Growth-Promoting Effects of Fermented Soybeans for Broilers1

Growth-Promoting Effects of Fermented Soybeans for Broilersl C . C. C H A H , 2 C. W . CARLSON, 2 G. SEMENIUK, 3 I. S. PALMER 4 AND C. W . HESSELTINE 5

Departments of Animal Science,2 Plant Science,3 and Station Biochemistry,4 South Dakota State University, Brookings, South Dakota 57006 and Fermentation Laboratory,5 Northern Regional Research Laboratory, U.S.D.A., Peoria, Illinois 61604 (Received for publication July 30, 1974)

POULTRY SCIENCE 54: 600-609, 1975

INTRODUCTION

T

HE effects of fungi are both beneficial and detrimental. Fungi of some kind participate in many biological and chemical reactions which facilitate the production of food products that are consumed by humans. Many beverages, cheeses, antibiotics and certain kinds of candies are produced or altered by the help of fungi. On the other hand, there are fungi which have detrimental effects. After the report of heavy mortality among turkey poults from mycotoxins in England during the sixties (Blount, 1961), there have been extensive investigations on the detrimental effects of feeding molded feedstuffs to poultry and other farm animals. Various investigators have studied the effects of afla-

1. Approved for publication by the Director, Agricultural Experiment Station, South Dakota State University, Brookings, as Journal series No. 1283. Reference to a company or product name does not imply approval or recommendation of the product by the U.S.D.A. to the exclusion of others that may be suitable.

toxins in depressing chick growth and feed conversion (Richardson et al, 1962; Chute et al., 1965; Smith and Hamilton, 1970; Semeniuk et al, 1970; Sharby et al, 1973) and in reducing egg production and egg size (Sims et al, 1970; Hamilton and Garlich, 1971; Garlich et al., 1972). Aflatoxins have also been cited as the causative agents for the hemorrhagic syndrome in poultry (Forgacs, 1966; Allcroft, 1969). Several other mycotoxins have been characterized and their toxicological effects have also been evaluated in turkey poults and in chickens (Scott and Somers, 1969; Meronuck et al., 1970; Speers et al., 1971). Ochratoxin produced by Aspergillus ochraceus has been shown to be a potent mycotoxin for broiler chickens (Doupnik and Peckham, 1970; Tucker and Hamilton, 1971) and for laying hens (Choudhury et al., 1971). Wyatt and Hamilton (1972) presented data indicating that rubratoxin (from Penicillum rubrum) may be poisonous to poultry if fed at excessive levels. Surprisingly, very few data have been published on the beneficial role of fungi in poultry. Recent investigations in our labora-

600

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ABSTRACT Four factorial experiments are here reported on the use of control and fermented soybeans in glucose-soybean diets for broiler chicks with varying protein levels (15, 17 and 19% and 13, 16 and 19%). Eleven strains of Aspergilli were used as cultures in these experiments, selected on the basis of a prior history of beneficial effects. Data obtained from 4-week growth studies revealed that feeding soybeans fermented with 10 of the 11 species gave significant (P < 0.05) improvements in weight gain and feed efficiency. The responses were more pronounced with the low dietary levels of protein. No detrimental effect was shown from any of the cultures tested. Chemical analyses indicate that chicks fed the fermented soybean diets made better use of dietary nitrogen and dry matter. Carcass composition data show that the diets made with fermented soybeans produced chicks that were significantly (P < 0.05) higher in protein and ash and lower in total lipids. Amino acid analyses suggest that the growth-promoting activity was largely due to a greater supply of the essential amino acids. Some vitamin synthesis by the fungi is a possibility.

