THE VALUE CF CORN FERMENTATION SOLUBLES IN POULTRY NUTRITION By SIMON TSAI-LI TSANG AN ABSTRACT Submitted to the School of Advanced Graduate Studies of Michigan State University of Agriculture and Applied Science in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY Department of Poultry Science Institute of Nutrition Approved s ABSTRACT Evidence is presented to indicate that CFS Concentrate #3 contains unidentified growth factor(s)« ^he growth-promoting effect of this factor(a) is modified by sexes and breeds of the birds, the origin of dietary fat, productive energy value of the ration and prolonged storage. The addition of CFS Goncentrate #3 to an all-vegetable-type ration significantly improved chick growth rate. Since the basal ration was nutritively adequate, this growth-promoting effect is used as the indication for the presence of unidentified growth fac— tor(s) in CFS Concentrate #3. The similar effect of CFS Concentrate #3 and dried distillers solubles in stimulating chick growth suggests that they may contain the same unidentified growth factor(s). No growth response was detected when fish meal or dried brewers yeast was used In che basal ration* The chicks were not depleted for any growth factor prior to the experiment which, in part, explains the failure of the two additives to stimulate chick growth under these conditions* Increasing productive energy from 996 to 1083 Calories per pound of ration, while the C/P ratio was kept at about 50, improved feed efficiency. With the higher caloric level, supplemental CFS concentrate #3 stimulated the growth rate of chicks. Thus, diets containing over 1000 Calories of productive energy per pound of feed are desirable if CFS Concentrate #3 or dried distillers solubles is to exert its best effect in stimulating growth of meat-type chicks. It was effective in the presence of dietary antibiotic* However, prolonged storage of the product reduced its growth-promoting value. White Leghorn, male birds benefited more from supplemental CFS Concentrate #3 than straight-run chicks of the same breed* Increasing the level of refined cottonseed oil from three to six per cent of the ration decreased growth rate of White Leghorn, male chicks* #3* The effect was overcame by the addition of CFS Concentrate When refined cottonseed oil was added to the diet of meat-type birds, they did not respond to CFS Concentrate #3* The nutrients other than the unidentified growth factor(s) supplied by CFS Concentrate #3 were also utilized by chicks. Th e v a l u e of c o r n f e r m e n t a t i o n s o l u b l e s IN POULTRY NUTRITION SIMON TSAI-LI TSANG A THESIS Submitted to the School of Advanced Graduate Studies of Michigan State University of Agriculture and Applied Science in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY Department of Poultry Science Institute of Nutrition 1959 ProQ uest Number: 10008543 All rights reserved INFO RM ATION TO ALL USERS The quality o f this reproduction is dependent upon the quality of the copy subm itted. In the unlikely event that the author did not send a com plete m anuscript and there are m issing pages, these will be noted. Also, if m aterial had to be removed, a note will indicate the deletion. uest ProQuest 10008543 Published by ProQ uest LLC (2016). Copyright of the Dissertation is held by the Author. All rights reserved, This w ork is protected against unauthorized copying under Title 17, United States Code M icroform Edition © ProQ uest LLC. ProQ uest LLC. 789 East Eisenhower Parkway P.O. Box 1346 Ann Arbor, M! 48106 - 1346 ACKNCMLlDGEIvlEl'/rS The author wishes to express the sincere thanks to Dr* Philip J* Schaible for his constant personal interest, motivation and guidance throughout this research* Sincere appreciation is due to Dr* Howard C* Zindel for the many courtesies extended, and for the constructive revi«tt of this thesis. Special thanks are due Michigan State University and Clinton Corn Processing Company for financing this research through an Assistanship and a grant-in-aid* Special thanks> are due to Dr. Erwin J. Bennie for his assistance in certain chemical analyses, Dr. Robert J* Evans and Selma L* Bandemer for their help in the determination of sulfur and to Dr* Richard U* Byerrum far his constructive review of this thesis* The author is also indebted to his fellow graduate students, Charles W. Pope, Robert H. Roberson, Hugh F. Travis, William K. Warden and J. B. Ward for their assistance and many helpful sugges­ tion** CONTENTS Page INTRODUCTION................................................... 1 LITERATURE REVIEW.*............................................ 4 A* The need of unidentified growth factor B* Conditions which modify chick response to different sources of unidentified growth factors................ * a) Body stores or carry-over effect*............... b) Sex............................................. c) Genetic background ....... ..................... d) Composition of diets*............................ e ) Interrelationships *........... f} Multiplicity* .......... g) Variability in potency*............... h) Environmental microbialpopulation. ..... i) Uncertain conditions......... 5 5 6 6 6 6 7 7 7 7 The organic and inorganic natures of unidentified growth factors* .••••............ 8 C* D* .* 4 Recent knowledge of the mineral requirements of growing ........... 8 chicks a) Potassium*. ...... 9 ...................... 9 b) Zinc ....... . 10 c) Molybdenum* d) Selenium*... ................. 11 e) Bromine*. .......................... 11 f ) Sulfur *.......... 11 EXPERIMENTAL PROCEDURE..........................................16 RESULTS AND DISCUSSIONS......................................... 26 A* B# Evidence indicating the presence of unidentified growth factor(s) in CFS Concentrate #3*..................... Experiment 1............... 26 26 Factors affecting response to CFS Concentrate #3#..... * 34 a) Sex difference ... .... 34 Experiment 2 .......... ....... .................. 34 Experiment 3«........................ 38 b) Breed difference *.... 46 Experiment 4* * .0 46 c) The origin of fat .............. 52 Experiment 5©......... Experiment 6®..........,........................ d) Prolonged storage ®............ Experiment 7 ...... e) Productive energyvalue of the basal diet®........ Experiment 8 .......................... Experiment 9 .... 52 56 58 58 67 67 72 C* Replacement value of CFS Concentrate §3 for f i s h m e a l . 81 Experiment 10.................... ............. * 81 D© Availability of vitamins contained in CFS Concentrate #3#.................. Experiment 110............... Experiment 12©............................. 87 87 91 E© Effect of supplemental sulfur on chick performance*„.... Experiment 13®............................... Experiment 14**........*................. Experiment 15. .............. 93 93 94 97 F. Influence of urea on sulfur utilization by chicks©......100 .100 Experiment 16. ............. Experiment 17........... o•.104 G* Importance of supplemental vitamin K in rations con­ taining corn and soybean oil meal as the only sources of protein*. .................. Experiment 18..©........... .107 107 H* Other experiments© . ......................... .110 Experiment 19 ............. «110 Experiment 20*......... ©HI GENERAL DISCUSSION* *.............................. .......... 112 CONCLUSION........... ©............ 115 LITERATURE CITED®............................... ©..............116 1 INTRODUCTION Corn fermentation solubles is a by-product of the corn wetmilling industry# Shelled corn is the starting material and the products made from it include various grades of corn syrups, corn starches, dextrins, crude and refined corn sugar (dextrose), refined edible oil, lactic products and livestock feeds, including corn fermentation solubles* The first operation Is steeping or soaking the corn with water which softens the kernel and removes the soluble material. Five to seven gallons of water are required for every bushel of corn# Sul­ fur dioxide is added to the water to control fermentation and pre­ vent putrefaction. compounds* This assists also in the extraction of the soluble During the process water is separated and reused again in steeping* Finally, the liquid is withdrawn and concentrated to a solids* content of approximately 50 per cent* This concentrate, crude corn steep liquor, may then eithar be sold as it is, or combined with corn gluten feed to be marketed for use in livestock feedso The technology of this process is discussed in detail by Kerr (1950), Brautecht (1953), and Radley (1954.) a A- general flowsheet of the corn wet-milling process taken from the report made by Liggett and Koffler (1948) is presented in Figure 1, Corn steep liquor has been used for the production of penicillin since 1942 (Moyer and Coghil, 1946)« More recently, it has been used also in poultry feeds, in liquid form or sprays-dried on other 2 GENERAL FLOWSHEET Shelled Corn | Stee ps | Steeped Corn Attrition . Steep Water mils ("Foos")! I | Evaporators | H# I Germ Separators! Germ r ~ -----|Heels| Degerminated Corn I Buhr Mills I |Driers| |Reels | foil Expellers"]" Starch, Gluten, Grits f Shakers" Oil. | 1Cake Ifiltersl (Mills. I fi Starch, Gluten J Bran Griis | Tablesj Starch i— Washing Filters Gluten f— *-|Presses I |Ref inery | 1Driersfcg r iTI {Kiln House) r Crude Oil Oil Cake Meal Corn Syrup, Corn Sugar Pearl, Powdered, Pure Food Starches Modified Starch House I— Thin Boiling Starches Gluten Feeds Corn Steep Liquor Fig* 1* General flowsheet of the corn wet-milling process. The flow of water is counter-current to the flow of materials as shown in the diagram. To maintain the water balance in the plant, water may be in process and storage for as long as two weeks* The water in the evaporator may come from any stage of tne process* (Liggett and Koffler, 1948) 3 corn products* When steep liquor is dried on corn gluten feed, it is known as corn fermentation solubles* When supplemental vitamins are added, the product is called CFS Concentrate #3 by Clinton Corn Processing Company* The experiments described herein were designed to evaluate CFS Concentrate #3 (CFS from hereon) in poultry nutrition* 4 LITERATURE REVI2W A. The need of unidentified growth factors. The need for certain unidentified growth factors (UGF ^reafter) by the chick is indicated by numerous^ reports* Carlson et al. (1949) found that the chick requires two factors, contained in fish meal and brewers dried yeast, respectively. Menge and co-workers (1952) reported that liver fractions, WL M and Biopar “C”, contain the same UGF as brewers dried yeast and this differs from that in dried whey or whey products* Fuller et al. (1952) found the chick required two UGF, one in dried whey and the other in fish solubles. Couch et al. (1952) also demonstrated that chicks, fed on an all-vegetable ration, need two UGF— one in fish meal, the other in dried distillers solubles and whey products. According to Camp et al. (1955 and 1957) an antibiotic fermentation product and fish solubles contain one factor, whereas corn steep liquor and dried whey supply another. Norris (1954) suggested the existence of three distinct UGF— in dried distillers solubles, dried whey product or grass juice, and fish solubles. Fisher and co-workers (1954) showed that alfalfa meal, defatted whole liver, and dried whey contribute three distinct factors toward the rapid growth of chicks. According to Scott (1955) the factor contained in dried distillers solubles is different from those in alfalfa meal, whey, liver powder, and fish meal. Tamimie (1955) stated that the organic factor of fish meal is identical to the "liver11 factor. Couch et al. (1952) reported that dried whey 5 product and dried distillers solubles contain the same factor* Three distinct UGF were recognized by the Cornell group (Norris, 1954) and four by the Illinois group (Fisher et al*, 1954$ Scott, 1955 and Tamimie, 1955)* Discrepancies as to the number of UGF may be explained in part by the faet that conditions modify the chick response to different sources of UGF* B • Conditions modify response to different sources of unidenbified growth factors* The influence of certain conditions on the chick response to UGF has been noticed by several workers* For example, Combs et al. (1954) postulated that differences in body stores, intestinal flora, genetic background, sex, composition of diets, as well as interrelationships among UGF modify chick response to such factors* (1955) made similar suggestions. of many other Petersen et al* These are borne out by the findings researchers. a ) Body stores or carry-over effect: Menge et al. (1952) found that chicks fromdepleted hens to the