NUMEROUS investigations have

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1 Use of Equivalent Levels of Antibiotic and Vitamin B Supplements With Cottonseed and Soybean Oil Meals in Chick Diets 1 NUMEROUS investigations have been conducted on the growth responses produced by soybean oil meal diets supplemented with antibiotics and vitamin Bi 2. In contrast, comparatively few such studies have been conducted with cottonseed meal as a protein source in all-vegetable rations. One such study reported by Sherwood and Couch (19) indicates that antibiotics and vitamin B i2 possess growth promoting properties when used as supplements to cottonseed meal diets. These workers supplemented a milo-cottonseed diet with a 1 percent APF concentrate (aureomycin fermentation product) and obtained growth superior to that produced by an unsupplemented basal diet. Richardson and Blaylock (19) reported that cottonseed meal supplemented with lysine and vitamin B12 supported a rate of growth comparable to that obtained when soybean meal was used as a source of protein in a chick ration, but resulted in inferior rate of growth when vitamin B12 and lysine were omitted. Machlin, Denton and Bird (192) obtained an increase in growth by supplementing a corn-cottonseed meal basal diet with an aureomycin fermentation product containing B12 and aureomycin. They also reported that the addition of this product to a lysine-supplemented basal diet produced growth superior to that obtained from a good commercial 1 Contribution No. 2, Department of Poultry- Husbandry. R. W. LEWIS AND P. E. SANFORD Kansas State College, Manhattan, Kansas (Received for publication August 9, 192) 268 broiler ration. These investigations suggest that further studies regarding antibiotic and vitamin B12 supplementation of cottonseed meal diets, might make possible improvement of the nutritional value of the diets to the extent that such a source of vegetable protein in addition to soybean meal may be efficiently utilized in practical chicks diets. The present study was conducted to compare the growth responses promoted by supplementing soybean and cottonseed meal diets with equivalent levels of various commercial antibiotic and vitamin B12 feeding supplements. The extent and conditions under which cottonseed meal can be used in practical chick diets was also studied. MATERIALS AND METHODS Chicks used were an Fi cross of New Hampshire X Kansas State College Strain White Plymouth Rocks. All chicks were reared on wire-floored batteries. Two hundred chicks were randomized into 1 lots of 2 chicks each for the first experiment. Sexed chicks were used in the second experiment, with 11 males and 1 females in each of 1 lots. The birds were weighed at one day of age and every seven days thereafter throughout the duration of the two experiments. Room temperature was controlled by thermostat, regulated between 7 and 7 degrees Fahrenheit, and heated by natural gas. The following antibiotic and vitamin Downloaded from at Penn State University (Paterno Lib) on May 9, 216

2 ANTIBIOTICS AND B i2 IN PLANT PKOTEIN DIETS 269 B12 feeding supplements were used: Aurofac, containing 1.8 grams of aureomycin fermentation product and 1.8 milligrams of B12 per pound of supplement (7.6 grams per 1 pounds of diet); crystalline pure aureomycin HCl (21 milligrams per 1 pounds of diet); Bacitracin supplement, containing 3 grams of bacitracin fermentation product and 3 milligrams of B12 per pound of supplement (4.4 grams per 1 pounds of diet); Bacitracin supplement containing grams of bacitracin fermentation product per pound (18.2 grams per 1 pounds of diet); Bi-Con- 3+3 containing 3 grams of terramycin fermentation product and 3 milligrams of B12 per pound (4.4 grams per 1 pounds of diet); and TM- containing grams of terramycin fermentation product per pound (18.2 grams per 1 pounds of diet). The amount of different supplements was adjusted so as to add a level of 1 grams of antibiotic and 6 milligrams of vitamin B12 per ton of feed. Two basal diets were used in the first experiment. Basal 1 was that used by the Kansas Agricultural Experiment Station and