601

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MATERIALS AND METHODS

TABLE 1.—Composition of premix Ingredient

Percent

Glucose monohydrate Alfalfa meal (17% protein) Dicalcium phosphate Ground limestone Solka floe Salt mix' Vitamin mix 2 Methionine hydroxy analog Total

78.8 4.0 4.0 2.0 8.8 1.0 1.0 0.4 100.0

1

TABLE 2.—Composition of experimental rations (%) Dietary protein level Ingredient Premix Soybeans unextracted (properly processed) Glucose monohydrate

Protein, % M.E.,Kcal./Kg. M . E . / % P ratio Calcium, % Phosphorus, %

16% 50.0

17%

19%

50.0

15% 50.0

50.0

50.0

34.2 15.8

39.5 10.5

42.1 7.9

44.7 5.3

50.0 0.0

100.0

100.0

100.0

100.0

100.0

13%

13.34 3077 231 0.91 0.58

tory (Semeniuk et al., 1971) revealed that only 164 of 392 strains of Aspergilli were toxic. Some of the tested cultures exerted positive effects on growth of chicks and mice when the cultures were grown on sterile soybeans or wheat. The study reported here was designed to obtain further information on those cultures beneficial to chick growth and to attempt to provide explanations for these growthpromoting effects.

Calculated Analysis 16.34 15.34 17.34 3090 3085 3094 190 202 179 0.93 0.92 0.93 0.62 0.61 0.64

19.34 3102 161 0.95 0.67

soybeans were moistened, sterilized and airdried for each chick growth study for comparison with molded soybeans. The control or fermented soybeans were then incorporated into a premix (Table 1) in amounts to yield 15, 17, and 19% protein diets in Experi-

6. Aspergillus cultures used for four feeding experiments were obtained from the ARS Culture Collection, Northern Regional Research Laboratory, U.S.D.A., Peoria, Illinois.

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Containing 97% NaCl and other salts to provide 0.45% Mn, 0.50% Zn, 0.17% Fe, 0.05% Cu, 0.01% Co, 0.01% I and 0.30% S. Containing, per Kg., 1,000,0001.U. Vitamin A, 440,000 I.C.U. Vitamin D 3 , 4,400 I.U. Vitamin E, 220 mg. menadione, 360 mg. thiamine, 600 mg. pyridoxine, 0.36 mg. biotin, 880 mg. riboflavin, 1760 mg. pantothenic acid, 1.76 mg. cobalamine, 88 g. choline, 8.8 g. niacin and 22 g. ethoxyquin.

Inoculaof Aspergilli6 were grown on small amounts of soybeans (ca 25 g.) in 500 ml. Erlenmeyer flasks. To provide the amount of molded soybean needed (ca42 kg./culture for each experiment, good quality Chippewa soybeans were cracked, moisture conditioned to ca 31% moisture, sterilized (45 min. at 120° C.) and inoculated in plastic sacks as previously described (Semeniuk et al., 1970). The inoculated soybeans were then incubated in a chamber 2-3 days at 23-30° C , and shaken twice in the interim. Following incubation, which was just before sporulation had set in, the fermented soybeans were spread to air-dry 2-3 days on flat racks in a dry greenhouse hallway and finely ground in a coffee grinder. Uninoculated control

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C H A H , CARLSON, SEMENIUK, PALMER AND HESSELTINE

TABLE 3.—Essential amino acid content of control diets (%)'

TABLE 4.—Effect

of fermented soybeans on chick growth (Average 4-week weight)

Protein level

Control

15% 17% 19% mean 3

gm 477 555 540 524

Protein level 15% 17% 19% mean 3 Protein level 15% 17% 19% mean 3

Experiment V A. oryzae NRRL 2220

A. sydowi NRRL 5913

Mean

gm 485 571 527 528

gm 491 558 550 533

gm 512 548 582 547

Control 411 434 478 441

Experiment 2' A. clavatus A. oryzae NRRL 4 NRRL 696 460 451 453 482 485 488 463* 477**

A. oryzae NRRL 2220 442 467 484 464*

Mean 441 459 484 461

Control 418 457 512 462

Experiment 3' A . flavus A . flavus NRRL 450 NRRL 3518 517 496 539 518 571 552 535* 529*

A. sydowi NRRL 242 491 494 548 511*

Mean 481 502 546 509

Experiment 42 A. A. Protein A. oryzae A. oryzae candidus restrictus Mean level Control NRRL 451 NRRL 506 NRRL 1720 NRRL 147 451 455 462 473 13% 413 452 541 518 540 523 16% 492 523 587 582 19% 561 596 591 574 3 527* 519 524* 531** 522* mean 489 'AH values represent the average of 3 replicates of 10 male chicks each. 2 Each value represents the mean from 4 replicates of 7 male chicks. ' N o superscript indicates difference is not significant while * indicates significance at P < 0.05 and ** indicates significance at P < 0.01 as compared to the respective control diet.