factor supplied by a liver fraction whereas progeny of non-depleted hens do not* respond orbrewersdriedyeast, Similarly, Johnson (1953) noted that chicks from depleted hens reacted to the dietary addition of fish solubles, whereas those from non-depleted hens did not. According to Kohler and Graham (1952) the response of the chick to factors contained in forage juice is dependent upon the of the maternal diet* make-up March et al. (1955) also stated that the 6 factor present in dehydrated green feed is carried over from dam to chick* d) Sex: Arscott and Combs (1955) reported that female chicks respond to liver products or fish solubles to a greater extent than males. On the other hand, Lillie et al* (1953) found that male chicks are more responsive to the addition of fish meal. c) Genetic background: Researches by Lillie et al* (1958) showed that differences in breeds and systems of mating affect the carry-over of the UGF of fish solubles* d) Composition of dietst Barnett and Bird (1955) suggested that the change in intestinal flora due to dietary antibiotics alters the response to UGF, High levels of vitamins, as well as antibiotics, in the diet were found to spare the UGF requirement of chick (Combs et al*, 195-4 and Arscott and Combs, 1955)* Type of carbohydrate in the ration also influences the chick response to UGF according to Hills et al. (1953) and Arscott and Combs (1955)* Rasmussen and associates (1957) found that the value of UGF in various crude sources were more fully expressed if they were used in a ration containing a productive energy of 1077 Calories per pound of feed. e) Interrelationships s 7 March et al. (1955) found out that response of chicks to either of the factors contained in dehydrated green feed or herring meal, is dependent to the presence of the other in the ration. f) Multiplicity: According to Combs et al. (1954) and Rasmussen et al. (1957) most of the crude UGF sources contain more than one factor* Condensed fish solubles contain both of the factors present in liver fraction and dehydrated alfalfa meal (Arscott and Combsl 1955)# tillers solubles contain two growth factors— "Vitamin Dried dis­ and one similar to that in liver residue (Rasmussen and co-workers, 1954)* g) Variability in the potencys Couch et al. (1954) have pointed out the existence of variations in growth factor potency among samples of different sources of UGF* Recently, Reid et al*. (1958) stated that the potency of UGF in dried distillers soluHes; decreases during storage* h) Evironmental microbial population: Barnett and Bird (1956) discussed the difficulties encountered in the study of UGF in poultry nutrition, ^hick responses varied from year to year over a three and half year period, due to changes in the environmental microbial population. i) Uncertain conditionsi Camp et al. (1957) found that the incorporation of fish solubles* dried whey or corn steep liquor in an all-vegetable ration promoted chick growth in two of three experiments. They stated "chicks some­ times fail to respond to sources of unknown growth factors*" Waibel 8 et al. (1955) also referred to "the well known, but usually unpublished, difficulties in studying unidentified chick growth factors." Since negative results are frequently not published, the complete picture cannot always be ascertained. C. The organic and inorganic natures of unidentified growth factors• It has been found that part of the growth-promoting effect of UGF is related to the mineral content of the crude sources* The nature, both organic and inorganic, of five sources of UGF has been researched by Morrison (1955), Norris (1955), and Morrison et al* (1955 and 1956a and b). Tamimie (1955) stated that fish meal contains two UGF— one organic, the other mineral. Couch et al. (1955), Dan- nenburg et al. (1955), Reid et al. (1956) and Camp et al. (1956) reported that the ash of dried distillers solubles or fish solubles stimulates chick growth to the extent of about one-half that of the intact material. On the other hand, Scott et al. (1955) did not get a growth response with the ash of dried distillers solubles whereas with the intact material they did. Briggs (1956) also failed to obtain a response by adding the ash of crude sources of UGF to a purified diet. He was critical of research workers who failed to supply levels of minerals recommended by the National Research Council (1954, NRC hereafter) in their chick rations. D . Recent knowledge on the mineral requirements of growing chick: Since the publication of the NRC nutrient requirements for poultry, further researches have been conducted with respect to the 9 levels of certain minerals which give optimum results, as well as on the possible need for other trace minerals* Some of the latter have been found partially responsible for the growth-promoting effect of the ash from certain sources of UGF. a) Potassiums Burns et al* (1953) found that the chick requires 0.23 per cent potassium in the ration* However, 0*4-0 per cent gave maximum growth. According to Leach and Norris (1958) the potassium requirement varied from 0.25 to 0.32 per cent of the diet, depending upon protein and energy levels. These levels used were higher than that recommended by the NRC. b) Zinc: Roberson and Schaible (1958a) employed a semi-synthetic diet containing ’’Brackett11 protein (isolated soy protein) and plasticcoated or glass equipment to show that chicks require zinc as a nutrient for normal growth, feather development, health and efficient utilization of feed. 0,Bell and Savage (1957) obtained a response from supplemental zinc with "Brackett" protein but not with casein in the diet* According to Norris et al. (1958) zinc naturally pre­ sent in "Brackett" protein is not utilized by the chicks whereas that in casein is* Roberson and Schaible (1958) as well as Norris and associates (1958) noted that chicks require zinc supplementation only if purified ingredients are used and certain protective measures are taken to prevent contamination. According to the former, the aine requirement of growing chicks is 20 ppm which was confirmed by 10 the latter workers* 0*Dell and Savage (1957) found that zinc was able to replace part of the mineral supplied by dried distillers solubles and Norris et al. (1958) stated that the zinc and potassium contents in the ash of UGF supplements do. not fully explain their growth-promoting pro­ perties* Beid et al* (1958) also stated that the growth-promoting effect of dried corn distillers solubles cannot be entirely accounted for by its ash content. c) Molybdenum: Keid et al. (1956) and Kurnick et al. (1957) employed a diet containing "Brackett" protein and reported that chicks require molyb­ denum as a nutrient. The growth-promoting effect of supplemental molybdenum at 0.0216 ppm was found equivalent to that of the ash of dried distillers solubles (Reid et al., 1956) although intact product promoted a faster growth rate. When molybdenum was supplied at 0.03 ppm, growth rate almost equaled that of 3 per cent intact dried dis­ tillers solubles (Kurnich et al., 1957). Higgins et al. (1956), and Leach and Norris (1957) produced molybdenum deficiency among chicks fed casein diets only after the addition of tungsten as an antagonist. According to Norris et al. (1958) the failure to obtain a response to supplemental molybdenum was due to the availability of this element in casein and most of the molybdenum of "Drackett" protein is not utilized by chicks. The chick1s requirement for molybdenum without the antigonist— tungsten— was suggested as not more than 0.24- pg/kg of diet* u d) Seleniums Schwarz et al* (1957) identified selenium as an integral part of "Factor 3" which was found highly effective against exudative diathesis in chicks. To produce this syndrome a torula yeast diet was employed (Schwarz et al., 1957$ Scott at al., 1957 and Patterson et al. 1957)* Nesheim and Scott (1957) reported that 0*1 ppm of selenium prevented the development of exudative diathesis and stimu­ lated growth in chicks* However, when casein was used as the main source of protein in the diet, the addition of selenium did not in­ crease the rate of chick growth (Norris et al., 1958)* Since natural feedstuffs generally contain ample selenium for growth (Nesheim and Scott, 1957) and this element becomes toxic at ten ppm level (Carl­ son and LeitiS:, 1957) it is doubtful that any benefit will result by adding selenium to practical rations* e) Bromine: According to Bosshardt et al. (1956) the incorporation of eight or 15 ppm of bromine in a semi-purified diet promoted chick growth in three out of four experiments. Norris et al. (1957) reported that ten to 20 ppm of bromine improved chick growth slightly in two experiments^ in two others, a slightly depressed growth was observed. Since bromine is plentiful in natural feed ingredients, especially salt (Ucko, 1936), whether or not chicks require supplemental bromine is still uncertain* f) Sulfur: The ability of ruminents to utilize inorganic sulfur has long been known* More recently, Hale and Garrigus (1953) reported that 12 yearling wethers use elemental sulfur for cystine synthesis in the wool# To prove this, radioactive sulfur was administered as elemental sulfur or sodium sulfate# Albert et al# (1955) and Stark et al# (1953) found the inclusion of 0#2 per cent inorganic sulfur in diets sup** plemented with urea, promoted lamb growth* Employing an artificial rumen, Hunt et al. (1954) noted that elemental sulfur or sodium sulfate increased the micro-synthesis of riboflavin and vitamin and the utilization of urea# The latter was more effective. The increased synthesis of vitamins due to sulfur was attributed to the need of the microorganisms in the rumen for this mineral# The first report on the effect of sulfur in poultry feed was made by Philips and co-workers (1921)*, Addition of elemental sulfur to the feed and copper sulfate to the drinking water caused a 19-per~ cent growth increase over the basal at the age of eight weeks# The level of sulfur was calculated to be 1#6 per cent of the ration* The basal ration contained no added mineral except 2*44 per cent ash from an unknown source# The growth rate of all the birds was consi­ derably lower than present* in a basal ration* Therefore, a knowledge of the nutrient contents of the basal rations is exceed** ingly important* a) Nature and purpose of the basal rationss A review of literature shows that the growth-promoting effect of UGF Is not necessarily dependent on the use of synthetic diets5 under certain conditions it can be detected with diets of natural ingredients* Since CFS Is manufactured for commercial use and certain vitamins are added to assure guaranteed levels (Table 2), it was decided to use a basal ration of natural feed ingredients* To assure the adequacy of the basal rations, two criteria were set up* First, the rations must contain all known nutrients at 19 a. § ft xO (0 t w § •H •8 p vr^vr\VP\u-^o O O O O O O u^u^u~\0 O Q O O -© O O H i —I i — Ii I H i —I i —| i —|i — I rH i —I i — 1i —I i — li—I i —li— IrH i —IH i —Ii — IrH S THE CHICKS, DURATION AND REPLICATIONS OF EACH EXPERIMENT •d" 1 I rH CM i— ICr'\ rH i— 1 0r''\ -Ch r— I t— I r— I i— I rH i— I CM CM C*^\ -d" *© 2? © ©*53© ba444444444444 © © © © CD 0 CD 4(4D 404 4I4D 4O4 44P34444444544P3 ID O (D © © © © © > © VO I I} >»t5 © © © I © © © T3 CM ££ lilt i— Ift ft P h C O rH i— I ft g CO © M sg 4O4 -H 6 40 ft> tZ> • • 40 CO CO O • • k' S© b © © d XJ XJ i—1CM CM Sft is 1© © tr H ■d -fp © a CO a *8 • • * 03•1-5 O. • • • 15 2s CO © ft 00 43 •b HO D *h sH -. tZ* >to• 4f0t 40 40 CO CO a o a a 'd £ i —li —l -p 44 o L 44 b ® cd © © © ^ © © © © © © © © I 2 *2 JS J2 |2 I I Ir£H r5l H rH iH H 2 £ o 2 o o h fn ^ C5 c5 c5 © CO -©■ CO CO S £O -p -p 43 43 ^ p • to• D P P io + +•* CQ Eft COCO CO 01 a a aa a s CO *bD H ‘hf) H ft ft h5 o 5 >» b b ©b ©b ©b ©b ©b ©b ©b ©b ©b©«©b©b©b©b © b © © © xs d'd'd'd’d ’d x S ’d'd'd'd'd t ts d xs"d rH ( — IH i —Ii — Ii —I i —I i —Ii —Ii — Ii — | O i —Ii —I '-Oi —Ii —I i —I■ —1 ft s ft ft © © © © © © i l 1 I—1 © —ii—ii © — © I ftI © © © i—1i —I i ft ft ft I—II—Ii i —1 ft ft r©H i—! © cd © - CO © ( © © © p P © P © -p -p *SJH*d fcrHa a CO CO to a a a a rH a a a 0304030303030305 •03 •03C3P3P3P3P303 03• CL, • X,pL,P-,fft0^fft (4, ft ft ft ft ft ft ft is * * * * 1 LT\\0 in-co o o pj * * * * * * i-2r X CM C^-3" xT r' \vo {>- CO 0 \ 0 ( — Ii —I H i —I i —I i —Ii — ti—I CM Note Birds of experiments with asterisk poultry house 500. P w4 ft 03 -QN -^•Jd'COCMvOrHCOCM c^3- cox op X- cp-P r —Ii —Ii —IiIi —Ii —Ii —Ii —i W)g.O QN t> Cr'i £N- ON H 3 H CM cvi r\ \Q V O V O V O \O V O \O V O HI -Sp p 3 O e-l g •r op £ 0 P< O A X E~t g 0 0 iM 'ge rH O -0 P £ 0 0 c d Xco XJ £> £ 0 £< H ■8 •H -H SJ E o Hi rH Q o g OQ M s e» e o HI tu E-i 0 pH B o 0 0 o £ g c/5 •*-P ^ ft p (H r0l -WH T!N=fc.VV<&. 