referred to as the Kansas State College high efficiency, all-vegetable protein ration. Antibiotic and vitamin B12 feeding supplements were omitted. Basal 2 was modified from the Kansas State College diet by substituting 33 percent of a commercial hydraulic cottonseed meal (containing 41 percent protein) for the soybean meal. The vitamin, mineral and amino acid content of Basal 2 was calculated and compared with the National Research Council's Recommended Nutrient Allowance for Poultry (19). The composition of the diets is given in Table 1. As it was recognized that cottonseed meal is deficient in lysine, all diets containing cottonseed meal in the first experiment were supplemented with.2% L-lysine. The concentration of the antibiotics and vitamin B12 supplements was calculated for each product and adjusted so that each was supplemented at the previously mentioned level of 1 grams of antibiotic and 6 milligrams of B12 per ton of feed. This level was chosen as a practical and optimum level of supplementation as a result of experiments conducted by Sanford (192a) at the Kansas Agricultural Experimental Station. The concentration of antibiotics and B12 supplements was not sufficient to attain a level of 1 grams when used alone, thus it was necessary to include crystalline aureomycin HCl (2 milligrams per 1 pounds of diet); Bacitracin ( grams of bacitracin per pound of supplement 18.2 grams per 1 pounds of diet); Bi-Con TM- ( grams of terramycin per pound of supplement 18.2 grams per 1 pounds of diet), to attain this level. Diets la, 2a, 3,4, and 6 were fed in the second experiment (Table 2). They were prepared three times during each 8-week experimental feeding period. In the second experiment, the cottonseed meal included in diets 3, 4, and 6 was an improved screw-pressed type, experimentally processed under the following specific conditions: maximum temperature in cooker, 2 degrees F.; moisture content of meats leaving cooker 4%; gossypol content,.3%. The commercial hydraulic type meal used in both experiments was processed as follows: maximum temperature in cooker, 23 degrees F.; moisture content of meats leaving cooker, 8.4%, gossypol content, 2.%. Upon the recommendation of workers at the USDA Southern Regional Research Laboratory, L-lysine (monohydrochloride, anhydrous) 2 We are indebted to Dr. F. H. Thurber, Chemist, Protein and Carbohydrate Division, USDA Southem Regional Research Laboratory, for the gossypol anaylsis of the hydraulic type cottonseed meal. Downloaded from at Penn State University (Paterno Lib) on May 9, 216

3 27 R. W. LEWIS AND P. E. SANFORD TABLE 1.Composition of the basal diets for Experiment 1 Ingredients Ground yellow corn Wheat bran Alfalfa meal (17% dehyd.) Soybean oil meal (44% solvent ext.) Cottonseed oil meal (41% hydraulic) Calcium carbonate Steamed bone meal Salt (NaCl) Total Manganese sulfate "Prot A" 1 "Delsterol"D 2 Riboflavin Choline chloride (crystalline) Calcium pantothenate Niacin Lysine HC1 (anhydrous) Antibiotics and B12 feeding supplements per 1 lb. 3 "Aurofac" (1.8 g. aureomycin+1.8 mg. B12 per lb. of supplement) Aureomycin HC1 (crystalline) "Bacitracin 3-3" (3 g. bacitracin+3 mg. B12 per lb. of supplement) "Bacitracin " ( g. bacitracin per lb. of supplement) "Bi-Con 3+3" (3 g. terramycin+3 mg. B12 per lb of supplement) "TM-" ( g. terramycin per lb. of supplement) 1 Supplies 2,4 USP units of vitamin A per g. of supplement. 2 Supplies 2, A.O.A.C. units of vitamin D3 per g. of supplement. 3 Supplemented at the rate of 1 g. of antibiotics and 6 mg. of B12 per ton. was added at a level of 4.2 grams per pound of cottonseed meal used in the second experiment. In view of findings in the first experiment, a combination antibiotic supplement consisting of Aurofac and Bacitracin was added to the basals in the second experiment at the same level. RESULTS AND DISCUSSION Experiment 1. The results presented in Table 3 show that the growth of chicks fed the cottonseed meal diets was inferior in all cases to that of chicks receiving the soybean meal diets. An analysis of variance of the weights at 8 weeks revealed highly significant differences