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Chick Dietary protein level requirements 2 19% 17% 16% 15% 13% Amino acids (0-6 weeks) 0.98 Arginine 1.40 1.42 1.27 1.20 1.13 0.40 0.33 0.30 0.28 0.26 0.23 Cystine 0.46 0.46 0.41 0.38 0.36 0.31 Histidine 0.86 1.02 0.91 0.86 0.81 0.70 Isoleucine 1.60 1.42 1.27 1.20 1.13 0.98 Leucine 1.25 1.22 1.09 1.03 0.97 0.84 Lysine 0.46 0.27 0.24 0.22 0.21 0.18 Methionine 0.80 0.92 0.82 0.78 0.73 0.64 Phenylalanine 0.80 0.76 0.68 0.64 0.60 0.52 Threonine 0.23 0.28 0.25 0.23 0.22 0.19 Tryptophane 1.00 0.92 0.82 0.78 0.73 0.64 Valine 'In making calculations, the amino acid content of whole soybeans were used as given by Scott et al., (1969) Table 9.7, p. 438. 2 National Research Council, 1971.

FERMENTED SOYBEANS

603

TABLE 5.—Percentage growth responses for experimental groups Experiment 2 A . oryzae NRRL 696 iT9 11.1 2.1 8.4

A . sydowi NRRL 2220 T3 7.6 1.3 5.5

Mean 9?T 7.7 1.6 6.4

Protein level 15% 17% 19% mean

A. flavus NRRL 450 23?7 13.3 7.8 14.9

Experiment 3 A. flavus NRRL 3518 1877 17.9 11.5 16.0

A. sydowi NRRL 242 1775 8.1 7.0 10.9

Mean 20.0 13.1 8.7 13.9

Experiment 4 Protein level 13% 16% 19% mean

A . oryzae NRRL 451 9.4 6.3 6.2 7.3

A . oryzae NRRL 506 11.9 10.0 5.3 9.1

ments 1, 2, and 3, and 13, 16, and 19% protein diets in Experiment 4. Appropriate amounts of Cerelose were added to the lower protein rations as a replacement for the soybeans (Table 2). The experimental rations were formulated to be adequate with respect to the energy levels, minerals, and vitamins required for chicks as recommended by the N.R.C. (1971). The lower protein diets were deficient in amino acids (see Table 3). All feeding experiments were factorially arranged in a randomized complete-block design. Three replicates of 10 male chicks 7 each were used per treatment except in Experiment 4 where four replicates of 7 chicks were used. The chicks were housed in electrically heated battery brooders with raised wire floors to 4 weeks of age. Experimental diets and water were given ad libitum. Average chick weight gain and feed effi-

7. Golden Giant male broiler chicks, purchased from Jack Frost Hatcheries, Inc., St. Cloud, Minnesota.

A. candidus NRRL 1720 14.5 5.3 2.3 7.4

A. restrictus NRRL 147 10.2 9.8 4.6 8.2

Mean 11.5 7.9 4.6 8.0

ciency data were calculated. All mortalities were recorded. Percent dry matter digestibility and relative efficiency of nitrogen utilization were determined in Experiments 2 and 3 and proximate analyses of carcasses were made in Experiment 4. Samples of finely ground control and fermented soybeans were hydrolyzed by refluxing in 6 N HC1 under a nitrogen atmosphere at 110° C. for 24 hours. The hydrolyzates were analyzed for amino acids using a Beckman model 120C Amino Acid Analyzer. All data were subjected to analysis of variance as outlined by Steel and Torrie (1960). Where appropriate, Dunnett's procedure was applied to test for significant differences between each of the treatment means and the controls. RESULTS AND DISCUSSION Weight Gains. The average weight gain data of the four experiments are summarized in Table 4. In Experiment 1 only slight or inconsistent growth responses were obtained from the