0 t3 0 OP C^ <*5 £ S -o 0 + + + X0J «0H 3 S *H £ 0 O H Q P £Q £O £ OS O £ O ^ \ P. S n n c*5cn n . 5j 0 x*1 0 £ 0 £P -0 0 • £ 0 -P § £ £ 0 bjD X,ON X -X 0 0 £ rH 0 0 £ •P 0 •sH 3 * Xo o £ cO Pi 0 Pi 0 0 0 X£ 0 •H 3: X 0 Q X i —1 0 O -H o rH 0 P. 0 0 £ X0 X o o .H 0 1—1 0 PU 0 £ *£H £ £ ■g O H, X * 30 of the requirement (0*24 ppm)« It is apparent, therefore, that the basal ration does not need supplemental cobalt or molybdenum. Con­ sequently, the growth promoting effect of CFS or dried distillers solubles is due to factor(s) other than these trace minerals. The lack of response from adding dried distillers solubles to the ration containing CFS is evidence that three per cent of either ingredient supplied enough of the factor(s) required by chicks for rapid growth on this basal ration. The failure of fish meal or brewers dried yeast to increase growth rate does not mean that these additives contain no UGF. A high level of soybean oil meal was used in the basal ration and this may have prevented fish meal from showing any growth-promoting effect. Hill and Briggs (1950) reported that soybean oil meal con­ tains UGF, and Savage et al. (1950) showed that soybean oil meal and liver residue contain the same factor(s). At the same time, Petersen et al. (1955) found that fish meal and liver residues contain the same growth factor(s). More recently, Reid et al. (1958) found that the extraction of soybean protein with isopropanol increased the response of chicks to fish solubles but showed little influence on the growth-promoting effect of corn dried distillers solubles«. Thus, unwashed soybean protein contains the nfish factor" but not factor(s) supplied by dried distillers solubles,. The basal ration used in this experiment contained more than 36 per cent dehulled soybean oil meal and, thus, may have had enough "fish factor". Under these conditions, any fur- 31 CN-=* -3" Statistical results for body weight, Experiment I: rH On CM 8 SCQ O -3- CO S -=J- 8 O -=r rH CM rH 0- OO OH O OH CO CO 0 o c o 52; 44 (D 1—1 CD > 0 1—1 NtH 't R rH pH Cti 32 ther addition of a carrier of such factor would not be expected to increase chick body weight. Menge et al. (1952) reported that birds from non-depleted hens showed no response to brewers dried yeast. Since commercial chicks were used in this experiment, the hens were not depleted for such growth factor(s). No response, therefore, would be expected from the supplementary brewers dried yeast. assumption* The results substantiate such 33 TABLE 7 THE TRACE ELEMENTS SUPPLIED 3X BASAL RATION OF EXPERIMENT 1. IN comparison w i th chick requirements* Trace mineral ^ or unit of the ration Experiment 1 Requirement K, £ 0*25 - 0.32 0.91 Co, mg/lb — Cu, mg/lb 0.9 Fe, mg/lb 9 Mg, mg/lb 220 806.73 Mn, mg/lb 25 30.29 M o , ppm 0.2^ S e , ppm 0.1 - 0.2 Zn, ppm 28 - 33 * See Table for sources of Information See text 2.97 21 See text 0.33 36.6 34 B,. Factors affecting chick response to CFS Concentrate #3• a) Sex difference : EXPiiRIMiSWT 2 This experiment was designed to study the effect of CFS on the growth rate of straight-run, White Leghorn chicks. The composition of the basal ration is presented in Table 8. Three levels of CFS were added by replacing corn and soybean oil meal so that the protein level was equalized to that of the basal. The results are presented in Table 9* Since the feed consump­ tion was recorded by groups, and only one group of 15 birds was used for each treatment, the feed efficiency cannot be treated statistically. However, individual weights were taken and analyzed for variance due to treatment. The variance was found not significant. Therefore, under the experimental conditions the incorporation of CFS at levels used did not affect chick growth. 35 TABLE 8 THE COMPOSITION OF BASAL RATION FOR EXPERIMENTS 2 AND 3 Experiment N o . ________ 2 Ration ] Ration A 2 Ingredients Lbs./Cwt Ground yellow corn Dehulled soybean oil meal Refined cottonseed oil Methionine Ground limestone Dicalcium phosphate Iodized salt Trace mineral mix Vitamin and antibiotic mix 55.to 36.73 3.00 0.05 1.60 2.00 0.50 0.10 0.62 55.61 36.70 3.00 0.05 1.60 2.00 0.50 0.10 0.44 51.99 37-32 6.00 0.05 1.60 2.00 0.50 0.10 0.44 Calculated values; Protein, $ Productive energy, Cal/lb Metabolizable energy, Cal/lb 23 956 1240 Trace mineral mix MnSO^ (70fo) FeSO4 *7H20 CuSO^KoO CoSO^TH-O ZnCl^ DehuXled soybean oil meal (carrier) 23 958 1243 gm/100 lb ration 10.9769 4.5159 0.3565 0.0431 0.0059 29*4607 23 1003 1320 36 TABLE 8 (cont*d.) THE COMPOSITION OF BASAL RATION FOR EXPERIMENTS 2 and 3 Experiment No Vitamin and antibiotic mix 2 3 gm/100 lb ration Vitamin A (10,000 IU/gm) Vitamin D (3*000 ICU/gm) Alpha-tocopherol acetate (20 gm/lb) Menadione (8 gm/lb) Riboflavin Calcium pantothenate (32 gm/lb) Niacin Fyridoxine hydrochloride Choline chloride (25%) Folic acid Biotin Inositol Vitamin B^5 (6 gm/lb) Terramycin (10 gm/lb) 40.00 9.00 45.36 __ 0.10 28.35 1.00 0.23 90.00 0.08 0.01 0.01 45.00 22.50 £jo*o o 9.00 4.60 5.68 0.10 28.00 1.00 0.23 45.00 0.08 0.01 0.01 45.00 22.00 37 TABLE 9 THE EFFECT OF CFS ON THE PERFORMANCE OF STRAIGHT-RUN WHITE LEGHORN CHICKS* (Experiment 2) Treatment Av* final weight, gm Feed/gain Basal 195 2.05 1.5# CFS 199 1.99 3.0# CFS 196 1.97 6.0# CFS 19*f 1.98 * Two-day_old, straight-run, "White Leghorn chicks -were subjected to one week standardization period before the two week experiment. One group of 15 birds with an initial average weight of 67 grams per bird was used per treatment. 38 EXPERIMENT 3 This experiment was designed for two main purposess First, to study the effect of fat levels on chick response to CFS. Second, to find out if a standardization period is required for the detec­ tion of such a response* Two-day-old, male, White Leghorn chicks were wingbanded and divided into three groups* Group I s Birds were standardized on basal ration A for six days. The composition of this basal Is presented in Table 8. On the seventh day, the chicks were distributed into three lots as des** eribed. Two levels of CFS were added at the expense of corn and soybean oil meal, so that each ration contained 23 per cent protein and three per cent added fat. Group Zt Two-day-old birds were assigned randomly to their respective treatments without a standardization period. The rations used for Group 2 were the same as for Group 1 with one higher level of CFS added. Group^s as in Group 2o Table 8. Birds were assigned to their respective treatments The composition of basal ration B: is presented in Three levels of CFS were added at the expense of corn and soybean oil meal so that each ration contained 23 per cent protein and six per cent added fat. The results are shown in Table 10. The variances of final body weight due to treatments for Groups 1 and 3 were significant at five and one per cent levels, respectively. This indicates clearly that 39 c •H cfl lKE EFFECT OF CFS CN CHICK GROWTH AND FEED EFFICIENCY AS INFLUENCED BY STANDARDIZATION PERIOD AND FAT CONTENT OF THE RATIONS (Experiment 3) 73 0 EH CD Pt< • -3- o co 00 £V C"- rH CO rH CV. 07 07 07 C7 00 *8 00 p- G 0 f t 0 G O »H 73 cd O CM 73 O •H G 0 P CO N •H 73 a Xi M-l l * r 0 £ 0 cd 73 G 0 • r H P P •rl « rH -Aj 00 rH g G P 1 —I cd tsj 73 . G hJl cd O G & 73 rl p •rH cU 73 § 0 P 0 ji o Xi if 0 0 Cd X5 P •H JS P cd rH MO 0 0 ft C7 ■8 G •H cd •s 5 G o o p G P G GO O O 0 G O OH cd AjMO -3MO A± MO O O 0 G O • r H P •H P G G 15 «H «h 0 73 G •H CQ I ■—1 a, E-> o 0 G 0 73 73 cd £ cd 1 73 p G ■s .s ft •bd H cd «m G G 0 O = t JS P 0 P 73 07 07 P MO GH •cd bfl 07 ch cd g cd 73 0 CO 73 G •H cq 0 g £ O K 0 u7 * 1 —1 O • 07 0 • MO 40 g -H (0 CP OP • bo *©8 C\2 P*H ■cj• rH O CO• vp C>•- rH rH 8 • r l G © P- CO g & 73 •H 4-5 P < § ,G • 3 © N •H VT\ 0^ -8 rH rH W P G © CP 1 P< g g o V®. ’S' « o © G en ■s o G C5 © cO CQ vp Experiment 3 8 O iS El "S’ i—I rH >P {S* -p X cO .5 C*3 VTj c\j rH C 0 0 £ 0 XI &-» 0 0 a 0 o SH rH O J-i -p 0 O O 0 0 £ -P 0 0 U £ 0 0 f—1 H 0 5« 0 iH VOl & o y-« a ■p © £ 0 © J-t 1H 0 ! > 0 i—I •P 0 •4 > 0 rH 1$ 43 of vitamin K and choline, but not of vitamin E. Xn Experiment 2 menadione was not added to the basal ration and addition of CFS did not promote chick growth. On the other hand, the basal ration of Experiment 3 was supplemented with menadione and CFS stimulated chick growth (Group l) * No supplemental vitamin K was added to the basal ration of Experiment 18, As a result birds suffered from hemorrhagic conditions whether CFS was incorporated or not. Consequently, vitamin K content of CFS cannot explain the difference observed in Experiments 2 and 3* The choline requirement for chicks recommended by NRG is 720 mg per pound of ration. Yellow corn contains 200 mg and dehulled soy« bean oil meal 1500 mg choline per pound, so the basal rations in Experiments 2 and 3 supplied 857 and 759 mg- choline per pound of ration, respectively. These levels are ample to meet the chick requirement for this vitamin. The results obtained in Experiment 6 also indicate that there is no need to add more choline to a ration containing 750 mg per pound*, Obviously, the choline content of CFS is not the answer for the discrepancy observed, Sex-differences in response to UOF are well illustrated in the review of literature. Since unsexed TTaite Leghorn chicks were used in Experiment 2 and only male chicks of the same breed used in Ex« periment 3* the influence of females; in the former case may account for the difference observed in chick response to GFS, The growth depression observed with the addition of refined cottonseed oil in Experiment 3 (Group 3) cannot be interpretated as 44 due to a deficiency of folic acid in the basal ration* March and Biely (1954-) found that the addition of cottonseed or herring oils to a ration depressed chick growth while tallow increased growtho The growth«depressing action of oil was counteracted by the addition of folic acid* It was found later that such detrimental action was due to the overheating of herring oil (March and Biely, 1955)• When fresh oil was added to a ration suboptimal in folic acid, no depression in growth was detected* The folic acid contents of the above basal rations was not reported* However, symptoms of folic acid deficiency were observed among birds fed the basal ration* Based upon the author*s calculation the basal ration was marginal in folic acid according to HRG recommendations* Ration B of Experiment 3 is calculated to contain 1*54- mg folic acid per pound of feed, of which 0*8 mg was supplied by the vitamin mix* This level is far above the chick requirement (Table 6)* The addition of six per cent CFS supplied only 0*036 mg folic acid per pound of ration which in comparison with that already provided was almost nil* Consequently, while it cannot be assured that the cot*» tonseed oil used in this experiment was overheated or not, it is certain that the depressed growth rate was due to the added cotton** seed oil per se. Since the basal ration was not deficient in folic acid, the growth stimulation which occurred with six per cent of CFS was not due to its folic acid content* It appears, therefore, that CFS aided male White Leghorn chicks in utilizing the increased level of cottonseed oil* Further, the data indicate that, under Ur5 these particular experimental conditions, male White Leghorn chicks cannot tolerate six per cent of refined cottonseed oil in the diet without reducing the growth rate® 46 b) Breed differences EXPERIMENT 4 Experiment 3 showed that an increase in dietary fat improves chick response to the addition of CFSo The present experiment was designed to compare CFS with a known source of UGF, menhaden fish meal, in a ration containing six per cent of added cottonseed oil* The birds were standardized on basal ration for eight days, after which they were allocated, as described earlier* Three repli­ cates of ten birds were used per treatment* Distilled water was provided* ration is given in Table 11. The composition of the basal Two levels of fish meal or CFS was added singly or in combinations* The calculated protein and fat levels of all diets were equalized to that of the basal* When me­ thionine was used, 0*05 pound was added to each 100 pounds of ration* Individual initial, third and eighth week week weights were taken* The average initial weight was approximately 82 grams per bird for each pen* Group feed consumption was recorded* The results are presented in Table 12* The analyses of variances showed that the final body weights and feed efficiencies up to eight weeks due to treatments were statistically significant at five and one per cent levels, respectively. No differences of consequence were detected at the third week* The addition of fish meal at the 5*6 per cent level depressed chick growth (P^/‘0*05) and reduced the feed efficiency (P^0*0l) ^7 TABLE I I THE COMPOSITION OF BASAL RATION FOR EXPERIMENT h Ingredients Lbs/Cwt Ground yellow corn Dehulled soybean oil meal Refined cottonseed oil Ground limestone Dicalcium phosphate Iodized salt Trace mineral (Delamix)* Vitamin mix 52.