamin (mg./lb.) Thiamine hydrochloride (mg./lb.) Riboflavin (mg./lb.) Calcium pantothenate (mg./lb.) Nicotinic acid (mg./lb.) Choline chloride (mg./lb.) Pyridoxine hydrochloride (mg./lb.) Biotin (mg./lb.) ^>-aminobenzoic acid (mg./lb.) Inositol (mg./lb.) Folic acid (mg./lb.) Procaine benzyl penicillin (mg./lb.) »••• i. • . Mineral mix amount 9.55) 50.00J 6.60\ / 10.42 12.22 6.38 2.78 12 25 6 6 67 880
0.25 4,000 1,000 4.5 2.0 0.008 1.6 2.6 8.4 24.0 1,200.0 2.6 0.08 2.6 450.0 0.5 4.0
Contribution to basal diet when a(Wed at
Ca (%) P(%) Ratio Ca:P K(%) NaCl(%) Mg (mg./lb.) Mn (mg./lb.) Zn (mg./lb.) I (mg./lb.) B (mg./lb.) Co (mg./lb.) Cu (mg./lb.) Fe (mg./lb.)
6 %
1.005 1.005 1.67 0.33 0.73 225 185 3.45 11 0.8 0.8 12 120
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ucts. It should be made clear at this stage that in our studies of the properties of the ashed material we were concerned primarily to exclude the effect of the ash from our work. This, of course, is in contrast to the approach of the workers already cited who were interested in the ash per se. It is our intention to return to the study of the ash at a later stage but at present we use in our basal diet semi-purified ingredients and the birds receive ordinary tap water—no attempt is made to exclude unknown minerals, rather have we sought in formulating our basal diet to include all possible nutrients so long as the addition of malt distillers solubles will still elicit a growth response. For
521
522
W. P. JAFFE AND J. A. WAKELAM
The diets were made up at frequent intervals (usually about one week according to rate of consumption) and were kept in a cold store. These precautions were considered advisable because of the high level of fat included in the basal diet. RESULTS
Experiment 1 was a 4X4 factorial experiment with the following treatments: 1. Control as in Table 1 2. Control+3% spray-dried fish solubles
3. Control+6% malt distillers dried solubles (hereafter referred to as D.D.S.) 4. Control+ash equivalent to 6% D.D.S: Table 2 shows the results in terms of both growth and food conversion efficiency (F.C.E.). The following conclusions may be drawn: a. A good growth response to D.D.S. which is statistically significant (5% level) at 2, 3, 4 weeks. A marked F.C.E. response to D.D.S. which, however, does not reach a significance. b. D.D.S. ash produces a small growth response which at no stage approaches significance—it is at all stages well under half that to D.D.S. The F.C.E. response is even smaller and is in fact negative in the first week. c. The growth response to fish solubles is very small—in most weeks less than to D.D.S. ash. The F.C.E. response is more marked especially at week 4 where it reaches significance. Experiment Z was an 8X2 Randomised Block arrangement with the following treatments: 1. Control as Table 1 but manganese level reduced to 54 mg./lb. 2. Control+ash equivalent to 3 % D.D.S. 3. Control+ash equivalent to 10% D.D.S. 4. Control+1% D.D.S. 5. Control+2% D.D.S. 6. Control+4% D.D.S. 7. Control+6% D.D.S. 8. Control+10% D.D.S. Table 3 shows the results in detail. The mean weights for treatment 2 in brooder B are unusually low and as this was believed to be due to a faulty heater they have been replaced in the analysis by values estimated by minimising the sums of squares for TreatmentsX Brooders. These estimates are shown in brackets in
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these and the dried grains, added to provide roughage, will naturally provide some minerals. It was hoped that by the high level of protein used we would avoid any effect due to the protein content of the solubles. The manganese level shown in Table 1 is that now included as standard—a lower level was used in some of the experiments here reported. The experimental unit comprises two four-tier electrically heated brooders with thermostatic control. Each wire-floored tier is split longitudinally providing sixteen separate compartments. The chicks used were ordinary commercial hatchery stock (Rhode IslandXLight Sussex) which were allocated at random to the sixteen compartments on arrival as day olds. In Experiment 1 of this paper pullets were used, in all other experiments cockerels. All birds were weighed individually on arrival and at weekly intervals for four weeks and both feed and water were available ab libitum. Food consumption was measured on a group basis. No allowance was made for wastage as it had been found that by reducing the amount of food available this could be virtually eliminated. After a few experiments with 4X4 Latin Square layouts it was established that there was no significant effect due to position and subsequent experiments have been of the 8X2 Randomised Block design.