in the growth of the 1 lots of chicks. This supports the findings of Richardson and Blaylock (19) and Sherwood and Couch (19) 1 6 lb Basal diet 2 8.1b grams per 1 pounds 2. g.. g g. 2 mg. 4.4 g g. 4.4 g g. who reported that diets containing cottonseed meal as the source of protein in an all-vegetable ration produced growth inferior to that attained by feeding soybean meal diets when neither was supplemented with vitamin B12 and lysine or antibiotics. Observations in the present study reveal that although chicks fed unsupplemented cottonseed meal basal diets (Lot 2) were inferior in growth, and grew at a slower rate, the birds were more uniform in size than those receiving the unsupplemented soybean meal basal diet (Lot 1). There were highly significant differences between the growth of chicks in lots receiving antibiotics and vitamin B12 and that of chicks receiving the nonsupplemented diets. The birds fed the cottonseed meal diets supplemented with the Aurof ac-bacitracin combination Downloaded from at Penn State University (Paterno Lib) on May 9, 216

4 ANTIBIOTICS AND B12 IN PLANT PROTEIN DIETS 271 TABLE 2.Composition of the basal diets for Experiment 2 TlmfO/lionto itigrcuiciila Ground yellow corn Wheat bran Alfalfa meal (17% dehyd.) Soybean oil meal (44% solvent ext.) Cottonseed meal (41% screw-pressed) Cottonseed meal (41% hydraulic) Meat scraps (% protein) Fishmeal (6% protein) Calcium carbonate Steamed bone meal Salt (NaCl) Total Manganese sulfate "Prot A" 1 2 "Delsterol"D 3 Riboflavin Choline chloride (crystalline) Calcium pantothenate Niacin Lysine HC1 (anhydrous) Antibiotic and B12 feeding supplement _ la 6 1b ' g per 1 lb. 1 "Aurofac" (1.8 g. aureomycin+1.8 mg. B12 per lb. of supplement) Aureomycin HC1 (crystalline) Bacitracin (3 g. bacitracin-)-3 mg. B : 12 per lb. of supplement) Bacitracin ( g. bacitracin per lb. of supplemen t) i. 2a 8. lb g lb Diets grams per 1 pounds. g.. g Supplies, USP units of vitamin A per g. of supplement. 2 Supplies 2, A.O.A.C. units of vitamin D 3 per g. of supplement 3 Supplemented at the rate of 1 g. of antibiotics and 6 mg. of Bis per ton. maintained an average weight of grams more than those fed the unsupplemented diet (Lots 2 and 1, Table 3). This compares with an increase of 1 grams in weight reported by Sherwood and Couch (19) as a result of supplementing a milo-cottonseed meal diet with a 1 percent APF concentrate. The cottonseed meal in that study was fed at a level of 3% of the diet and contained.3% gossypol. The increase in mean weight due to supplementation of the soybean meal diet with Aurofac-Bacitracin " combination was grams (Lots 1 and 9, Table 3). Under the conditions of the present study it was found that few significant growth differences between lots resulted from the addition of different antibiotic and vitamin B12 supplements to either the 4 9. lb g. mg g. 9.1 g. 66. lb g b g soybean or the cottonseed meal basal diet. This result coincides to a degree with the findings of Matterson el al. (191) regarding their work with antibiotics and soybean all-vegetable protein diets. The investigations herein reported show that the combination of the antibiotic supplements Aurofac and Bacitracin appeared to be the most consistent of all the supplements used, with respect to growthpromoting properties when added to the cottonseed meal diet. Davis and Briggs (191) combined aureomycin HC1 with streptomycin and also procaine penicillin G with bacitracin. These authors did not report any evidence of superior growth over that produced by single antibiotics used to supplement a soybean meal diet. There is a slight possibility that the combination observed in Downloaded from at Penn State University (Paterno Lib) on May 9, 216