-40 37.60 6*00 1.32 1.88 0.50 0*10 0*20 Calculated values: Protein, $ Productive energy, Cal/lb Metabolizable energy, Cal/lb 23.25 1010 1328 Vitamin mix: gm/100 lb ration Vitamin A (10,000 IU/gm) Vitamin Do (1*500 ICU/gm) Alpha-tocopherol acetate (20,000 ICU/lb) Menadione Thiamin hydroxychloride Riboflavin Calcium pantothenate Niacin Pyridoxine hydrochloride Choline chloride Folic acid Inositol Vitamin (1 mg/gm) * See Table 13 33.37 23.00 4.60 0.20 1.00 0.30 0.60 1.50 0.20 25.00 0.25 0.10 0.60 kQ u vO < —I A p G PQ4> 0 £ ,G p o o O H V O Q W n J - CO O n ( ^ 4 O O f-IO x O O O O N r H O N rH o u *o o • *•••••«•• NWNHWNCMHCMH 00 G •H cd O i —I VA A - © 0) • • O *A THE EFFECT OF CFS, FISH MEAL AND SUPPLEMENTAL METHIONINE ON CHICK PERFORMANCE (Experiment k) CM (A 0 0 £ T5d U r j 0 I O N N H 4 - 0 O n cm CO V O 0 s- A - A - A - A - A - A - vo i i rH rH v.„o-> 'j i—I i—I i—I i—| i—I (—| CA CD • cvi e 0 hn 0 J3 • » p p: wi £ •rH 0 £ 0 % U 0 4 ^©4 © £ ru3 o CA VO CO t N ( O N N O (O H H O H v O C p O l N r l ' A t N vA l -3F A -3“ a i—I t —I i—I r l i—I i—I H CD -3" 4 IN- i—I i—I i—I p CO CO O A - CM A - V O r H A - ^ O S rj N C K H W N H O J Q C A ^ f -3" ( A -3" -P ca P bfl •H 0 £ TH3 O P 0 0 s 5 •3O in rH B A- Sr 1A A- g O 't i A- Pi -1 1 -=f CO § A- £ rH vO CA JH SJ £ ■3 0 g © s a? 0 &H E P0i S + +o + •rH P«4 . g S v 2 + ho o E S »— i :50ifC CVICD VO 1 0^ ^© \c 2b -\ VO VO W VO cd • • • ro c m * A ca • • a - c m *a # cd • CQ va 0 -P i —I G 0 0 U VC. vO • A cd 3> H CO 5? rH O -H P 0 •rH P 0 P CO I P 0 0 0 P G f-i 0 £H S VA 49 by the end of this experiment. This detrimental effect was not detected when a lower level of fish meal was used* The addition of 0.05 per cent methionine to the diet did not improve feed efficiency* CFS at both levels, fish meal at the lower level, singly or in combinationsj and supplementation of the basal ration or ration containing 7.2 per cent CFS with methionine did not alter chick performance. Experiment 3 showed that addition of oil improved the response of the birds to CFS. CFS was found to aid chicks to utilize this added oil. However, six per cent of cottonseed oil was also added to the rations of Experiment 4* chick performance. let, the addition of CFS did not improve Judged by the satisfactory growth rate of the White Plymouth Rock, male chicks, it can be said that their tolerance to cottonseed oil, or ability to utilize this refined oil, is much higher than that of White Leghorn, male chicks in Experiment 3* As a result the difference in response to CFS of these two experiments indicates a breed difference in the tolerance toward refined cot­ tonseed oil* Data from this experiment do not permit the author to distin­ guish whether the depressed chick growth was due to an imbalanced nutritive condition created by adding 5*6 per cent fish meal, or that fish meal contains a toxic factor which exerts its effect at certain levels. For instance, methionine is needed by chicks for growth, but at higher levels it depresses growth (McKittrick, 1947)* Alfalfa meal contains UGF which promotes chick growth at certain low 50 VO vO 'A rH CM S' ♦ •A* si • * vr\O- S' C\2 CO O• CM 'A O CM ■3O CM iH O• CM iH CM O U o o g o CM -P Statistical results, Experiment P ON * ON CA G O f-. 0 O *rl -P G OO «H 0 *08 ) •C8 D £ si £ ON ON CO -P G CD E -p c d CD G £-1 £ O ON G c d CD (—I 0 t 0> VA 0 t 51 levels (Fisher et al*, 1954-) but at higher levels, its inhibition factor (saponin) depresses growth (Peterson, 1950 a and b). 52 <0 The origin of fat: EXP3MMENT 5 This experiment was designed for two main purposes; First, to study the effect of CFS when the calculated levels of protein, ribo­ flavin, niacin, pantothenic acid, choline, methionine and added fat were equalized* Second, to investigate the effect of supplemental sulfur on chick performance* 4s mentioned earlier sulfur dioxide is added to control fermentation and prevent putrefaction while the corn is being soaked with water in the wet-milling process* Consequently, a certain amount of sulfur is present in corn steep liquor* Three levels of CFS were incorporated in the basal ration (Table 13) in such a way that all the rations contained the same calculated levels of protein, riboflavin, niacin, pantothenic acid, choline, methionine and added fat. Five graded levels of sodium sulfate (ifa2s o p were added to the basal ration* The results, presented in Table 14 show no appreciable changes in body weight and feed efficiency due to treatments* 53 TABLE 13 THE COMPOSITION OF BASAL RATION FOR EXPERIMENT 5 Ingredients Lbs/Cwt Ground yellow c o m Dehulled soybean oil meal Refined cottonseed oil Methionine G round lime stone Dicalcium phosphate Iodized salt Trace mineral (Delamix) Vitamin and antibiotic mix 57.00 35.31 3,.00 0.08 1.30 2.30 0.50 0.10 0.41 Calculated values: Protein, $ Productive energy, Cal/lb Metabolizable energy, Cal/lb 23*02 965 1248 Trace mineral (Delamix)* Mineral________ $______ Present as: Manganese 6,000 Iodine 0.120 2.0 Iron 0.200 Copper 0.020 Cobalt 0.006 Zinc 27.000 Calcium Potassium** Hang anou s oxide Potassium iodide Ferrous carbonate Copper hydroxide Cobalt carbonate Zinc carbonate Calcite stearate continued At 0.1$ level it supplies 27 mg/lb 0.54 mg/lb 9 mg /lb 0.9 mg/lb 0.09 mg/lb 0.06 mg/lb 0.027 i> 0.0000336 of of of of of of ration ration ration ration ration ration 5^ TABLE 13 continued Vitamin and antibiotic mix Vitamin A (10,000 IU/gm) Vitamin D (3*000 ICU/gm) Alpha-tocopherol acetate (20,000 ICU/lb) Menadione (8 gm/lb) Ribo flavin Calcium pantothenate (32 gm/lb) Niacin Choline chloride (25$) Vitamin (6 mg/lb) Terraxpycin (10 gm/lb) * Briggs et al., 195& gm/lOO lb ration 40.00 13*00 4.60 2.84 0.20 5*00 1*50 50.00 47.00 22.00 55 TABLE 14 THE EFFECT OF CFS AX'© SODIUM SULFATE OH CHICK PERFORMANCE* (Experiment 5) Final Av. wt. gm Treatment Feed/gain Part 1 - Effect of CFS Basal 1.5# CFS 3.crfo cf s 6.0# CFS 201 212 208 206 1.81 1.76 1.90 1.85 Part 2 - Effect of graded levels of sodium sulfate (NaoSOn) 0.004$ 0.008# 0.016$ 0.250$ 0.500$ * Na^SO^ Na2S0^ Na^Oj. Ha2S0£ Na^O^ eq eq eq eq eq 0 .0009# 0.0018$ 0.0036$ 0.056 $ 0.113 $ $ S S S S 200 200 220 192 220 1.91 1.88 1.75 1.90 1.78 Qne-day_old, straight-run, W.P.R. chicks were subjected to 1-week standardization before the 2-we ek experiment. One group of 10 birds, with an average initial weight of 70 grams per bird, were used per treatment 56 EXPERIMENT 6 This the basal experiment was divided into two parts.In ration and the manner in which three levels of incorporated, were the same as in Experiment 5* the firstpart, CFS were In the second part, graded levels of choline were added to the basal ration in order to find out if a higher level of this vitamin was required* The results are presented in Table 15* The analysis of variance indicates no significant difference in chick weights due to treatments* Therefore, part 1 of Experiment 5 and 6 agree to each other* The failure to step up growth by per pound adding from88 to 117 mgcholine of feed proves that the 752mg supplied by the basal ration is ample to meet the chick1s requirement for this vitamin* It further confirms the conclusion in Experiment 3 that the observed growthpromoting effect of CFS was not due to its choline content, since the basal ration of that experiment contained 759 mg choline per pound of feed* Experiments 4> 5 and 6 show that the addition of CFS to a ration with adued cottonseed oil did not affect chickperformance*However, when crude corn oil or animal grease was used, as in Experiments 1, 7 and 9> CFS promoted chick growth (Table 23 )o Hill et al* (1953)> and Ascott and Comb (1955) found that the type of carbohydrate in the ration influences response to UGF* Possibly the origin of fat does also* 57 TABLE 1 5 THE EFFECT OF CFS AND GRADED LEVELS OF CHOLINE ON CHICK GROWTH* (Experiment 6) Treatment Average final weight gm Part 1 - Effect of CFS Basal 1.5# CFS 3.0# CFS 6.0# CFS 218 218 215 215 Part 2 - Effect of graded levels of choline (Basal contained 752 mg/lb) 88 mg/lb 179 mg/lb 269 mg/lt> 723 mg/lb 1177 mg/lb 216 207 216 206 21^ * One—day-old, straight-run, W.P.R. chicks were subjected to a 1-week standardization before the 2-week experiment. One group of 10 birds, with an average initial weight of 69 grams per bird, were used per treatment. 58 d) Prolonged storage: MFmmsm 7 This experiment was designed to study the effect of CFS and fish meal on chick performance when added to rations containing different levels of energy— the C/P ratio (productive energy per pound of feed/per cent of protein) being kept constant* The compo** sition of the three basal rations is presented in Table 16*, Basal ration A s Corn oil was added to this ration to give 23 per cent protein and 1009 Calories of productive energy per pound of feed. After the birds reached the age of five weeks, the pro­ tein was adjusted to 20 per cent and productive energy to 1043 Calories* Basal ration B s The composition of this ration is the same as ration A, except that Soka-Floc replaced Terramycin* Basal ration Cs this ration. Corn oil and Terramycin were not added to It contained 21.6 per cent protein and 935 Calories of productive energy per pound. The protein was reduced to 18*6 per cent and Calories increased to 972 after the birds reached the age of five weekso Although there were differences in protein and energy levels among these three basal rations, the C/P ratio was kept at appro** ximately 43 d 52 for the periods from 0 to 5 and 5 to 8 weeks, respectively* CFS and menhaden fish meal were added to basal ration A and B at the rate of three per cent singly or in combination* Three per 59 m 44 is rH •rl O O - CM A - 00 CO • • O -=1- 1 G U O O I I A- CM WO 1 I I CM O O O O vO vO O A rH i-i -3VO O _ * CM rH •i —I CM CM H CMA H I H CM O O O OD N O -P G o • JG P ■H & £ £ A O £ CM A - Q I o vO & A W ,0 PI rH G I I I O O VO VO • • • • • rH CM O O O CM -=t • CO ACO r-i OV-3- rH CO CM dp o o o O VO O vo O v S H H Io A C A r~t A - • ^3- rH CM O O O O O CM CA CM i—I VT\ i —l CO M dF o o o o vCo O vO O A rH rH • • * • I * # • * * * CM S O v O O 1 O rH CM o O O a (rv -3- O O A CO NO O Ov A CM & CA O CA CA CM i — I -it- i —I U O O X (H PI3 S CO t A CM A- O (OCO CM o O O O vo VO O A rH rH O 'S I o 1 rH CM O O O O O A ^? O vo a) ivo o o A I O -ocr >h• -cr • • •|♦| • • • * » O ON A vo CM CM (A 8 iO —I A i —I CM ov ON • A o A CM A O -sr A O O O VO O A rH H • • • • • I • *» • • CM VD vo O O a I rH CMO O O CMrH a 4 0 E I—I c o (0 § •rH ■& $3 "g 0 -H H 0 XZ O G G Pi O cd CD rH 13 rp *H 0^0 O i —I W P rH 0 P cd O CO i —I 0 in 0 4 4 > Q, s P O to XI P -3 0 G, rH * §3 h c •rH e •H TO E 4 TO O 0 I J 0 d N "S3 0 r H o U 4 4 G G X? G rH o cd p ^ ? G CD G 0 0 O !H 4 3 u ■H C 5 Q O £-*CO (5 Q H &■* > G O H O •HEW i—1 bO * U x> 0 i—I G bO 0 G p" 0 cd • G o TO 0 O S-. 