523
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504
F.C.E.
G R O W T H FACTORS IN M A L T D I S T I L L E R S D R I E D SOLUBLES
t * ** ***
29.6 (33.2)t 40.9 40.0 40.4** 32.8 31.8 32.3 34.3 30.9
68.8
79.5 78.9
79.2
70.8 69.9
70.4
72.8 68.8
39.1
38.6 38.9
38.8
38.0 38.1
38.0
38.5 37.9
A B
A B
A B
A B
A B
A B
36.9 38.3 37.6* 46.4 37.5 42.0*** 44.1 46.9
75.2
84.6 76.1
80.4
82.5 84.5
83.5
37.6
38.2 38.6
38.4
38.4 37.6
38.0 2.7
45.5***
32.6
70.8
73.9 76.4
38.2
37.0 38.1
27.0
151.8 148.6
1.049 1.103
Week 2
1.515** 1.496 1.571
357.5** 371.0 342.7
395.6 409.5 380.6
1.466 1.450 1.504
206.3*
1.534** 1.515 1.560 1.538** 1.531 1.533
356.8** 378.9 387.7 383.3*** 387.2 400.8
395.0
420.8 425.4 439.4
1.430' 1.489 1.421
226.1** 221.4 239.5
263.6 259.6 278.1
0.073
2.5
0.050
8.8
0.059
9.9
406.4***
444.4 1.437* 246.0*** 284.0 1.362* 126.1***
164.1
0.026
1.542**
1.534 1.551 409.6 4C3.2 448.0 440.8 1.402 1.472 255.6 236.5
294.0 274.1 1.371 1.354 127.2 125.0
165.6 162.6
0.987*
1.532** 394.0*** 432.4 1.455* 230.5**
268.8 1.376*
118.0***
156.4
1.477
415.9 425.8
209.0*
1.404 1.456
247.2
222.8 229.3
215.1 202.9
1.532 1.498 362.1 352.9 400.1 391.0
390.2
1.630*
1.629 1.631 348.5 354.4 387.1 393.3
351.4**
2.158** 244.4 (277.0)t*
317.4 249.7 283.6
1.690
1.663 1.716
2.146 (2.169)
312.2
313.4 311.0
278.3 210.6 (275.7)t
353.0
354.3 351.8
F.C.E.
1.522 1.411
1.428*
1.389 1.468
1.895*
1.896 (1.894)
1.634
1.680 1.589
Weight
206.5 206.1
233.0***
234.3 231.8
148.9 (167.l)f
167.4 130.4 (166.8)t
176.7
173.9 179.5
F.C.E.
Week 4 Weight increase
259.8 267.4
253.6 240.8
244.4
244.5 244.2
271.8
272.9 270.7
188.0
206.5 169.5
217.6
214.8 220.3
Weight
Week 3
Weight increase
1.026 0.948
1.355 1.398
1.435*
106.2*** 120.4 115.6
1.400 1.469
1.530
1.483 1.576
1.489
1.528 1.450
1.407*
1.338 1.476
1.726*
1.707 (1.746)
1.566
1.552 1.581
F.C.E.