5 272 R. W. LEWIS AND P. E. SANFORD TABLE 3.Average weight, mortality, and feed efficiency of chicks to eight weeks for Experiment 1 Lot : Gm. feed per gm. gain. Basal 1 Basal 2 Basall+Aurofac Basal 2+Aurofac Basal 1+Bacitracin Basal 2+Bacitracin Basal 1+Terramycin Basal 2+Terramycin Basal 1+Aurofac-Bacitracin Basal 2+Aurofac-Bacitracin this study may be better adapted to cottonseed meal diets when used as a mixture. Work by Sanford (192b) more adequately treats the synergistic effect of antibiotic and vitamin B 12 feeding supplements fed with soybean meal diets. The investigations cited were designed primarily to obtain information as to the comparative growth-promoting effects of these combinations. Growth was too variable within the lots, and responses varied too greatly with regard to sex for a definite sequential evaluation of the supplements discussed in the present report. Mean weight in grams , , Mortality, % Feed efficiency* The feed efficiency of both basal diets was considerably improved when antibiotics and vitamin B12 were added (Table 3). This result is in agreement with the findings of Davis and Briggs (191) and Matterson et al. (191). These authors reported improved feed efficiency when soybean meal diets were supplemented with antibiotics. Mortality was negligible during the experimental period, as indicated in Table 3. The appearance of the birds receiving the soybean meal basal was superior to that of those fed the cottonseed meal basal. Between the sixth and eighth week, the chicks fed the nonsupplemented cottonseed meal diet seemed to "tire" of the diet. Weekly feed consumption remained constant, in contrast to an increased consumption of feed by the chicks receiving the nonsupplemented Basal 1 (soybean) and those receiving either cottonseed meal or soybean meal supplemented with antibiotics and vitamin B12. In the eighth week several birds showed symptoms characteristic of riboflavin deficiency. The condition was found in lots receiving the cottonseed meal diets, irrespective of supplementation. Grau (19) reported such a condition in his investigation with cottonseed meal when the product was included at levels above 3% in chick diets. Experiment 2. The changes in diets for Experiment 2 consisted primarily of the following: vitamin D supplement reduced to 2 grams (to conform with the recent standard International Chick Unit, 7 I.C.U.'s equivalent to 1 A.O.A.C. units); vitamin A supplement decreased to 4 grams, compared with 1 grams used in the first experiment (due to an increase to, U.S.P. units per gram in the potency of the new product used in the second experiment); riboflavin content increased from 1.76 to 3.2 milligrams per pound of diet because of the "curled toe" condition noted in the first experiment. It was observed in the second experiment that the diet containing screwpressed cottonseed meal supplemented Downloaded from at Penn State University (Paterno Lib) on May 9, 216

6 ANTIBIOTICS AND BI 2 IN PLANT PROTEIN DIETS 273 TABLE 4.Average weight, mortality and feed efficiency of chicks to eight weeks for Experiment 2 l Lot Diet la 2af 3t la+aurofac-bacitracin 2af+Aurofac-Bacitracin 3J+Aurofac-Bacitracin 4J+12% Soybean Meal+Aurofac- Bacitracin 4J+12% Soybean Meal+Aurofac-Bacitracin +.187% lysine t+6% Meat Scraps+6.S% Fish Meal+ Aurofac-Bacitracin 6f+2% Meat Scraps+3% Fish Meal+1% Soybean Meal+Aurofac-Bacitracin Mean weight in grams ± ± OS Mortality, % Feed efficiency* * Gm. feed per gm. gain. f Diets containing hydraulic type cottonseed meal. I Diets containing screw-pressed type cottonseed meal. 1 The mean weights, feed efficiencies, and percent of mortality in Experiment 2 do not compare favorably with those recorded in Table 3, Experiment 1, due to the occurrence of a respiratory disease in alllots during the fifth week of the second experiment. with antibiotics and vitamin Bj2 resulted in superior growth when compared with the diet containing the hydraulic cottonseed meal supplemented in the same manner (Table 4). The difference in the mean weights of the birds fed these two diets was grams, a highly significant difference. When the types of meal were compared without supplementation, no significant difference was noted between the mean weights of the birds receiving the screw-pressed cottonseed and those of the birds receiving hydraulic type meal. Inclusion of the Aurofac- Bacitracin combination, 6% meat and bone scraps, and 6.