0 X> rH hO f* % m 0 o c(Sd3 JQ C •rl •H rH 0 P i—1' H cd O rH H •rl 0 • G P CO § P TO CO O o O G IX| o fH Pl, P o 0 * Table 13 ** Table 5 \o A O v O O ^ H H I I I I 4 c^co CO •H O O A CM VO CA 1 m CM A 60 cent CFS was also added to the basal ration C* The protein level of each experimental ration was equalized to its corresponding basal ration* The results are presented in Table 17 and statistical results f ollowed * The addition of corn oil (rations 9 vs 5) increased the final weight of the chicks (P<^ 0 o05) and improved their feed efficiency ( P< 0 * 0 1 ) * The latter was accomplished by a reduction in the amount of feed consumed per bird as well as improved protein efficiency (P < 0 , 0 l ) * The addition of CFS to the ration containing corn oil (rations 6 vs 5) improved (P week* 0o05) chick growth rate only to the fifth The particular lot of CFS used in this experiment had been stored at room temperature for more than a year with the hope that more uniformed results could be obtained by using samples from the same batch* It is quite possible that the potency of CFS decreased after the prolonged storage* The addition of CFS to rations without corn oil (rations 9 vs 10) did not influence chick growth at the fifth and eighth weeko However, it improved feed efficiency at both intervals (P<0*0l), and protein efficiency at the fifth week (P*£l0*0l)* promote growth when added to this ration* CFS failed to The nature of the basal rations, thus, influences the response of chicks to CFS* The addition of fish meal to the ration with corn oil (rations 7 vs 5) improved chick growth only at the fifth week (P*<,0*05)* 61 ’S'S'S eh f©t 0 g p a h) 34 o O ft vo O rH CO C O CNM-M Onv o MCO MI CM CO MVCQ M Onft"I NO -r OH s COON C VCO MVCO M( CNM-0 C0 M C rO H r' Hn-v O I N CM CM- P I § cO 0 u THE EFFECT OF CFS CO CHICK PERFORMANCE AS INFLUENCED BY DIETARY FAT AND ANTIBIOTIC (Experiment 7)________________________________ p co < D w 0 ® N-j- Q CO VO o o o o CO CH Mr O ON O•r * H•O CM CM CM CM V 40 o HO♦ ^ i—Ii —0 iv CM • * • CM CM CM CM r HO N vo • CO • CM CM S•f S • S•f• ft £ V O r H r H VO VO VOVQ £N4-4 • •4•4« o o o o u 0 ft • * o o *0 8 ft ft CM 00 * ft 0 u 0 ’Xi •rl ft -4 0 00 o0 ~VOO V O ON N C M C M C M C M ( —Ii —Ii—Ii —I O NVCO -IV O C Mi —4O i —I CO OCO C C O P0 0 3 E u b) © ft CO ft ft COQO O Oi —I UV ON0 ON ONC OO n vo VO ft- CO COMO rH VO On Ov O O COVQO 0 0 On ON co-S fiQr O co o o o o C OO n 00 rH rt H OO n H CO f C ft"ft • • • • o o oo • CM • • H *OH t In•-o(N-o* • • i—ii —Ii —ii—i 34 O v o CO VO OH VO r VOO vN oO VN OV O 00 VO £N- O O NO NC Ov Oo N VO vo VO O hD ft § ■a ft co f ol vo o u u © ft P • * o o C vo CO O • CM • CM CM v oV O CO CO O VO V G O r0H 3 0 pcO U 0 C O ft e 0 © a lfteO Ss s; O §£ 123O a o O 3 4 O •H C O U rCH 0O cO ft • cO O H CM COft- vovO £>-00 hS) * P H CO 34 P •H •H bO P cO •rl •rl hfl 0 £ f 0t 0 •b D £ r l bfl 3 cO 0 J U £0 0‘ 3O4 5Tifft •» 34 0 0 •H ft c 0 u 0 P •H 0 O 3-4 p 34 •H P P ft 0 0 0 0 P 34 cO 0 O •rl O H •H 0 se so ■ H ft ft £5 3H 4 •c O 1 e-t ft fi •b0H £ ©t8II —1• f fOt 11 O i>4 34 ft p34 0 o § u u © 0 ft c 3 4 O O C0 MC CNO CO MC CO O S-. 0 -C • • • • i —IrH i —It—I o 4 •3 OH K'S 'ft • -P £ 15 33 ft ■ftON rH rH rl N t S O * ft ft ft 0 hO P ft vo 34 5 4 0 U f0t ■8*E 0 E •rl ft 3 ft r~ 0 • 34 O > <4 O O VO O ON on Q © •H O •rl •H f H t 0 ft7 -i 4 0 f H J -l0t•30 !Uf0t PO ft •H tcd? Gs* Pi to 0 • p 0 * -p O £ » | 0 CD **? C rH cd 0^ O C M rH qg OI »A 0 rH 3 5O C o G g i i—i P4 o» 3 o •H P cd 0 Pi CO • rH 4 ? — ® C rH cd 0 ^ S vrv 0 & rH rH 63 Os o CM ON O O H ci 00 PH • ON O OH • CM CM vr\ oS i—lO oI NO CM 00 I PQ • CM• CM CM IN e 00 O e I PQ C O rH • CM I PQ CM CO vpv I pq pq rH CM CM u"N PQ OH pe, £ £P cn- I ch £N- rH <3j k -3- O I CM Ei rH O O- OH rH 4 =4I O C 0) •H 0 ‘S •* rH O rH •r) P0J fi ?3. PCJ W O• CM M CO O •H CM CM O i —I CH I r® *8 i —i > © H On O a o NO o-. 00 '"t m S' i —I S' O O r H OI un O-3- CO o- | pq I PQ un UNqq (Protein consumed/body weight gain) pq ON I oNO Cvi a o s i < E o NO cqt On On ON ON I • o ON O N ON iH E -=*■ O I 00 CM ON ON to <0 * -P ■CD* £ Xi -P UN O C S CD C © O rH -H pn -P P. ct; co •sJ S C rH <0 J3 J UN 2 P0I 0 ,p -p CO • -p O C S 0 B c © o rH ‘H P, • Pt < oP jp Cd CO rH 0 > 0 § 0 rH UN level Protein efficiency S' a voH PQ 00 s o 3* Statistical results of Experiment 7: (Continued) 00 65 -3- \o r-j O N U On O fa 00 O I rH O n C\i O n £ CM PQ E co o 1 00 t>- I CO 00 o vo £ CM rH IN- I o i —i o 7 (concluded) Statistical results of Experiment PQ CM 82 O O CM O- CO ON ~ A o- o VO VO vo vo ON OV t r H <«J ON UN ON -3 - UN U N PQ ON I O ON INON P— CM I On VO PQ CM <—I er h • <4 'S ON VO CM CO VO VO CM On ON UN pq ON ON VO CM 0 a x0 s 10 C O -j82 * 3*=4 ON rH ON rH ON VO CM o •o 0 0 fa 0 0 0 M £ Sh 0 ^3 fa <> 3* UN • O 0 fa c S25 0 c o g E rH rH g > 0 h s> rH hO 0 rH •rH a a fa 0 2 ps CO 0 0 ^ a ^ 0 0 0 .£3 fa 00 0 • fa o S3 £3 0 E S3 0 O rH •H fa fa fa 0 3 PcJ (O £ rH b0 0 0 JQ r H 0 ^ UN 0 > 0 rH 66 When fish meal was added with corn oil and CFS (rations 8 vs 6), no effect on chick performance was observed* An improvement in weight gain as well as feed and protein effi­ ciencies was observed when terramycin was added to the ration contain­ ing corn oil (Rations 5 vs l)(P 0*05)# The addition of CFS or fish meal, singly or in combination, to the ration containing antibiotic (rations 2> 4 vs l) did not change chick performance, Reid et al. (1958) found the potency of UGF in dried distillers solubles decreases during storage. Since CFS and dried distillers solubles contain similar UGF (Experiment l), it is quite possible that the potency of CFS in present test decreased after prolonged storage. This probably accounts for the fact that the growth-promot­ ing effect of CFS observed at the fifth week disappeared by the end of the experiment* Combs et al* (1954) found that certain antibiotics spared the chick requirement for UGF* Therefore, the reduced potency of CFS in the present experiment was possibly masked by the- presence of the antibiotic* The ability of CFS to promote chick growth on a diet containing an antibiotic was demonstrated in Experiments 1 and 9, These results confirm the idea that the potency of this particular lot of CFS used in the present experiment was lowered due to prolonged storage. 67 e) Productive energy value of the basal diets EXPERIMENT 8 It was the purpose of this experiment to compare known sources of UGF and CFS, their ashes, and their sulfur contents on chick growth* Bay-old, male, White Plymouth Rock chicks from a commercial hatchery were distributed as described previously, with three repli­ cates of ten birds per treatment® The average initial weight per chick of each pen was U1 grams* The composition of the basal ration is presented in Table 18, Materials were ashed by charring and then placing in a muffle fur­ nace at 525° C* for approximately 13 hours* Their calcium, phos­ phate and sulfur contents were analyzed according to AQAC methods (1955) and are presented in Table 9* During charring and ashing certain amounts of sulfur might have been driven off* The sulfur content in the ashes therefore, were lower than that reported in Table 34-« Three types of sulfur were added at graded levels* The calculated protein, calcium and phosphate levels of all rations were equalized to that of the basal* Group weights were taken at the first day and the end of fourth week* The results are presented in Table 20* The statis­ tical results are indicated by asterisks since the usual method of underling (Duncan!8 test, 1955) would be too long for 19 treatments.. All treatments, except two higher levels of sodium sulfate, showed no effect on chick growth at the end of four weeks* These 68 TABLE 18 THE COMPOSITION OF BASAL RATION FOR EXPERIMENT 8 Ingredients Lbs/Cwt Ground yellow c o m 56*71 Dehulled soybean oil meal 36*45 Animal grease 3*oo Methionine 0.05 Ground limestone 1.31 Dicalcium phosphate 1.88 Iodized salt 0.50 Trace mineral (Delamix)* 0.10 Calculated values: Protein, Productive energy, Cal/lb Metabolizable energy, Cal/lb Vitamins 23.04 969 1227 Grams added to 100 lb ration Vitamin A(25Q,000 lU/gm) Vitamin Dn (1,500 ICU/gm) Alpha-tocopherol acetate (20,000 ICU/lb) Menadione Thiamine hydrochloride Calcium pantothenate Niacin Pyridoxine hydrochloride Choline chloride Folic acid Inositol Vitamin B-io /Sm ) Total * See Table 13 1.60 26.00 4.60 0.10 0.30 0.50 1.50 0.20 10.00 0.10 0.10 0.60 45*90 grams 69 * in ■ta o O CV • Si O vO • CM -S' • O 3 CM C - VO VOO tN- s —1 00 H r 0 0 <+H o -p G 0 o Fh CtJ O 00 O ON • co• ON 1 I — s PQ <4 EH rH £ O •H -C0TPJ Cm rH 2 0 ,WG S vn S' vO s• H oo * vn rH 00 VT\ w O'N* 0 -P CO U 6 0 -P M -p £ 0 0 c 0 0 8 0 0 U -P rH 1— ! •H 0 ■H 'Xj t3 0 & 0 0 5 .5 0 0 *H Pi Q 0 cd r0Cj CO 03 HN 0 5 -cP 0 cd 0 U u © -P 04 g 0 0 G O O x: 0 •H G oO c o % > r? S P co 70 results indicate that the basal ration did not meet the conditions for sources of UGF to exert their growth-promoting effect0 The addition of 0.013 and 0*022 per cent of sodium sulfate de­ pressed chick growth severely (P^O.Ol)., However, chick growth was not affected by the addition of the same or higher levels of sulfur, when supplied as flowers of sulfur or calcium sulfate* Therefore, it was sodium, rather them sulfate, which caused the growth-depression. Since the depressed growth rate, due to the addition of 0.013 per cent sodium sulfate, was lower (P^O.01) than 0.022 per cent, it indicates the possibility of an imbalanced relationship between sodium and potassium or other nutrients contained in the ration. Burns and co-workers (1953) found that both sodium and potas­ sium are toxic to chicks if one was fed in excess of the other, and this toxicity could be corrected by raising the level of the other element. This finding explains the results obtained in the present experiment. 71 TABLE 20 THE EFFECT OF KNOWN SOURCES OF 130FS AND CFS, THEIR ASHES AND SULFUR CONTENTS ON CHICK GROWTH TreatmentBasal Ash of fish concentrate Ash of CFS Ash of dried distillers solubles Ash of vigofac Flowers of""'sulfur Flowers of sulfur Flowers of sulfur Sodium sulfate Sodium sulfate Sodium sulfate Calcium sulfate Calcium sulfate Calcium sulfate Calcium sulfate Fish concentrate CFS Dried distillers solubles Vigofac Percent ration Equivalent intact Sulfur material supplied 30 2.8 3 .5 0 .1 4 0.001 0.003 0.001 0.005 0.001 0.003 0.005 0.001 0.003 0.005 0.001 0.003 0.005 0.010 30 2.8 3 .5 0 .1 4 ** Significant at 1$ level from all other groups. Av. 4-week weight, gm 362 372 355 363 360 366 350 365 369 130** 186** 370 374 372 356 384 359 352 371 72 EXPERIMENT 9 It was the purpose of this experiment to study the influence of the productive energy value of the basal ration and supplemental trace minerals on the growth-promoting effect of CFS# Two basal rations were formulated (Table 2l)s Basal Li This ration contained 2D per cent protein, 1.2 per cent calcium and 996 Calories of productive energy per pound of feed# Basal B : Protein and productive energy levels of this ration were raised up to 22 per cent and 1085 Calories per pound of ration, respectively* The calcium content was the same as in A 0 The C/P ratio, based on productive energy, was 50 for basal A and 49 for B* Ihen metabolizable energy values were used, all con­ tained the same C/P ratio of 62# Delamix, a commercial mineral supplement containing trace elements other than manganese (Table 13), was employed to replace the manganous sulfate, Solk-Floc and 0*07 per cent of ground lime­ stone in basal rations A and B* Therefore, the protein, calcium, manganese and energy levels were equalized to the respective basal ration# Three per cent of CFS was added to basal rations A and E, with or without supplemental trace elements# The calculated protein level of each experimental ration was equalized to its correspond­ ing basal ration# Individual weights were taken at the end of eight weeks# Ana- 73 TABLE 21 THE COMPOSITION OF BASAL RATION OF EXPERIMENT 9 Ingredients Ration A B Lbs/ Cwt Ground yellow c o m Dehulled soybean oil meal Animal yellow grease Ground limestone Dicalcium phosphate Iodized salt Manganous sulfate (70$) Solka-floc Vitamin and antibiotic mix 63.44 28.83 2.98 1.67 2.00 0.50 0.02 0.01 0.55 49 .46 35.30 10.49 1.67 2.00 0.50 0.02 0.01 0.55 Calculated values; Protein, $ Calcium, $ Productive energy, Cal/lb Metabolizable energy, Cal/lb C/P (productive) (metabolizable) 20 1*2 996 1249 22 1*2 1085 1371 50 62 ^9 62 Vitamin and antibiotic mix Vitamin A(10,000 IU/gm), lb Vitamin Do (3t000 ICU/gm) , lb Alpha-tocopherol acetate (20 gm/lb), gm Menadione (crystalline), gm Vitamin mix No. 1, lb Choline chloride (25^). lb Vitamin (6 mg/lb), lb Terramycin (10 mg/lb), lb Total Unit/100 lb ration 0*06 0.03 4*5 0.04 0*10 0*20 0.10 0*05 0*55 lb * Supplied: riboflavin 2, pantothenic acid 4, niacin 9» and choline 10 grams per pound. 74 lysis of variance showed that the treatments altered the final weights and feed efficiencies* The magnitude of the differences were highly significant* Although Basal A contained less protein and energy than Basal B, these differences did not affect final weights* However, feed efficiency was improved (P-«£^0*0l)* The substitution of Delamix for raanganous sulfate in Basal A. depressed growth rate (P-*£l0*05), but showed no effect on feed effi­ ciency* This substution in Basal B did not affect chick growth but improved feed efficiency (P-*c.0*05)* Since manganese is not the only one mineral in Delamix, the other trace elements must account for the effects observed* These data, therefore, show that the chick requirements for trace minerals, other than manganese, vary with the protein and energy contents of the ration* Although the incorporation of CFS in Basal A showed no effect when added to Basal B it increased final weights (P-<^0*0l)o These results emphasize that tte plane of nutrition— levels of protein and energy— influences chick response to CFS* CFS increased growth rate (P-*^ 0*05) when added to Basal A with supplemental trace minerals* This indicates the ability of CFS to correct the unbalanced mineral relationship created by the elements, other than manganese supplied by Delamix* The increased body weight obtained by adding CFS to Basal B with supplemental trace minerals was not statistically significant* However, the addition of CFS to Basal B improved chick growth (P«^0*0l)* let, the substitution 75 TABLE 22 THE EFFECT OF CFS ON CHICK PEEFQRMANCE AS INFLUENCED BT THE P QMP CSITIONS OF RATION (Experiment 9) Treatment Av. final w t . gm • Feed/gain A 1321 2.39 A + CFS 1314 2.32 A + trace minerals (Tm) 1253 2.30 A + CFS + Tm 1329 2.38 B 1323 2.19 B + CFS 1414 2.10 B + TM 1385 2.06 B + CFS + Tm 1428 2.03 * Four replicates of 10 birds each were used per treatment. The average initial -weight per bird in each replicate was approximately 40 grams. 