104.3 108.1
97.1*
100.4 93.8
94.2
94.3 94.0
111.4***
113.2 109.7
75.2 (81.8)t
82.8 67.6 (80.9)t
84.9
84.8 85.0
Weight increase
diet
1.070
158.6 154.2
143.8
1.127 1.000 1.141
141.3 146.2
135.3
138.9 131.7
132.2
132.3 132.1
1.107 1.147
1.312
1.212 1.413
1.246
1.285 1.206
150.2
114.3
1.264
1.076
121.9 106.7
1.241 1.288
125.8
73.1 64.4
39.1 39.1
1.264
34.0 25.3 (32.5)t
67.8
40.9
A B
125.7 125.8
1.238 1.290
27.2 26.7
68.1 67.5
40.9 40.8
Weight
F.C.E.
A B
Weight
Week 1 Weight increase
Initial weight
Means of brooder
Estimated values—see text, Difference from control significant a t 5% probability level. Difference from control significant a t 1 % probability level. Difference from control significant at 0 . 1 % probability level.
Standard error of single pen means
Average
10% D.D.S.
Average
6 % D.D.S.
Average
4 % D.D.S.
Average
2 % D.D.S.
Average
1% D.D.S.
Average
Ash equiv. to 1 0 % D.D.S.
Average
Ash Equiv. to 3 % D.D.S.
Average
Control
Diet
TABLE 3.—Influence of different levels of D.D.S. and D.D.S. ash on growth and food conversion {Experiment 2). Low (Means of 25 chicks per sub-group, weights in grams, food conversion in g.food/g.weight increase)
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GROWTH FACTORS IN MALT DISTILLERS DRIED SOLUBLES
7. Control A + 6 % D.D.S. 8. Control A + 1 0 % D.D.S. Table 4 shows the detailed results from which the following conclusions may be drawn: a. Control A (high manganese) gave considerably greater growth rates and superior F.C.E. than Control B (low manganese). The difference for both growth and conversion were highly significant (0.1% level of probability) at all weeks. b. Both levels of ash appeared to depress growth in this experiment. In the case of the lower level (3% ash equivalent) this depression was statistically significant at 1 and 4 weeks. c. The response to D.D.S. although reasonably consistent approached statistical significance only for the higher levels (6% and 10%) and in the later weeks. At no stage was the response significant at the 5% level of probability. These results were somewhat surprising. Between Experiments 2 and 3 of this paper the brooders had been moved to a new building and this poor response to D.D.S. led us to consider whether the solubles were acting through the gut flora and we were experiencing a "clean/dirty" house effect—comparable to that reported by Coates et al. (1952). This point has been investigated and is reported elsewhere (Wakelam and Taffe, 1958) where we have discussed the matter in some detail and suggested that it may indeed be the case. Since this suggests that Experiment 3 may represent an atypical state of affairs, we deferred confirmation until we felt reasonably certain that the new house had reached a steady state, and then carried out Experiment 4. In Experiment 4 we examined the effect of two levels of D.D.S. ash and a high level of D.D.S. on the two control diets used in Experiment 3. The treatments
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Table 3. The following conclusions may be drawn. a. 10% ash equivalent produces growth responses which are statistically significant ( 1 % level or better) at all weeks. The F.C.E. response is also significant (5% level or better) at all weeks. b. The response to 10% D.D.S. is always considerably greater than that to the equivalent amount of ash and the difference between ash and solubles response is significant (5% level or better) for both growth and conversion at 2 and 4 weeks. c. 3 % ash equivalent shows growth depression and an adverse effect on conversion at all weeks. The effect is significant (5% level) at 4 weeks. d. It is estimated, from the regression equation, that the growth response to 3 % D.D.S. would be considerably greater than that to 3 % ash equivalent at all weeks and that it would be statistically significant (5% level). These results seem to indicate a marked response to a high level of D.D.S. ash and are in many respects more similar to those reported by the various American workers quoted above than to the results of Experiment 1. Experiment 3 was essentially a repeat of Experiment 2 with the manganese level restored to the high value of Table 1 and an additional group (treatment 2) receiving the same diet as the control in Experiment 2. The treatments (8X2 Randomised Block) were: 1. Control A (Manganese content 185 mg./lb.) 2. Control B (Manganese content 54 mg./lb.) 3. Control A+ash equivalent to 3 % D.D.S. 4. Control A+ash equivalent to 10% D.D.S. 5. Control A + 2 % D.D.S. 6. Control A + 4 % D.D.S.