% fish meal in the screw-pressed cottonseed meal diet produced an increase of 37.9 grams. Diet 6, containing the combination of Aurofac and Bacitracin, 1% screw-pressed cottonseed meal, 2% meat and bone scraps, 3% fish meal, and 1% soybean meal produced growth comparable to that produced by Diet la supplemented with Aurofac-Bacitracin (Kansas State College high efficiency broiler ration). These findings apparently confirm the report by Sherwood and Couch (19) that cottonseed meal can be incorporated in the diet of chicks and promote satisfactory growth when properly supplemented. Addition of.187% lysine to the diet containing 12% soybean meal, 2% cottonseed meal, and the combination of Aurofac and Bacitracin supplements did not promote a significant further increase in growth rate. Further observations indicated that the screw-pressed meal diet supplemented with 6% meat and bone scraps, 6.% fish meal, and the Aurofac- Bacitracin combination produced growth significantly greater than that promoted by the all-vegetable protein 2% screwpressed cottonseed meal diet containing 12% soybean meal, the Aurofac-Bacitracin combination and.187% lysine. These observations support the findings of Milligan et al. (191) whose work indicated that lysine could be omitted from a cottonseed meal diet and satisfactory growth attained, provided an adequate source of animal protein is included. Grau (19) reported similar results. The feed efficiencies of all supplemented Downloaded from at Penn State University (Paterno Lib) on May 9, 216

7 274 R. W. LEWIS AND P. E. SANFORD screw-pressed meal diets were more favorable than that of the supplemented diet containing hydraulic type meal. As there were no symptoms characteristic of riboflavin deficiency as noted in the first experiment, it is assumed that Grau's (19) hypothesis regarding the higher riboflavin requirement of chicks fed high levels of cottonseed meal was confirmed. In the fifth week of the experiment, the occurrence of a respiratory disease markedly retarded the growth, and altered feed efficiencies of birds in all lots. Mortality was also increased. A comparison of Tables 3 and 4 for both experiments, however, will show only a slight difference in the eight week weights of the birds recorded in both experiments. After recovery, the birds grew satisfactorily and it is believed that the results obtained due to the disease outbreak did not seriously hinder or alter the pattern of the experiment, as all lots were exposed simultaneously. There was no significant change in the growth pattern after recovery from the disease. SUMMARY The findings of this study may be summarized as follows: 1. Antibiotic and vitamin B12 feeding supplements promote growth in chicks when added to a cottonseed meal allvegetable protein diet. 2. The response promoted by adding antibiotic and vitamin B12 supplements to a cottonseed meal all-vegetable protein diet was inferior to that obtained in chicks fed soybean meal diets supplemented in an identical manner. 3. The combination of the antibiotics' supplements Aurofac and Bacitracin appeared to be more consistent in growthpromoting properties than other supplements to a cottonseed meal all-vegetable protein diet. 4. The experimentally processed screw-pressed meal promoted growth superior to that produced by the hydraulic type meal when both were supplemented with the Aurofac-Bacitracin combination.. The incorporation of 12% soybean meal in the all-vegetable protein diet containing 2% screw-pressed cottonseed meal and a combination of Aurofac and Bacitracin apparently compensated for the lysine deficiency in cottonseed meal. 6. Screw-pressed cottonseed meal included at a level of 1% in a practical ration adequately supplemented with a source of animal protein, soybean meal, antibiotics and vitamin B12, compared favorably to the Kansas State College high efficiency ration with respect to resulting growth rate of chicks. ACKNOWLEDGEMENT The authors express their appreciation to the following: Dr. A. M. Altschul of the U.S.D.A. Southern Regional Research Laboratory, New Orleans, La. for the samples of experimentally processed screw-pressed cottonseed meal; Lederle Laboratories, American Cyanamid Co., Pearl River, N. Y. for