76 oo cm dr rH to On cp • CM 00 cp CM TO CO 5 o c © CP <— t •H O ■H O to£ o cm cm CM rH • CM 0 S' E O CP 0 op CM TO CM O On -CP £ CP I—I CM cp ■8 <8 ■—I cp l —I 0 © B Em m VO O • CM &I 00 U *s co cp m-n CM r—I CP *o TO o CM rH 8© CO (D 0 > 0 rH .CQ CS ♦rH sn CCS t-* § 0 & rH i —I 0 t> 0 rH S' o •rH -P •H -P CCS -P CO 0 0 I—I ■8 •rH U 0 > 1— 1 0 > 0 1— 1 rH 0 > 0 i— 1 rH 77 of Delamix for manganous sulfate in Basal B did not change the chick growth rate* Therefore, the above results indicate clearly that CFS was able to improve chick body weights, with or without complex trace mineral supplement* The incorporation of CFS to the basal ration of Experiment 8, containing 23 per cent protein and 969 Calories of energy, did not stimulate chick growth* However, the growth-promoting effect of CFS was detected on ration containing 22 per cent protein and 1085 Calories of energy, in Experiment 9* Thus, these two experiments indicate that the energy level influences chick response to the UGF supplied by CFS* This influence was also illustrated in pre­ vious experiments (Table 23)* The question, whether it is the level of energy or fat which realy regulates chick response to CFS, naturally comes up at this point« The mechanism through which fat exerts its effect appears to differ for different sources* Rand et al* (1957) reported that on isocaloric diets corn oil improves chick weight gain and protein re­ tention* therefore, other than its caloric value, corn oil appears to possess an additional intrinsic value* Baldini and Rosenberg (1957), and Vondell and Ringrose (1958) found the increased energy level by adding beef tallow or raising carbohydrate content equally improved chick growth and feed efficiency* Thus, the nutritional effect of tallow, aside from its fatty acid content, is credited entirely to its caloric value* Gonsequantly, beef tallow does not contain the addi- 78 PS 0 •rt

H 2 0 o H *0 e 0,s 2 O rH VP» VO ON XO VO Ov I O v O 00 Ov • o• vO o « o• C^i -p s o I —I I Ov O o o 00 00 vrv xr\ l VO I i—I cu CM v r\ CM -b" O Ov vO ov Ov S' CM O- Ov XO CV OV VO Ov vrv O oo rH • 00 vO Ov • CM s 07 c0 <3} 3 «H 0 0 O ■o u 3 ft -P X fcO •H 0 ? i —1 *8 * 6 § •* 0 £ X -P O0 0 CO i—1 £hi00 0 0- • 0 sfr 0 C rH CO cNo X CO 1—1 iH r—1 S CO 1—1 - N C O jv3o CM O CO CO CO 1—1 v 1—1 rH •H 0 O rH VO, CM a § ■£ O O £ il £ !>J ■8 X o X Eh -p c 0 -p 0 0 • § F-r u a X0 ON CO W r>Hs P 0 -©P F-< 0 0 u 0 -p 0 0 £ X •P VO VO VO rH CO NO NO On NO NO <>- VO vO CM CO ON CM vO rH -0NO 02 VO VO vo £ X o O o i —I •H O £ CO 0 xCTJ *H F-« 0 > g (D S •H 0 CO VO VO • <— 1 i —I + xCO fr, U O O m VO Vh vO g O CO VVO eR. 0 Ff-.t ft 0 00 •ud 0 • H£ X 0 Q -P rH 0 •OH o rH ft 0 0 0 F-r -0P o X • H 0O rH 0 ft 0 E U •H 0 0 0 S E o E X o .a a d o CO CO + §5 o o o o + CO CO 8 + + £ o s o VO • i— 1 o U P o CO * t •H X 8k a o \D ON vO § vR CO o E vo vo co -3- CO rH o K 00 VO 00 CO CO Is YRYR vD VO vo 00 oco vo vo rH CO S s S o K o a .. CM CO CO CO S SO O CO CO CO a o so 'co sP.Nco sP* 5* Statistical results for body weight. Experiment 10 VO CO CO 6 o E o O vo vo m • IN. CO VO CO VO vo • • E O Ov CM VO Avo S-'v A• vo • i —Ii —I i—Ii—I E O 'VO ^cA-v f5« vo• i —Ii —I © © & xJ Vp O vO YCO A. © © £ © rH X> c0 U cO > o CM vO 10 •rH OCO VO rH -3VO CO i—I 00 CM CO rH (— I SS o o s O E O VO VO G CO © i—1 © s> © 1—1 i—1 © > © i—1 VY VO YR rH +? 00 10 © I—I XC3O -SHH c0 > 00 i —I CO I —I vo o CO I —I rH a cO © © > © i —1 VO 85 v£> o\ 8 5o o VO On VP•vVP•v I —1<—I CK o~> Statistical results for feed efficiency, Experiment 10 58 Ov • ♦ i—Ii—I o O o Ov -p C O CD 0 rH .aa s •rH cij > > C co 0 CD rH CD > 0 i—I 86 tlon of two levels of corn oil meal to the ration containing CFS depressed growth at fifth and eighth weeks (P^O.Ol)* When fish meal and corn oil meal were added to ration containing CFS, chick growth rates for the fifth and eighth weeks were reduced (P^'0,01 and P ^ 0 * 0 5 > respectively)* Data from Experiment 1 indicated that the corn and soybean oil meal type of ration contains enough UGF supplied by fish meal* However, this type of ration is not sufficiently supplied with the UGF contained in CFS or dried distillers solubles* Consequently, the replacement of CFS for fish meal should increase chick perform mance* The results obtained in the present experiment substantiate this* According to Draper (1944) corn oil meal is deficient in cys«teine, lysine and possibly glutamic acid* Hence the replacement of corn oil meal for fish meal, or the addition of the former to ration containing CFS reduced certain critical amino acids in the ration and resulted in an upset in the amino acids balance* caused chick growth to be depressed* This probably 87 D» Availability or vitamins contained in CFS Concentrate J 2 * EXPERIMENT 21 Composition of the basal ration, grossly deficient in vitamins, is presented in Table 26* Three levels of CFS were added at the expense of corn and Amisoy, an isolated soybean protein, so that each ration contained 23 per cent protein and three per cent added fat* The results are presented in Table 27* Statistical results indicate that six per cent CFS promoted chick growth over the basal for both breeds of birds (P<*0«01)* The incorporation of three per cent CFS increased chick weight for White Leghorn but not White Plymouth Rock, male.® ( P ^ “0*0l)* ^hese indicate a breed difference in the response to CFS* The addition of CFS also improved the feed efficiency (Table 25)® Among White plymouth Rock birds, when 1*5 per cent CFS were used the feed efficiency was slightly decreased* However, when three or six per cent CFS were used the feed efficiency was sharply increased* The improvement in feed efficiency for White Leghorn chicks increased with each higher levels of CFS* The vitamin contents of this basal ration were lower than the NRG recommendation (Table 28)* Since only 0*1 gram of manganous sulfate (70%) was added per 100 pounds of ration, the basal ration was also deficient in this mineral* Such deficiencies were also reflected by the poor performances of the basal groups* 88 TABLE 26 THE CCRPCSITION OF BASAL RATION FOR EXPERIMENT 11 Experiment No. Ingredients 11 12 Lbs/Cwt Vitamin and antibiotic mix Vitamin A (10,000 IU/gm) Vitamin Do (1,500 ICU/gm) Alpha-tocopherol acetate (20 gm/lb) Riboflavin Calcium pantothenate Niacin Pyridoxine hydrochloride Choline chloride (25/») Folic acid Biotin Vitamin B±2 (6 mg/lb) Para-aminobenzoic acid Terramycin (10 gm/lb) I Ground limestone Dicalcium phosphate Iodized salt Kang anou s sulfat e (70^ ) OuSO. -5H20 CoS03-7H20 Fecl^ *6H20 23. 1 j Mineral mix 71.25 19-30 3.00 0.05 4.20 2.20 • 1 Calculated protein level, $ 74.21 19.00 3.00 0.05 3.719 0*021 and 0*5 per cent levels* In one other treatment 0,2 per cent of calcium in the form of ground limestone was added to ration con*-* taining 0*3 per cent sodium sulfate. Since the basal ration cal~ culated 1*31 per cent calcium, this addition made a total of 1,51 per cent* The results are presented in Table 30o The analysis of variance shows that the treatment did not alter chick performance. These results are in accord with the findings in Experiment 5* Sodium sulfate, added up to 0*5 per cent of the diet, did not affect chick performance « 94 EXPKRIiiiENT 14 This expariment was designed to study further the effect of sup­ plemental sulfate on chick performance* In Experiments 5 and 13 sodium sulfate was used whereas in this experiment calcium sulfate (CaSO^HgQ) was employed* The calcium level of all rations were equalized* Composition of the basal iation is presented in Table 31. For each level of added calcium sulfate an equivalent amount of ground limestone was substracted and the amount of corn and soybean oil meal adjusted so that the calculated protein and calcium levels were equalized to the basal ration* The results presented in Table 32 indicate that supplemental sulfur, added in the form of calcium sulfate up to 0.12 per cent of the diet, did not affect chick growth* 95 TABLE 31 THE COMPOSITION OF BASAL RATION FOR EXPERIMENT 14 Ingredients Ground yellow corn Dehulled soybean oil meal Animal grease Methionine Gr ound lime stone Dicalcium phosphate Iodized salt Trace mineral (Delamix)* Vitamin and antibiotic mix Lbs/Cwt 56.19 36.44 3*oo 0.05 1.30 2.30 0.50 o.lo 0.12 Calculated values: Protein, % Productive energy, Cal/lb Metabolizable energy, Cal/lb Vitamin and antibiotic mix Vitamin A (250,000 IU/gm) Vitamin Do (1,500 ICU/gm) Alpha-tocopherol acetate (20,000 ICU/lb) Menadione (8 gm/lb) Thiamine hydrochloride Riboflavin Calcium pantothenate (32 gm/lb) Niacin Pyrid oxine hyd rochloride Choline chloride Folic acid Vitamin B ^2 (1 Sodium procaine penicillin (500 gm/kg) ★ S e e Table 12 for composition. 23.00 963 1227 gm/100 lb ration 1.60 26.00 4.60 2.84 0.30 0.20 5.00 1.50 0.20 10.80 0.10 0.60 1.00 96 TABLE 32 THE EFFECT OF SULFUR, SUPPLIED AS CALCIUM SULFATE OS CHICK GROWTH* (Experiment 14) Tr eatment ________________________ Ave rag e final -weight. gm ________________________ Basal 235 0.0*$ s 2*19 0.06$ S 231 0.08$ S 243 0.10$ S 232 0.12$ S 237 * Day-old, straight-run, W.P.R. chicks were subjected to 1—week standardization before the 2-week experiment. Ten birds per treatment, with an initial average weight of 75 grams per bird were used. TABLE 33 THE EFFECT OF FLOWERS OF SULFUR ON CHICK GROWTH* (Experiment 15) Treatment Average final weight, gm Feed/gain Basal 224 1.75 0 .05$ S 218 1.73 0.10$ S 231 1.69 0 .50$ S 209 1.81 1.00$ S 223 — 1 .50$ s 234 1.67 * Day-old, straight-run, W.P.R. chicks were subjected to 1-week standardization before the 2-week experiment. Ten binds per treatment, with an initial average weight of 72 grams per bird were used. 97 EXPERIMENT 15 This experiment was designed for the same purpose as Expriment 14* However, sulfur was added as flowers of sulfur in basal ration of Experiment 14* The results are presented in Table 33. Upon statistical analy*-* sis, differences in final weights due to treatments were found not to be significant* Gordon and Sizer (1955) found that the addition of 0*5 per cent sodium sulfate to a ration containing 0*07 per cent inorganic sulfur showed no effect on chick growth rate* However, after the removal of this 0*07 per cent sulfur, the supplemental suEfLate stimulated chick growth* Therefore, the failure to observe any effect on chick growth by adding different sulfur~compounds to basal rations employed in these experiments indicates that they already contained enough inorganic sulfur* After the completion of Experiment 15, certain feed ingredients were analyzed for total and inorganic sulfur contents* Peroxide bomb or fusion technic (Parr Instrument Co$ 1950) was employed to determine total sulfur content* The inorganic sulfur content was determined by autoclaving the sample with hydrochloric acid for eight hours* on steam bath. It was filtered and then evaporated to a small volume The sulfur was determined as barium sulfate* The results are presented in Table 34» It is noticeable that the sulfur contents of most of the material tested were rather high. Considering dehulled soybean oil meal and 93 TABLE 34 SULFUR CONTENT OF CERTAIN POULTRY FEED INGREDIENTS Material Antibiotic mixture^" CFS concentrate No. 3 Sulfur content. $ Inorganic Total 0.4k 0.762 0.983 Dehulled soybean oil meal 0.51 Delamix, a trace mineral supplement 0.27 Dried distillers solubles 0.42 Dried whey 0.20 Fish concentrate, Sea Horse 0.92 Fish meal, menhaden 0.93 Ground yellow corn 0.22 Vigofac 5.99 1 0 .592 0.52 5.62 2 A mixture of 10, 10 and 2 pounds of TM-10, Aurofac and P-50, respectively. 1956 Samples. 3 1957 samples. 99 ground yellow corn contain 0*87 and 0*20 per cent of methionine, and 0*68 and 0*15 per cent of cystine, respectively; and assuming no other organic source of sulfur is present, it is calculated that dehulled soybean oil meal and ground yellow corn contain 0*187 and 0*04-3 per cent of inorganic sulfur, respectively. Consequently, only these two ingredients at the levels used in the basal rations of Experiment. 