525
A B
A B
A B
A B
A B
40.4
79.4
39.0
3.1
37.8
41.6
43.7 37.1
34.7 41.0
76.0 82.8
41.3 41.8
79.4
36.6
78.2
41.6
82.7 76.2
36.1 37.1
39.0 39.1
35.4
43.0
78.0 78.3
34.9 35.8
78.5 78.1
43.6 42.3
78.3
34.2
75.8
41.6
41.9 41.2
37.2 31.2
78.7 73.0
0.113
1.093
0.992 1.193
1.096
1.108 1.083
1.098
1.121 1.076
1.163
1.174 1.152
1.233
1.187 1.279
1.434
1.530 1.337
2.286***
2.466 2.107
1.080
1.121 1.040
F.C.E.
158.6
165.0 152.2
158.4
154.5 162.3
153.4
154.3 152.5
152.3
152.1 152.5
145.4
151.8 139.0
136.6
133.5 139.6
92.4
91.9 92.9
148.2
145.1 151.2
Weight
* Difference from control significant (at 5% probability level). t Difference from control approaches significance (at 5% probability level). *** Difference from control significant (at 0.1% probability level).
Standard error of single pen means
Average
A+10% D.D.S.
Average
A+6% D.D.S.
Average
A+4% D.D.S.
Average
A+2% D.D.S.
Average
Equiv. to 10% D.D.S.
41.5 41.8
28.7*
70.0
41.2
A+Ash
Average
28.4 29.0
69.9 70.0
41.5 41.0
A B
13.8***
55.6
41.8
Average
A+Ash Equiv. to 3 % D.D.S.
39.4
79.2
39.8
12.5 15.0
38.0 40.7
78.0 80.4
40.0 39.7
Weight
Week 1
Weight increase
54.8 56.4
A B
A B
Initial weight
42.3 41.4
Control B (Low Manganese)
Average
Control A (High Manganese)
Brooder
5.9
119.6f
126.0 113.1
116.8
113.2 120.5
111.8
112.4 111.3
109.4
108.5 110.2
103.8
110.3 97.2
95.3f
92.0 98.6
50.6***
49.6 51.5
108.3
105.1 111.5
Weight increase
Week 2
0.061
1.386
1.334 1.438
1.400
1.407 1.392
1.415
1.442 1.388
1.456
1.459 1.453
1.550
1.514 1.586
1.564
1.639 1.490
2.440***
2.386 2.494
1.455
1.462 1.448
F.C.E.
1.727 1.622 1.674 1.615 1.571
373.2 369.7 371.4 380.8 371.9
416.8 412.0 414.4 422.7 413.1
1.460 1.440 1.450 1.460 1.395
227.3 229.4 228.4 235.9 228.8
270.9 271.7
10.1
238.2
14.9
391.Of
430.0 1.425 277.2
0.066
1.583 1.680 399.2 382.8 438.2 421.9 1.360 1.490 252.0 224.4
0.062
1.632
1.588 389.2 430.8 1.407 291.0 263.5
244.0t
1.596 1.579 387.0 391.4 428.3 433.2 1.431 1.383 237.6 250.4 278.9 292.2 285.6
1.593 376.4 417.9 1.428 232.4 273.9
277.8 270.0
271.3
1.704 352.8 394.4 1.533
1.716 1.693 364.3 341.2 405.8 383.0
1.826 321.7* 363.0
1.368 1.552 1.514
1.863 1.789 302.9 340.5
2.010*** 239.7*** 281.6 344.4 381.5
1.932 2.088
1.586 373.4 250.0 229.4
1.596 1.576
363.6 383.3
292.3 270.8
413.3
403.6 423.0
F.C.E.