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Protein level 15% 17% 19% mean

A . clavatus NRRL 4 9?7 4.4 1.5 5.2

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CHAH, CARLSON, SEMENIUK, PALMER AND HESSELTINE

TABLE 6.—Effect

Protein level 15% 17% 19% mean3

of fermented soybeans on feed efficiency (4-week) F/G ratios Experiment V A. oryzae NRRL 2220 1.75 1.65 1.55 1.65

Control 1.88 1.64 1.68 1.73

Protein level

Control

15% 17% 19% mean 3

1.87 1.85 1.72 1.81

Protein level

A. sydowi NRRL 5913 1.80 1.59 1.71 1.70

Experiment 2' A. clavatus A. oryzae NRRL 4 NRRL 696 1.77 1.75 1.66 1.73*

1.66 1.60 1.63 1.63*

Experiment 3' A. flavus A. flavus NRRL 450 NRRL 3518

Control

15% 17% 19%

0.05), indicating that the level of protein in the diet influenced the response obtained from different cultured soybeans. The percent improvement in chick growth due to each of the cultures tested with graded protein levels is given in Table 5. In Experiment 2, an average of all cultures showed the response to be 9.7%, 7.7%, and 1.6%, respectively, for the 15, 17, and 19% protein series. This compared favorably with values from Experiment 3 of 20.0%, 13.1%, and 8.7% for the same protein levels, and from Experiment 4 of 11.5%, 7.9%, and 4.6% for the 13, 16, and 19% protein series. This indicates that the chicks were indeed responding to an alteration of the protein composition of soybeans brought on by culturing. The great-

2.32 1.87 1.78 1.99

1.76 1.76 1.68 j 73**

1.82 1.71 1.68 1.74*

Experiment 42 Protein level

Control

A. oryzae NRRL 451

13% 16% 19% mean

2.01 1.98 1.88 1.96

1.96 1.79 1.73 1.83*

A. oryzae NRRL 506 1.83 1.76 1.66 1.75**

Mean 1.81 1.63 1.65 1.69

A. oryzae NRRL 2220

Mean

1.72 1.69 1.61 1.67*

1.76 1.72 1.66 1.77

A. sydowi NRRL 242

Mean

2.05 1.86 1.71 1.87*

1.99 1.80 1.71 1.83

A. Candidas NRRL 1720

A. restrictus NRRL 147

Mean

1.93 1.82 1.75 1.83*

2.04 1.80 1.68 1.84*

1.95 1.83 1.74 1.84

'Results are the average of triplicate groups of 10 chicks each. value represents the average of 4 replicates of 10 chicks each. No superscript indicates difference is not significant while * indicates significance at P < 0.05 and ** indicates significance at P < 0.01 as compared to the respective control diet. 2 Each 3

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treatments. The statistical analysis showed that neither the main effects nor any of their interactions were significant. However, there was an indication of a linear response to protein levels from the NRRL 2220 diet. With all other diets, maximum weight gains were obtained with 17% protein rather than 19%. In Experiments 2, 3, and 4 the data indicate superior growth for each of the fermented diets, as compared to the controls. In addition, there was good evidence for a greater enhancement of weight gains at the low protein levels. The analysis of variance revealed highly significant (P < 0.01) differences due to cultures and protein levels. The interaction of protein x culture in Experiment 3 was statistically significant (P <

605

FERMENTED SOYBEANS

est growth-promoting activities were shown with the lowest level of protein. Feed Efficiency. Data given in Table 6 show that cultured soybean diets gave significant improvements (P < 0.05 and < 0.01) in feed utilization in Experiments 2, 3, and 4 but not Experiment 1. Since the interactions of protein x culture in feed efficiency were not significant in any of the experiments, it is concluded that the responses were due to nutritional qualities of the cultures. Mortality was of no consequence

TABLE 7.—Effect

Protein level

of fermented soybeans on relative efficiency of nitrogen utilization (%)'