5 and 13 supplied 0.13 per cent inorganic sulfur* Delamix and probably the antibiotic supplement supplied additional inorganic sulfur0 This supports the conclusion that the basal rations contained enough inorganic sulfur to meet the needs of the birds* The proceeding studies show that chicks can tolerate dietary sulfur up to 1*5 per cent without ill effect* 100 • Influence of urea on sulfur utilization by chi oks 0 EXPERIMENT 16 The supplementation with inorganic sulfur showed no effect on chick growth in previous studies* Baldini and Rosenberg (1955) reported that the chick requirement for methionine increases with increased productive energy of the ration* It may be possible that the increased energy also increases the requirement for sulfur in order to meet the accelerated need for sulfur-containing amino acids* The increased utilization of methionine hydroxy-analogue by the addition of urea was observed among chicks by Sullivan and Birds (1956)* Among ruminents, the addition of inorganic sulfur increases the utilization of urea (Hunt et al*, 1954; Abbert et al., 1955)* Therefore, it may also be possible for supplemental urea to increase the utilization of inorganic sulfur if the chicks* requirement for sulfur-containing amino acids is raised by high energy ration* Since the sulfur contents of certain known sources of UGF and CFS are rather high (Table 34) it was the purpose of Experiments 16 and 17 to learn if the addition of urea to ration, deficient in sulfur-containing amino acids and containing over 1000 Calories of productive energy per pound of ration, improves the utilization of supplemental sulfur* After the completion of Experiment 8, the birds were fed a starting mash (54S-2) for one week and then used in Experiment 16* They were distributed according to weight and nutritional background* Two replicates of ten birds each were used per treatment* The r- 106 acids*. However, the addition of methionine to the basal rations did not affect chick growth* Therefore, it indicates that the rations are actually not deficient in these amino acids* and the published values on nutrient contents of feedstuffs, employed for the compu~ tation, do not apply in every case. Consequently, under the experi** mental conditions chicks would not be expected to show benefits from the addition of flowers of sulfur, calcium sulfate or urea singly or in combinations* 107 Importance of .supplemental vitamin K in rations containing corn and, soybean oil meal as the only sources of protein* EXPERIMENT 18 This experiment was originally designed to study the effect of menhaden fish meal and CFS on chick performance * An outbreak of hemorrhagic disease was encountered during the third to fourth week with 29 per cent mortality* Diagnosis of this disease was made by the Veterinary Pathology Laboratory of Michigan State University. The composition of the basal ration, presented in Table 38, was similar to all other rations employed in this series of studies in that soybean oil meal and corn were the only protein sources« However, vitamin K was absent In the basal ration of present experi­ ment.' Therefore, it is quite possible that the lack of this vitamin was responsible for the outbreak of hemorrhagic disease* Similar rations were employed by Anderson et al* (1954.) and Frost et al© (1956) to demonstrate the nutritional hemorrhagic syndrome of chicks and the curative values of different forms of vitamin K, respectively© The syndrome observed in the present experiment was very similar to that described by Anderson et al* (l954-)« It should be noted that vitamin K was also not added in Experi­ ment 2, but no hemorrhagic syndrome was observed* The discrepancy was well explained by the findings of Almquist and Stokstad (1936)© Vitamin K was found to be transferred from dam to chicks through the egg yolks. Therefore, the body store of this vitamin was probably higher in the birds used in Experiment 2© Since chick growth rate 108 is not affected directly by vitamin K deficiency until the hemorrhagic condition becomes severe (Almquist and Stokstad, 1936) the data ob­ tained in Experiment 2 are still valid* However, the results of the present experiment must have affected and thus, cannot be used# 109 TABLE 38 THE COMPOSITION OF BASAL RATION FOR EXPERIMENT 18 Ingredients Lbs/Cwt Ground yellow corn 68.03 Dehulled soybean oil raeal 28.00 Ground limestone 1.3& Dicalcium phosphate 1.92 Iodized salt 0.50 Trace mineral (Delamix)* 0.10 Vitamin mix 0.11 Vitamin mix Vitamin A (10,000 IU/gm) gm/lOO lb ration 20.00 Vitamin D^ (3*000 ICU/gm) ^.50 Riboflavin 0.09 Calcium pantothenate 0.20 Niacin 0 Choline chloride * See Table 13 . 25.70 110 Ho Other experiments EXPSRIMijNr 19 The experiment was designed to compare the effects of CFS and fish meal on chick performance* house #500. The birds were reared in poultry After the completion of this house a performance test was made which indicated the existance of differences due to position of batteries in the starter-room* In the present experiment lots were randomly assigned to various batteries. At the end of this experiment, it was found that difference between batteries were highly significant. used* Therefore, the data for treatments were not Ill EXPERIMEI\fT 20 This experiment was designed to study the effect of CFS and dried distillers solubles, with and without the addition of methionine and/or glycine to a high energy ration (productive energy » 1234 Cal/lb)* Gage differences: were found highly significant at the age of & weeks (growing batteries)* Therefore, no interpretation is given to the results of the present experiment* In this series of studies differences due to battery positions in the rooms of poultry house #500 were found in two out of the six experiments* Certain remedies, eg. the installation of space heaters, should be made to avoid the recurrence* 112 GENERAL DISCUSSION The basal ration of Experiment 1 contained all known nutrients at, or above, the levels required by chicks for optimum growth* When CFS was incorporated in this ration, keeping the protein and metabo­ lizable energy contents equalized, chick growth was significantly improved* ^his clearly indicates that CFS contains unknown growth factor(s) and belongs in that relatively small but important list of crude sources of UGF* Like other crude UGF sources, the growth-promoting effect of CFS was modified by certain factors* It is shown in these experiments that two dietary factors Influence chick response to supplemental CFS— -energy level and nature of fat (Table 23)* CFS stimulated growth in rations containing crude corn oil or animal yellow grease when the percentage protein, Calories of productive energy and C/P ratio, respectively, were as follows: 23*1011:44 (Experiment l), 23:1009:44 (Experiment 7) and 22:1085:49 (Experiment 9)« It did not stimulate growth when these values were: 22:935:43 (Experiment 7), 23:969:42 (Experiment 8) and 20:996:50 (Experiment 9) or when the rations contained refined cottonseed oil (Experiments J+9 5 and 6) regardless of the dietary caloric values„ Therefore, the caloric value of a diet rather than its protein level or C/P ratio regulates chick response to CFS supplementation* A productive energy value of over 1000 Calories per pound of feed thus appears to be necessary for CFS to exert its best in promoting chick growth* Crude corn oil or yellow grease can te used to enhance the energyvaiue of the ration* 113 The energy requirement of CFS appears to be an asset* As the energy values were increased and the C/P ratios kept about the same, the feed efficiency improved (Experiments 7 and 9)* The combination of CFS and high energy rations stimulated chick growth and improved feed efficiency, respectively, This synergistic effect resulted a broiler which weighed more than three pounds at eight weeks of age on slightly more than two pounds of feed per pound of gain (Experi­ ments; 1 and 9)« This efficiency is higher than that obtained by com­ mercial broiler producers in present-day practice« The rations used in this series of experiments were simple and easy to mix. They needed no supplemental methionine and required no further supplementation with a *'fish factor*1* Rations of Experiment 9 were specially formulated with commercial application in mind* Many factors are considered in formulating commercial rations*^— C/P ratio, sources of UGF, supplemental antibiotics, feed additives, the availability and digestibility of the nutrients, etc* The abi­ lity of GFS to promote chick growth on diets containing an antibiotic was well demonstrated in Experiments 1 and 9* not limited to its UGF only* The value of CFS is Its protein, vitamins and energy values were also utilized by the chicks (Experiments 8, 9, 11 and 12),In case the energy value of a ration falls short of 1000 Calories,the addition of CFS would raise the caloric value as well as supply UGF, of CFS is distinct from its caloric value* The UGF content For example in Experiment 1 when caloric values were equalized, CFS stimulated chick growth* 114 Although CFS and certain other sources of UGF contain substan­ tial amount of inorganic sulfur (Table 34*), up to 1.5 per cent of the latter in the diet did not affect chick performance. Thus, the sulfur content was not responsible for the improved growth rate of the chicks>* The wet-milling industry produces CFS year round at a rather uniform rate* This is in contrast to certain other sources of UGF which are produced seasonally* This uniform rate of production of CFS should be of considerable importance to the feed industry* 315 coiCLuaious Twenty experiments, involving over five thousand chicks, were designed and carried out to evaluate nutritionally corn fermentation solubles (CFS Concentrate #3) and compare it with recognized sources of unidentified growth factors# Under the experimental conditions it was found that: CFS Concentrate #3 supplied unidentified growth factors required by chicks as well as protein, known vitamins and energy# Prolonged storage of the product reduced its growth-promoting value# CFS Concentrate #3 replaced equivalent amounts of fish meal in the particular basal ration used# CFS Concentrate #3 exerted its best effect in stimulating growth of meat-type chick® in diets containing over 1000 Calories of pro­ ductive energy per pound of feed# It was effective in the presence of dietary antibiotic# White Leghorn male birds were more responsive to the supple­ mental CFS Concentrate #3 than straight-run chicks of the same breed# A decrease in the growth rate of White Leghorn male chicks due to the addition of six per cent of refined cottonseed oil was restored by the addition of CFS Concentrate #3* Based on the growth data, it appeared that CFS Concentrate #3 may contain the same or similar unidentified growth factor(s) as dried distillers solubles# 116 LITERATURE CIT£B Ackerson, C. W., W. E. Ham and F. E. Mussehl, 1940* ■‘■'he utilization of feed elements by growing chicks* IX. The nitrogen of urea* Nab* Agr. Exp* Sta* Bui. lgOsl-7* Albert, 1|. W., U. S. Garrgus, R. M. Forbes and'W. H. Hale, 1955* Modified urea supplements with corn silage for wintering ewe lambs. J. Ani. Sci. 14:143-152* Almquist, H. J., and E. L* R. Stokstad, 1936* Factors influencing the incidence of dietary hemorrhagic disease in chicks* J. Nutr. 12:329^335* Anderson, G. C., J. H. Hare, J. K* Bletner, C. E. Weakley, Jr., and J. A. Mason, 1954* 4 hemorrhagic condition in chicks fed sim~ plified rations. Poultry Sci. 33:120*126* Arscott, G. H., and G* F. Combs, 1955* Unidentified growth factors required by chicks and poults* 4* Experimental variables which influence the dietary requirements of chicks for these factors* Poultry Sci. 34:843-850« Association of Official Agricultural Chemists, analysis, 1955. Official methods of Baldini, J. T., and H. R. Rosenberg, 1957* The effect of calorie source In a chick diet on g r o w t h , feed utilization, and body composition«, Poultry Sci* 36:432-435* Barnett, B. D., and H. R. Bird, 1955. Standardization of assay for unidentified growth factors* .Poultry Sci. 34:1178* Barnett, B. D. and H. R. Bird, 1956* Standardization of assay for unidentified growth factors* Poultry Sci* 35:705-710. Bice, C. M., and L. A. Dean, 1942. Utilization of urea nitrogen by growing chicks. Poultry Sci. 21:15-17* Bosshardt, D* K., J. W* Huff and R. H. Barnes, 1956* Effect of bromine on chick growth. Proc. Soc. Expt* Biol. & Med. 92: 219- 221* Brautlecht, C. A., 1953. Starch. Its sources, production and uses* Reinhold Publ. Corp. New York, N.Y. Briggs, G* M., 1956* Inadequacy of certain salt mixtures used in studies of unidentified growth factors for chicks. Poultry Sci* 35:740—742* 117 Briggs, G. M., G. F • Combs, L. Friedman, J. C. Fritz, R. J. Lillie, J. W. Nelson, F. W. Quackenbush and H. W. Titus, 1956* The ANRC reference chick diet. Feed Age. May 1956, pp. 41-44* Burns, C. H., W. W. Cravens and P. H. Phillips, 1953* The sodium and potassium requirements of the chick and their interrelaticn •* ship* J. Nutr* 50s317-329* Camp, A. A., H. T. Cartrite, J. H. Quisenberry and J. R. Couch, 1955* Further information concerning unidentified chick growth factors Poultry Sci. 34s559-566* Camp, A. A*, B* L. Reid and J. R. Couch, 1956* Growth promoting activity of ash when fed in practical diets to chicks* Poultry Sci* ,25:621-626* Camp, A. A., H. T. Cartrite, B. L. Reid, J. H. Quisenberry and J. R. Couch, 1957* Corn steepwater solubles as a source of unknown growth factor(s) for growing chicks* Poultry Sci. 36:13541959. Carlson, C. W., R. F. Miller, II. T. Peeler, L. C. Norris and G. F. Heuser, 1949* The complex nature of the animal protein factor* Poultry Sci. 28:750-752* Carlson, C. W., and E. Leitis, 1957* Methionine, betaine and choline as counteractants of selenium toxicity. Poultry Sci. 36:1108* Combs, G. F., G. H. Arscott and H. L. Jones, 1954* Unidentified growth factors required by chicks and poults. 