1.265 1.470
1.933***
1.904 1.962
1.432
1.441 1.422
Weight
218.5
226.4 210.6
206.4f
200.7 212.2
132.9***
132.4 133.4
226.8
222.0 231.7
F.C.E.
Week 4 Weight increase
260.2
267.9 252.4
247.7
242.2 253.2
174.8
174.7 174.8
266.7
262.0 271.4
Weight
Weight increase
Week 3
TABLE 4.—Influence of D.D.S., D.D.S. ash and reduced manganese on growth, food conversion (Experiment 3). High manganese diets (Means of 25 chicks per sub-group, weight in grams F.C.E. g.food/g.gain)
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A B
A B
A + A s h e q u i v . to 3 % D.D.S.
A + A s h equiv. to 1 0 % D.D.S.
A B
A + 1 0 % D.D.S.
Control B (Low Manganese)
1.156 1.181
37.2
1.466 1.299 1.382
308.0 324.4 316.2
367.6 345.2 362.4
1.390 1.240 1.131
227.2
215.6 224.0
252.8 262.0
1.206 1.330 1.167
112.4 102.5 115.1
150.0 139.7 153.1
0.872 0.912 0.796
35.1 37.7
358.0 374.7
353.8 396.6 412.5
1.186 1.517 1.346
219.8
253.6 261.2
257.4 292.2 299.0
1.248 1.268 1.326
108.8 133.9 122.8
146.4 172.5 160.6
0.854 0.908 0.906
36.4»* 41.8 40.9
74.0
38.2
0.100
5.3
0.065
8.0
0.069
8.5
0.076
1.232* 362.4** 399.7 1.162** 267.7*** 305.0 1.158*
132.2**
169.4
0.568*
43.2***
80.6
37.3 1.3
1.220 1.243 405.6 393.8 1.090 1.234 277.0 258.4
1.374 351.8* 368.0 356.8
389.5
1.394 240.4* 314.6 295.4
278.1 1.179 1.138
129.8 134.5
167.4 171.5
0.460 0.675
41.8 44.7
1.178
116.2
153.8
0.774
37.4*
79.4 81.7
37.7
1.395 1.353
1.456
75.1
353.6 350.0
1.450 1.337 239.0 241.8 277.2 279.0
1.242 1.115
114.1 118.2
152.3 155.4
0.911 0.636
35.8 39.0
1.435 1.468
1.416
1.468 1.365
1.524
1.634 1.413
1.504
37.6 37.0
344.0
381.2 391.8 387.2
1.480 230.4 267.6
1.344
108.4
145.6
0.886
35.6
37.2
74.0 76.2
36.8 37.6
72.8
347.4 340.5
384.2 378.1 1.536 1.425
1.412 1.257
112.7 104.2
149.5 141.8
0.889 0.882
36.0 35.2
38.2 37.2
329.2
367.2 1.415 215.8 232.2 228.7
253.8 269.0 266.3
1.277
106.4
144.4
0.890
34.2
38.0
72.8 72.8
332.0 326.4
370.0 364.3 1.419 1.411
226.8 204.8
1.331 1.223
106.1 106.8
144.1 144.7
0.925 0.854
33.9 34.4
71.9 72.3
72.1
366.4**
404.6 1.432
257.4**
295.6 264.8 242.7
1.297
128.4**
166.6
0.907
41.4***
79.6
38.0 37.9
37.6
80.4 78.7
330.0
38.6 37.8
264.7
1.504 1.505
36.4"
1.486
72.3 75.7
37.2 38.0
324.4 335.7
74.0
361.4 373.7
1.393 1.386
37.5
323.1
360.2
1.312
222.9
112.8 112.1
149.8 150.1
0.875 0.868
263.8 265.6
1.181 1.230
109.9
147.0
0.810 226.8 227.6
1.439 1.533
F.C.E.