Control

%

15% 17% 19% nean

20.23 26.20 28.46 24.96

Protein level

Control

15% 17% 19% mean

30.80 28.26 23.13 27.40

Experiment 2 A. clavatus A. oryzae NRRL 4 NRRL 696

%

23.23 29.69 28.10 27.01

%

34.65 26.00 27.61 29.42

Experiment 3 A. flavus A. flavus NRRL 450 NRRL 3518 28.76 33.13 25.52 29.14

29.84 26.73 26.78 27.78

A. oryzae NRRL 2220

%

Protein level

Control

15% 17% 19% mean

52.56 50.35 49.45 50.79

Protein level

Control

15% 17% 19% mean

66.97 62.86 58.53 62.79

%

25.25 30.80 30.77 28.94

25.84 28.17 28.74 27.58

A. sydowi NRRL 242

Mean

24.86 25.29 25.42 25.19

28.57 28.35 25.21 27.38

'Nitrogen efficiency was measured during the last 3 days of each experiment. TABLE 8.—Effect

Mean

N-retention N-intake

x 100.

of fermented soybeans on dry matter digestibility (%)' Experiment 2 A. clavatus A. oryzae NRRL 4 NRRL 696 54.35 55.76 49.33 53.15

58.22 56.65 55.31 56.73

Experiment 3 A. flavus A. flavus NRRL 450 NRRL 3518 68.33 69.35 62.42 66.70

68.23 63.69 62.92 64.95

A. oryzae NRRL 2220

Mean

59.60 58.45 57.73 58.59

56.18 55.30 52.96 54.81

A. sydowi NRRL 242

Mean

65.00 64.16 61.65 63.60

67.13 65.02 61.38 64.51

'Dry matter digestibility was determined during the last 3 days of the 28-day experimental period. (Wt. dry feed consumed—Wt. dry excreta) x 100.

Wt. dry feed consumed

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Mortality.

throughout Experiments 1,2 and 4. In Experiment 3 considerable mortality was encountered among the control birds. The final mortality data showed losses of 36.7%, 6.7% and 3.3% for the 15, 17 and 19% protein series, respectively. About a week after the initiation of the experiment, several chicks in the control 15% protein diet exhibited a somewhat typical polyneuritis characterized by a "star-gazing" position. Upon injecting 2-3 mg. thiamine, they quickly recovered. Therefore, all diets were immediately supplemented with twice the usual level of thiamine and this level was used in Experiment 4. The

606

CHAH, CARLSON, SEMENIUK, PALMER AND HESSELTINE

lack of polyneuritis symptoms in chicks fed cultured soybeans indicated that the cultures were presumably synthesizing thiamine. A further study would be necessary to explore this possibility. Production of vitamins by the fungi could account for some of the growth effects.

TABLE 9.—Carcass proximal analysis data (%) Protein level

Control

A. oryzae NRRL 451

13% 16% 19% mean 7

58.25' 60.33 59.83 59.47"

57.50 57.67 58.00 57.72"

A. oryzae NRRL 506 Moisture2 56.34 56.67 58.00 57.00"

A. Candidas NRRL 1720

A. restrictus NRRL 147

Mean

57.17 57.33 57.67 57.39"

61.33 60.25 60.34 60.64"

58.12 58.45 58.77 58.45

43.74 42.64 42.86 43.08"

44.78 43.90 38.15 42.28"

45.65 41.82 40.42 42.63

13% 16% 19% mean 7

47.15 44.50 45.99 45.88"

43.75 37.99 34.29 38.68"

Fat3-4 43.81 40.08 40.79 41.56 b

13% 16% 19% mean 7

41.18 43.15 45.85 43.39"

42.53 45.29 46.48 44.77"

Protein*5 44.27 47.23 47.66 46.39"

44.57 47.61 48.12 46.77"

46.50 47.40 48.26 47.39"

43.81 46.14 47.27 45.52

13% 16% 19% mean 7

6.39 7.22 7.89 7.17"

8.23 8.80 8.71 8.58"

Ash36 8.71 8.67 8.81 8.73"

8.34 8.50 8.00 8.28"

7.98 8.11 8.28 8.12"

7.93 8.26 8.34 8.18

'Each value represents the average of duplicate samples per replicate.