3* Chick studies involving practical-type rations* Poultry Sci. 33:71-79* Combs, G. F., G. B. Sweet, H. L. Jones, G. L. Romoser and R. W. Bishop, 1954* Multiplicity of unidentified growth factors re­ quired by chicks and poults. Poultry Sci. 33:1050* Couch, J. R., J. R* Reed, Jr0, B. L. Reid and J. Ti7. Dieckert, 1952* Distillers solubles and other sources of unidentified factors for promoting the growth of chicks and poults* Proc. Seventh Distillers Feed Conf* pp. 59-71* Couch, J. R., J* R. Reed, Jr., R. L. Atkinson, R. W. Perrett, B. C. Welch and J. W. Dieckert, 1954* Fl*oc« Ninth Distillers Feed Conf* pp* 38-48* Couch J. R., B. L. Reid, A. A. Camp, T?. N. Dannenburg and E* E. Fiozacky, 1955* Distillers dried solubles in the diets of chickso Proc. Tenth Distillers Feed Conf. pp. 47-57* 118 Dannenburg, W. IT., B. L. Reid, 3. E. Rozacky and J. R. Couch, 1955. An inorganic chick growth response. Poultry Sci. 1023-1026* Davidson, J.A., W. N. Fischer and H„ C. Zindel, 1957. Michigan broiler test No. 4> Fall 1956* Michigan Agr. Exot* Sta. Quarterly Bui. 22:594-598. Davis, R. L., G. M. Briggs and H. J. Sloan, 1953* Effect of cobalt in diet of the chick* Proc. Soc. Expt* Biol. & Med. 82:175 Donaldson, F. E., G* F. Combs, and G. L. Romoser, 1956. Studies on energy levels in poultry rations. 1* The effect of calorieprotein ratio of the ration on growth, nutrient utilization and body composition of chicks. Poultry Sci. 35:1100-1105. Draper, G. I., 1944* 1he nutritive value of corn oil meal and feather protein. Iowa Agr. Exp. Sta. Res. Bui. 326. J Duncan, D. B., 1955* 11:1-42.^ Multiole range and multiole F test. Biomet. Fisher, H*, H. M. Scott and R. G. Hansen, 1954* Further studies on the alfalfa factor and its relation to the liver and whey factors. J. Nutr. 52:13-24. Fraps, G. S*, 1946* Composition and productive energy of poultry feeds and rations* Texas Agr. Expt. Sta. Bul0 678. Frost, D. V., H. S. Perdue, and H. C. Spruth, 1956. Vitamin K activity of menadione sodium bisulfite in chickens. J* Nutr. 59:181-196. Fuller, H. L., C. F. Carrick and S. M. Hauge, 1952. A compzrison of Bhp, fish solubles and whey in the growth of chicks. Poultry Sci. 21s473-478. Gordon, R* S., and I- W. Sizer, 1955* stimulate growth of the chicken. Ability of sodium sulfate to Science 122:1270. Hale, F. H*, and TJ„ S.. Garrigus, 1953* Synthesis of cystine in wool from elemental sulfur and sulfate sulfur. J. Ani. Sci* 12: 492—496. Higgins, W. S., D. A. Richert and *W F. Westerfeld, 1956. Molybdenum deficiency and tungstate unhibition studieso J. Nutr. 59:539559* Hill, C. H., R. L. Borchers, C. W. Ackerson and F* E. Mussehl, 1953* Studies on chick growth stimulation by alfalfa juice, casein and aureomycin. Poultry Sci. 222775-779o 119 Hill, F. b1., and D* L. Anderson, 1955* Comparison of productive energy and metabolizable energy determinations with chicks« Poultry Sci* 34:1201. Hill, E. G*, and G. M. Briggs, 1950* Studies on an unidentified growth promoting factor in cereals and other natural products for chicks* Poultry Sci* 29;763* Hill, F. W., and L. M*. Dansky, 1950* Studies of the protein require­ ments of chick and its relation to dietary energy level* Poultry Sci* 22*763* Hill, F. W*, and Ruth Renner, 1957* Metabolizable energy values of feedstuffs for poultry and their use in formulation of rations* Proc, 1957 Cornell Nutrition Conf* pp* 22-32* Hill, F* W*, D. L. Anderson and L. M. Dansky, 1954* The relation of dietary energy level to rate and efficiency of egg production* Proc*, 1954 Cornell Nutrition Conf* pp* 47-52* Hill, F. W., and L. M» Dansky, 1954* Studies of the energy require­ ments of chickens. 1* The effect of dietary energy level on growth and feed consumption. Poultry Sci. 33:112-119* Hubbell, C. H., 1957. Feedstuffs analysis table. Feedstuffs* Hunt, C. H., 0. G. Bentley, T. W. Hershberger, and J. H. Cline, 1954* The effect of carbohydrates and sulfur on B-vitamins synthesis, cellulose digestion, and urea utilization by rumen microorganisms in vitro* J. Ani* Sci. 13:570-590* Johnson, E. L., 1953* Season and carbohydrate source affect chick response to unidentified factors* Poultry Sci; 32:930-931* Herr, R. tr*, 1950* Chemistry and industry of starch. Academic press* Inc*, New York, N.Y. X Kohler, G„ P., and W. R. Graham, Jr., 1952* The seasonal response of chicks to an unidentified growth faegor found in fourage juice. Poultry Sci. 31s284-286* Kurnick, A. A., B. L* Reid, R. N. Burroughs, H. D. Stelzner and J. R* Couch, 1957. Effect of distillers dried solubles and molybdenum on the growing chick. Proc, iSco* Expt. Biol, & Me d. '25*353-356. Leach, R. M*, and L. C. Norris, 1957. Studies on factors affecting the response of chicks to molybdenum. Poultry Sci* 3 6 :1136* 120 leach, R. M. , Jr., and L. G. Norris, 1958* Recent research on the potassium requirement of chicks and turkey poults* Proc* 1958 Cornell Nutrition Conf* pp, 54-59* Leong, K. C., M. L. Sunde, H. R. Bird and C* A. Elvehjem, 1955. Effect of energysprotein ratio on growth rate, efficiency, feathering, and fat deposition in dickens, ^oultry Sci* 34: 1026-1027. Liggett, R. T?., and H. Koffler, 1958* Corn steep liquor in micro­ biology* Bacteriol* Rev*. 12:297-391. Lillie, R* AT*, J. R* Sizemore and H* R. Bird, 1953© Unidentified factors in poultry nutrition* 1, Development of chick assay. Poultry Sci* 32:855-862* Lillie, R* J., H* Menge, J, J. Miner and C. A* Denton, 1958* A breed difference with respect to carry-over effect of unidenti­ fied growth factors* Poultry Sci. 37:344-352* Lowe, I, P., and K. Roberts, 1955* Incorporation of radioactive sulfate sulfur into taurine and other substances in the chick embryo* J. Biol* Chem. 212:477-483* Machlin, L* J*, 1955* Studies on the growth response in the chick from the addition of sulfate to a low-sulfru diet* Poultry Sci. ^:1209* Machlin, L. J*, C. A. Denton, F. L. Kellogg and H. R. Bird, 1952* Effect of dietary antibiotic upon feed efficiency and protein requirement of growing chickens. Poultry Sci. 31:106-109* Machlin, L. J.,C* A. Denton, P. B. Pearson and H* R. Bird, 1952. Synthesis of cystine from inorganic sulfate by the hen. Fed, Proc* 11:448-449. Machlin, L. J,, P. B. Pearson, C. A. Denton and H. R. Bird, 1953* The tilization of sulfate sulfur by the laying hen and its incorporation into cysine* J. Biol* Chem, 205:213-219. Machlin, L. J., J* R* Jackson, A. H. Lankenau and P. B. Pearson, 1954* Uptake of feathers, gizzard lining, liver and muscle of young chickens after injection with radioactive methio­ nine or sulfate* Poultry Sci. 33:234-238* Machlin, L. J., P. B. Pearson and C. A. Denton, 1955* ^he utiliza­ tion for the synthesis of taurine in the developing chick em­ bryo* J* Biol. Chem. 212:469-475* 121 March, B. E., and J, Biely, 1954-• The nutritive value of fats of different origin in chick starters* Poultry Sci. ,22:1069. March, B. E., and J* Biely, 1955* Fat studies in poultry. 3. Folic acid and fat tolerance in the chick. Poultry Sci. 34:39-44* March, B. B*, J* Biely and S. P. Touchburn, 1955* Studies on an unidentified chick growth factor in dehydrated green feed. Poultry Sci. 34.:968-972. McKittrick, B. S., 194-7* The interrelations of choline and methionine in growth and the action of betaine in replacing them. Arch. Biochem. 15:133-155. Menge, H., G. F. Combs, Peng-tung Hsu and Mary S. Shorb, 1952. Uniden­ tified growth factors required by chicks and poults. 1. Studies with chicks using purified diets* Poultry Sci. 31:237-24.7* Mitchell, H. H., and T. S. Hamilton, 1929* The biochemistry of the amino acids. Chem* Cat. Co., Inc., New'*’York, Am. chem* Soc* Monograph Series: 619* Morrison, A. B., 1955. ^he present status of unidentified growth factors in poultry hutrition* Proc* 1955 Cornell Nutrition C onf. pp * 72-79* 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* Morrison, A. B*, R. Dan, L. C. Norris and M. L. Scott, 1956* Further evidence on the requirement of the chick for unidentified minerals. J. Nutr* 60:283-295. Morrison, A. B*, L* C. Norris and M. L. Scott, 1956. Studies on isolation of the chick growth promoting factor(s) in the ash of unidentified growth factor supplements* Poultry. 35:1160-1161* Morrison, F. B., Feeds and Feeding, 1950* N. Y. Morrison Publ* Co., Ithacal Moyer, A. J.,and R. D. Coghill, 194-6. Penicillin VIII* Production of penicillin in suface cultures. J. Bacteriol. 51:57-78. National Research Council, 1954. Pub* 301* Nutrient Requirements for Poultry Nesheim, M. C., and M. L. Scott, 1957. Nutritive activities of sele­ nium for chicks and poults. Proc. 1957 Cornell Nutrition Conf* pp* 48-52* 122 Norris, L # C*, 1954* dried solubles* Unidentified chick growth factors in distillers Proc* Ninth Distillers Feed Conf* pp. 20-28. Norris, L. C., 1955* decent progress on unidentified growth factors in distillers dried solubles and other feed supplements* Proc. Tenth Distillers Feed Conf* pp. 39-42* Norris, L. C*, and T. R. Zeigler, 1958® ments of chicks and turkep poults* Conf. pp, 72-78. Studies on the zinc require­ Proc* 1958 Cornell Nutrition Norris, L. C., R. M* Leach, Jr., and T. R* Zeigler, 1957* Requie— ments of poultry for molybdenum, zinc and other trace elements. Proc* 1957 Cornell Nutrition Conf* pp. 33-47» Norris, L. C*, R. M. Leach, Jr., and T. R. Zeigler* Recent resaerch on the mineral requirements of poultry* Proc* 13th Distillers Feed Confer* pp. 63—73* O'Dell, B* L*, M* D* Boston, J* E* Savage and A. G. Hogan, 1952* The use of isolated soybean protein in synthetic diets for chicles. Poultry Sci* ^1$559-563. O'Dell, B. L*,snd J* E. Savage, 1957, Potassium, zinc and distillers dried solubles as supplements to a purified diet* Poultry Sci. 36:459-460* Parr Instrument co*, Moline, 111,, Direction Booklet No. 116* Patterson, E* L*, R* Milstrey and E. L. R* Stokstad, 1957* Effect of selenium in proventing exudative diathesis in chicks* Proc, Soc* Expt* Biol* & Med* 95s617-620. Peterson, D. W#, 1950a. Some protperties of a factor In alfalfa meal causing degression of growth In chicks* J* Biol* Chem* 183:647— 653* Peterson, D* 17*, 1950b. Effect of sterols on the growth of chicks fed high alfalfa diets or a diet containing quillaza saponin* J* Nutr* 42:597-607. Petersen, C. F., A. C* Wiese and A. R* Pappenhagen, 1955. Unidenti­ fied chick growth factors. 1* Purified assay diet and crude supplement response* Poultry Sci* 34s673-678* Philips, A. G*, R. H. Carr and D. C. Kennard, 1921* Green feed versus antiseptics as a preventive of Intestinal disorder of growing chicks. J. Agr. Res. 20:869-873. 123 Hadley, J. A#, 1954* Starch and its derivatives. Wiley & Sons Inc** New York, N.Y. Vol. II. John Hand, IT. T., H. M. Scott and F. A. Kumraerow, 1957. The utilisation of fat by the growing chick. Poultry Sci. 36:1151-1152. Rasmussen, R. A., P. W. Luthy, J. M* Van Lanen and G. S. Borruff, 1954. The dual nature of the unidentified chick growth promoting activity of distillers dried solubles. Proc. ITinth Dis­ tillers Feed Conf. pp. 29-37. Rasmussen, R. A., P. Y". Luthy, J. M. Van Lanene and C. S. Borruff, 1957. Measurement and differentiation of unidentified chick growth factors using a new semipurified ration. Poultry Sci. ,26:46- 54* Reid, B. L., A. A. Kurnick, R. L. Svacha and J. R, Couch, 1956., The effect of molybdenum on chick and poult growth. Proc. Soc. Expt* Biol. 8c Med. 93:245-248. Reid, B* L., R. L. Svacha, A. A. Kurnick, F. M. Salama and J. R. 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