35.9 37.0
366.5 354.0
31.4
328.7 317.5
Weight
F.C.E. 1.339 1.285
223.7 222.1
72.9 75.0
260.0
Week 4 Weight increase
68.6
261.5 258.6
Weight
Week 3
Weight increase
37.0 38.0
1.168
F.C.E.
109.7 110.0
Weight 147.5 146.5
F.C.E. 0.780 0.841
29.3 33.5
67.1 70.0
Week 2
37.8 36.5
Weight
Week 1 Weight increase
Weight increase
Initial weights
* Difference from appropriate control significant (at 5% probability level). ** Difference from appropriate control significant (at 1% probability level). *** Difference from appropriate control significant (at 0.1 % probability level).
Standard error of single pen means
Average
B + 1 0 % D.D.S.
Average
A B
A B
B + A s h equiv. to 1 0 % D.D.S.
Average
A B
B + A s h equiv. to 3 % D.D.S.
Average
Average
A B
Average
Average
Average
A B
Control A (High Manganese)
Brooder
TABLE 5.—Influence of D.D.S. and D.D.S. ash on growth and food conversion using two different diets—high and low manganese
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528
W. P. JAFFE AND J. A. WAKELAM
sponse to the ash of D.D.S. seem well established. DISCUSSION
Our principal object in this work was to eliminate complicating effects due to the mineral content of the solubles so as to facilitate later fractionation work. To this extent we consider that the increased manganese level is justified. The true significance of these findings is more difficult to establish. Clearly the results of Experiments 3 and 4 are quite contradictory so far as the effect of manganese alone is concerned and we can offer no explanation for this. It seems unlikely in the extreme that the lower microbial loading of the gut, which we believe to be responsible for the poor response to D.D.S. in Experiment 3, should also have such a marked effect upon the requirement for manganese. If we assume that due to unknown causes the growth on diet 2 (low manganese control) is aberrant then we could see some measure of agreement with experiment 4—but we know of nothing to justify such an assumption. Turning to Experiment 4 we see that although the higher level of manganese does not itself influence growth in this experiment it appears to abolish any response due to the solubles ash. Analysis of the ash itself shows it to contain a fairly high level of manganese (138 p.p.m.) but apart from magnesium, calcium and potassium, no other mineral components seem to be present in appreciable amounts, bearing in mind the original level of minerals in the basal diet. It is possible that the relatively high level of calcium present in the soluble ash would disturb the calcium/phosphorus ratio and this might account for the depressions sometimes shown to the ash but this is difficult to reconcile with the fact that D.D.S. with the same level of calcium
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(8X2 Randomised Block) were: 1. Control A (Manganese content 185 mg./lb.) 2. Control A-fash equivalent to 3 % D.D.S. 3. Control A+ash equivalent to 10% D.D.S. 4. Control A + 1 0 % D.D.S. 5. Control B (Manganese content 54 mg./lb.) 6. Control B + ash equivalent to 3 % D.D.S. 7. Control B + ash equivalent to 10% D.D.S. 8. Control B + 10% D.D.S. Table 5 shows the detailed results. The following conclusions may be drawn: a. The comparison of the control diets is completely at variance with that in Experiment 3 since in this new experiment there is no difference between the growth rates on the two diets. The noticeably poorer growth during week 1 on diet 1 in brooder A makes this figure appear discrepant and it is probably best to ignore comparisons at this week. b. The lower level of ash produces small responses which nowhere approach significance (ignoring week 1) on the high manganese diet. Although the responses on the low manganese control diet are considerably higher they still fail to reach significance. c. The higher level of ash depresses growth slightly on the high manganese diets (again ignoring week 1) but gives a good response on the low manganese diet, reaching statistical significance at weeks 3 and 4. d. The 10% D.D.S. response is highly significant ( 1 % level or better) at all weeks and on both levels of manganese. Whilst it is difficult to explain why there is no difference between the two control diets in this experiment the effect of added manganese in abolishing any re-