2 Moisture (% of total carcass)—based on water loss upon lyophilization. 3 Fat, Protein and Ash were determined based on % of dry weight. 4 Fat by a 12 hour Soxhlet extraction with diethyl ether. 5 Protein (N x 6.25) by the Kjeldahl method. 6 Ash after a 2 hour at 600° C. incineration. 7

Within each parameter, means not having common letter superscripts were significantly different from the respective control at the 5% level of probability.

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Nitrogen Efficiency and Apparent Digestibility. Nitrogen dry matter utilizations were measured in Experiments 2 and 3. Nitrogen efficiency data are presented in Table 7. Dry matter digestibility data are presented in Table 8. It should be pointed out that these data were obtained during the last 3 days of the 28-day experimental period. The

nitrogen efficiency data show that the chicks responded to something in the culture other than just increased protein levels. This was especially evident in Experiment 3 where the control chicks on the lower protein diets had higher nitrogen utilization values than the chicks fed cultured soybeans. Except for this instance, chicks that received cultured soybeans utilized dietary nitrogen more effectively than birds fed control diets. Dry matter digestibility was somewhat consistently superior for the chicks fed cultured soybeans, especially in Experiment 2. These two sets of data suggest that some substantial changes in availability of nutrients had occurred in the cultured soybeans.

607

FERMENTED SOYBEANS

TABLE 10.—Essential amino acids in fermented soybeans' (m moles/100 g. soybean) Experiment 1 Arginine Histidine Isoleucine Leucine Lysine Methionine Phenylalanine Threonine Valine Glycine

Amino acid Arginine Histidine Isoleucine Leucine Lysine Methionine Phenylalanine Threonine Valine Glycine

Amino acid Arginine Histidine Isoleucine Leucine Lysine Methionine Phenylalanine Threonine Valine Glycine

Control 16.5 8.0 15.5 28.5 14.0 4.0 12.5 15.0 17.5 25.0

Control 19.5 8.5 14.0 27.0 18.5 4.5 14.5 15.5 16.0 29.0

A. oryzae NRRL 2220 17.5 8.0 15.5 29.0 20.0 4.5 15.5 16.5 18.5 27.0

Experiment 2 A. clavatus NRRL 4 19.0 6.5 13.5 24.5 18.0 5.0 15.0 18.0 19.5 31.0 Experiment 3 AL. flavus NRRL 45C1 20.5 7.0 17.5 31.5 21.0 5.0 17.5 18.0 20.5 29.5

A. sydowi NRRL 5913 18.0 8.0 17.0 31.5 21.0 5.0 17.0 18.0 20.0 29.5

18.5 10.5 18.5 30.0 19.5 4.0 17.5 17.5 21.5 28.5

A. oryzae NRRL 2220 20.5 8.5 17.0 31.5 19.0 5.0 15.5 18.0 19.5 29.5

A. flavus NRRL 3518 20.5 9.0 15.0 27.0 20.5 4.5 18.0 15.5 17.5 33.0

A. sydowi NRRL 242 17.0 6.0 17.0 29.5 17.0 5.0 17.0 17.5 20.0 29.0

A. oryzae NRRL 696

Experiment 4 A. oryzae Amino acid Control NRRL 451 Arginine 18.10 20.85 Histidine 10.70 10.65 Isoleucine 18.30 20.70 Leucine 32.30 35.05 Lysine 19.80 22.75 Methionine 4.20 4.35 Phenylalanine 15.45 17.60 Threonine 16.75 20.04 Valine 18.20 21.70 Glycine 27.80 32.55 'Measured by Beckman 120 C Auto Analyzer.

A.. oryzae NRRL 506 21.80 11.70 18.85 42.35 21.55 3.86 16.65 18.80 25.65 31.50

A. candidus NRRL 1720 20.15 13.30 20.95 33.40 21.05 4.95 16.50 18.15 23.45 30.10

A. restrictus NRRL 147 21.40 9.85 17.35 29.55 21.95 4.45 15.60 17.25 20.45 28.45

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Control 16.0 8.5 16.0 23.5 18.5 4.5 15.0 16.5 17.0 31.0

Amino acid

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CHAH, CARLSON, SEMENIUK, PALMER AND HESSELTINE

assistance in the form of a contract, No. 12-14-100-11017(71), from the United States Department of Agriculture to aid in conducting these investigations. Thanks are also extended to Dr. W. F. Kwolek, statistician at the Northern Regional Research Laboratory, U.S.D.A., Peoria, Illinois, and to Dr. W. L. Tucker, Agricultural Experiment Station Statistician, S.D.S.U., for their assistance with statistical analyses of the data.