the Aurofac and aureomycin HC1; U. S. Industrial Chemicals, Inc., New York, N. Y. for the Bacitracin 3 3 and Bacitracin 1113B; Chas. Pfizer and Co. Inc., Brooklyn, N. Y. for the Bi-Con 3+3 and TM-, and Merck and Co., Rahway, New Jersey for the lysine. REFERENCES Cravens, W. W., H. J. Almquist, R. M. Bethke, H. R. Bird and L. C. Norris, 19. Recommended nutrient allowances for poultry. National Research Council, Washington, D. C. Davis, R. L., and G. M. Briggs, 191. Studies with antibiotics in chick and poult starting rations. Poultry Sci. 3: Grau, C. R., 19. Cottonseed meal for chicks. Cali- Downloaded from at Penn State University (Paterno Lib) on May 9, 216

8 HOME PRESERVATION AND EGG QUALITY 27 fornia Agriculture 4 (No. 9): Machlin, L. J., C, A. Denton and H. R. Bird, 192. Supplementation with vitamin B12 and amino acids of chick diets containing soybean or cottonseed meal. Poultry Sci. 31: Matterson, L. D., E. P. Singsen, L. Decker and A. Kozeff, 191. A comparison of several antibiotics as growth stimulants in practical chick starting rations. Conn. Agr. Expt. Sta. Bui. 27. Milligan, J. L., L. J. Machlin, H. R. Bird and B. W. Heywang, 191. Lysine and methionine requirements of chicks fed practical diets. Poultry Sci. 3: Richardson, L. R., and L. G. Blaylock, 19. Vita- THE work described in this paper was undertaken in an attempt to obtain further information on comparative physical, chemical and cooking qualities of eggs stored by five different methods of preservation that could be easily duplicated in the home. Methods chosen for study were waterglass, freezing, application of a commercial creamy emulsion, oiling, and coating with a plastic. The eggs were tested at the end of three and six months' storage. 1 Contribution from Montana State College, Agricultural Experiment Station Paper No. 263, Journal Series. 2 This paper represents a portion of the thesis submitted by Sister Agnes Marie Nash, F.S.P.A., to the Graduate School of Montana State College in partial fulfillment for the degree of Master of Science. Present address: Viterbo College, LaCrosse, Wisconsin. 3 Grateful acknowledgment is made to Dr. E. R. Halbrook of the Department of Poultry Industry, Montana State College, for furnishing the eggs used in these experiments and for his helpful suggestions and interest in the study. min B12 and amino acids as supplements to soybean oil meal and cottonseed meal for growth in chicks. J. Nutrition, 4: Sanford, P. E., 192a. Antibiotic and vitamin Bi 2 supplements in poultry feeds. Proc. of the Seventh Kansas Formula Feed Conference, p Kansas State College, Manhattan, Kansas. Sanford, P. E., 192b. Effect of feeding chicks combinations of vitamin B12 and various antibiotic feeding supplements. Poultry Sci. 31: Sherwood, R. M., and J. R. Couch, 19. The effect on chick growth of supplementing a vegetable protein diet with an A.P.F. concentrate. Poultry Sci. 29: 1-7. Physical, Chemical and Cooking Qualities of Eggs as Affected by Five Methods of Home Preservation 1-2 ' 3 SISTER AGNES MARIE NASH, HELEN L. MAYFIELD AND LURA MAE ODLAND Department of Home Economics, Montana State College Agricultural Experiment Station, Bozeman (Received for publication August ) EXPERIMENTAL METHODS Clean, grade AA eggs from New Hampshire hens were delivered daily from the college poultry farm to the laboratory during a three-week period. All eggs except those to be frozen were dated, numbered, weighed and marked with their individual fresh weights. The eggs treated by means of commercial creamy emulsion, oiling and plastic coating were packed, small end down, in paper egg cartons. All eggs were stored in the dark at to 6 F. and at a relative humidity of to 6%. Untreated eggs were stored in the same manner as the shell-treated eggs and were used as controls. Groups of ten eggs were used for each of the physical, palatability, and cooking quality measurements. Tests were made on the fresh eggs and on the eggs preserved and stored for three and six months. Methods used for treating eggs Waterglass. Eggs were placed in gallon Downloaded from at Penn State University (Paterno Lib) on May 9, 216