GROWTH FACTORS IN MALT DISTILLERS DRIED SOLUBLES
SUMMARY
1. Using a semi-synthetic diet responses in terms of growth and food conversion efficiency have been obtained to malt distillers dried solubles. 2. The ash of these solubles also gave responses in terms of growth under certain circumstances but this response was abolished by raising the manganese content of the basal diet from 54 mg./lb. to 185 mg./lb. 3. Although this indicated manganese as a growth factor in solubles ash, and analysis revealed relatively high levels of manganese in the ash, certain features of the results cast doubt on this simple explanation. ACKNOWLED GEMENTS
The assistance of The Agricultural Research Council and The Distillers Company (Biochemicals) Limited in bearing part of the cost of this investigation and of The Boots Pure Drug Company Limited, in providing the mineral mixtures used, is
gratefully acknowledged. Thanks are due to Mr. J. Ince of The Distillers Company Limited for carrying out the statistical computations. REFERENCES Bird, H. R., H. J. Almquist, W. W. Cravens, F. W. Hill and J. McGinnis, 1954, Nutrient requirements for poultry: National Research Council Publication 301. Briggs, G. M., 1956. Inadequacy of certain salt mixtures used in studies of unidentified growth factors for chicks. Poultry Sci: 35: 740-742. Coates, M. E., C. D. Dickinson, C. F. Harrison, S. K. Kon, J. W. G. Porter, S. H. Cummings and W. F. J. Cuthbertson, 1952. Antibiotics in chick nutrition. J. Sci. Food Agr. 3: 43-48. Combs, C. F., C. B. Sweet, H. J. Jones, C. L. Romoser and R. W. Bishop, 1954. Multiplicity of unidentified growth factors required by chicks and poults. Poultry Sci.: 33: 1050. Couch, J. R., B. L. Reid, A. A. Camp, W. N. Dannenburg and E. E. Rozacky, 1955. Distillers dried solubles in the diet of chicks. Proc. 10th Distillers Feed Conf. (Cincinnati): 47-57. Jaffe, W. P., and J. A. Wakelam, 1956. Unidentified chick growth factors in fermentation byproducts. Nature (London), 178: 414. Morrison, A. B., M. L. Scott and L. C. Norris, 1955. Evidence for an unidentified mineral required by the chick. Poultry Sci.: 34: 738-740. Norris, L. C. 1955. Recent progress in unidentified growth factors in distillers dried solubles and other feed supplements. Proc. 10th Distillers Feed Conf. (Cincinnati): 39-42. Rasmussen, R. A., P. W. Luthy, J. M. Van Lanen and C. S. Borult, 1957. Measurement and differentiation of unidentified chick growth factors using a new semi-purified ration. Poultry Sci. 36:46-54. Scott, M. L., 1957. Corn distillers dried solubles as a source of unidentified nutrients required by chickens and turkeys. Proc. 12th Distillers Feed Conf. (Cincinnati): 61-70. Wakelam, J. A., and W. P. Jaffe, 1958. Growth promoting factors in malt distillers dried solubles: Field trials and preliminary findings. Brit. J. Nutr. 12: 147.
JULY 22-24. AMERICAN POULTRY HATCHERY FEDERATION CONVENTION, AUDITORIUM, CLEVELAND, OHIO
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does not do so or that this effect is more marked in the presence of additional manganese. Although there are obvious discrepancies, and the amount of manganese which the solubles ash provides in the final ration is relatively small it is difficult to find any explanations for the facts which we report here other than that manganese is acting as a growth promoting factor in the ash of malt distillers solubles. So far as we are concerned, our main object has been to eliminate responses to the mineral portion of the solubles from the present, and this has been acheived by increasing the level of manganese in the basal diet.
529