Amino Acid Composition. The essential amino acid composition data are shown in Table 10. The data indicate that cultured soybean preparations were consistently higher than the controls in several essential amino acids, particularly arginine, lysine, threonine and valine. It is well known that the nutritional value of a protein is determined primarily by its amino acid balance, both quantitatively and qualitatively (Harper, 1965; Sugahara et al., 1969). For example, Cromwell et al. (1968) reported that chick diets containing more adequate amounts of lysine and methionine produced faster and more efficient weight gains than the controls. In view of the nutritional significance of amino acid combinations and of limiting amino acids in the efficiency of protein utilization, the additional amounts of essential amino acids in the cultured soybeans are considered to be largely responsible for the superior growth and more efficient feed conversion they promoted. A further study is needed to establish these effects. Amino acids should be added to a control soybean diet to simulate the amino acid profiles of the cultured soybeans. The fact that the greatest growth responses were obtained with the lowest protein levels points to an amino acid effect.

Allcroft, R., 1969. Aflatoxicosis in farm animals. In: Aflatoxin, Goldblatt, L. A., ed., Academic Press, New York and London. Blount, W. P., 1961. Turkey " X " disease. Turkeys (J. Brit. Turkey Federation), 9: 52-61. Choudhury, H., C. W. Carlson and G. Semeniuk, 1971. A study of ochratoxin toxicity in hens. Poultry Sci. 50: 1855-59. Chute, H. L., S. L. Hollander, E. S. Barden and D. C. O'Meara, 1965. The pathology of mycotoxicosis of certain fungi in chickens. Avian Dis. 9: 57-66. Cromwell, G. L., J. C. Rogler, W. R. Featherston and T. R. Cline, 1968. A comparison of the nutritive value of opaque-2, floury 2 and normal corn for the chick. Poultry Sci. 47: 840-847. Doupnik, B., Jr., and J. C. Peckham, 1970. Mycotoxicity of Aspergillus ochraceus to chicks. Appl. Microbiol. 19: 594-597. Forgacs, J., 1966. Mycoses and mycotoxicosis in poultry. Part II. Feedstuffs, 38(11): 26, 30, 71. Garlich, J. D., H. T. Tung and P. B. Hamilton, 1972. Delayed effects of aflatoxin on egg production. Poultry Sci. 51: 1870. Harper, A. E., 1965. Effect of variations in protein intake on enzymes of amino acid metabolism. Can. J. Biochem. 43: 1589-1603. Hamilton, P. B., and J. D. Garlich, 1971. Aflatoxin as possible cause of fatty liver syndrome in laying hens. Poultry Sci. 50: 800-804. Meronuck, R. A., K. H. Garren, C. M. Christensen, G. H. Nelson and F. Bates, 1970. Effects on turkey poults and chicks of rations containing corn invaded by Penicillium and Fusarium species. Amer. J. Vet. Res. 31: 551-555. National Research Council, 1971. No. 1, Nutrient Requirements of Poultry, ISBNO-309-01861-7., National Academy of Sciences, Washington, D.C. Richardson, L. R., S. Wilkes, J. Godwin and K. R. Pierce, 1962. Effect of moldy diet and moldy soy-

ACKNOWLEDGEMENT The authors wish to acknowledge financial

REFERENCES

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Carcass Analyses. Table 9 shows carcass analysis data for Experiment 4. The data were obtained by analyzing the total carcasses of two chicks, the heaviest and lightest, from each replicate within treatment upon termination of the experiment. The results indicate that diets made with cultured soybeans produced chicks with significantly higher (P < 0.05) protein and ash content and significantly lower (P < 0.05) total lipids than the control diets produced. Chicks receiving cultured soybeans thus appeared to produce a carcass of higher nutritional value.

FERMENTED SOYBEANS

1970. A study of aflatoxicosis in laying hens. Poultry Sci. 49: 1082-84. Smith, J. W., and P. B. Hamilton, 1970. Aflatoxicosis in the broiler chicken. Poultry Sci. 49: 207-215. Speers, G. M., R. A. Meronuck, D. M. Barnes and C. J. Mifocha, 1971. Effect of feeding Fusarium roseum F. sp. graminearum contaminated corn and the mycotoxin F-2 on the growing chick and laying hen. Poultry Sci. 50: 627-633. Steel, R. D. G., and J. H. Torrie, 1960. Principles and Procedures of Statistics. McGraw-Hill Book Co., New York. Sugahara, M., D. H. Baker and H. M. Scott, 1969. Effect of different patterns of excess amino acids on performance of chicks fed amino acid deficient diets. J. Nutr. 97: 29-32. Tucker, T. L., and P. B. Hamilton, 1971. The effect of ochratoxin in broilers. Poultry Sci. 50: 1637. Wyatt, R. D., and P. B. Hamilton, 1972. The effect of rubratoxin in broiler chickens. Poultry Sci. 51: 1383-87.

Use of Dried Poultry Waste in Diets for Chickensl N . TRAKULCHANG AND S. L . BALLOUN

Department of Animal Science, Iowa State University, Ames, Iowa 50010 (Received for publication July 31, 1974)

ABSTRACT Two experiments with broiler chicks and one with laying hens were conducted to investigate the effects of dried poultry feces (DPW) in diets for poultry. Supplementing 10% DPW, or 20% DPW with added amino acids, in a low-protein (15% crude protein) practical diet did not affect weight gains and feed efficiency significantly (P < 0.01). Nitrogen utilization and nitrogen gain of broilers were significantly decreased (P < 0.01) by DPW. When laying hens were fed diets containing DPW, egg production rate and feed efficiency were decreased, and mortality was increased (P < 0.05). POULTRY SCIENCE 54: 609-614, 1975

INTRODUCTION LEGAL and Zindel (1970a) found that weight gain and feed efficiency of chicks were inversely related to the levels of DPW in the diet. On the contrary, Lee and Blair (1972, 1973) reported that chicks fed a crystalline amino acid diet plus dried poultry manure gained more than those fed the diet plus added essential amino acids.

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1. Journal Paper No. J-7968 of the Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa. Projects 1932 and 1984.

York et al. (1970) reported that egg production, shell thickness and egg weight were not affected by DPW in the diets but that feed efficiency was inversely proportional to the amount of DPW in the diet. Blair and Lee (1973) reported improvements of egg production, feed efficiency and body weight when 9.7% dried, autoclaved poultry manure was added to a low-protein layer diet. In contrast, Flegal and Zindel (1970b) showed that egg production decreased as the levels of DPW in the diets increased. The following experiments were conducted

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bean meal on the growth of chicks and poults. J. Nutr. 78: 301-306. Scott, M. L., M. C. Nesheim and R. J. Young, 1969. Nutrition of the Chicken. M. L. Scott and Assoc, Ithaca, N.Y. Scott, P. M., and E. Somers, 1969. Biologically active compounds from field fungi. J. Agri. Food Chem. 17: 430-436. Semeniuk, G., G. S. Harshfield, C. W. Carlson, C. W. Hesseltine and W. E. Kwolek, 1970. Occurrence of mycotoxins in Aspergillus, pp. 185-190. Proc. First U.S.-Japan Conf. Toxic Microorg., U.S. Govt. Printing Office, Washington, D.C. Semeniuk, G., G. S. Harshfield, C. W. Carlson, C. W. Hesseltine and W. F. Kwolek, 1971. Mycotoxins in Aspergillus. Mycopath. Mycol. Appl.43: 137-152. Sharby, T. F., G. E. Templeton, J. N. Beasley and E. L. Stephenson, 1973. Toxicity resulting from feeding experimentally molded corn to broiler chicks. Poultry Sci. 52: 1007-1014. Sims, W. M., Jr., D. C. Kelley and P. E. Sanford,

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