1+ Agriculture Canada Research Direction generale Branch de la recherche Technical Bulletin 1 986-4E The T.M.E. system of feed evaluation: methodology, feed composition data and bibliography The map on the cover has dots representing Agriculture Canada research establishments. ONE HUNDRED YEARS OF PROGRESS The year 1986 is the centennial of the Research Branch, Agriculture Canada. On 2 June 1886, The Experimental Farm Station Act received Royal Assent. The passage of this legislation marked the creation of the first five experimental farms located at Nappan, Nova Scotia; Ottawa, Ontario; Brandon, Manitoba; Indian Head, Saskatchewan (then called the North- West Territories); and Agassiz, British Columbia. From this beginning has grown the current sys- tem of over forty research establishments that stretch from St. John's West, Newfoundland, to Saanichton, British Columbia. The original experimental farms were established to serve the farming community and assist the Canadian agricultural industry during its early development. Today, the Research Branch con- tinues to search for new technology that will ensure the development and maintenance of a com- petitive agri-food industry. Research programs focus on soil management, crop and animal productivity, protection and re- source utilization, biotechnology, and food processing and quality. The T.M.E. system of feed evaluation: methodology, feed composition data and bibliography I. R. SIBBALD Animal Research Centre Ottawa, Ontario Animal Research Centre Contribution 85-19 Research Branch Agriculture Canada 1986 Copies of this publication are available from: Director Animal Research Centre Research Branch, Agriculture Canada Ottawa, Ontario K1A 0C6 Produced by Research Program Service ©Minister of Supply and Services Canada 1986 Cat. No. A54-8/1986-4E ISBN 0-662- 14628-X - 1 - SUMMARY The true metabolizable energy (TME) system of feed evaluation comprises assays for bioavailable energy, amino acids, lipids and minerals. In this Bulletin the principles of the system are explained in the 'Introduction' which includes a diagram showing the partition of energy within the bird. A section entitled 'Bioassay Procedures' describes the basic methodology while sub-sections contain: a list of precautions to be observed when using the assays; a detailed description of the precision- feeding technique; details of the design and construction of cages for adult cockerels; and a discussion of excreta collection techniques. The section on ' Feedingstuf f Composition* contains data describing the nitrogen, ether extract, crude fibre, neutral detergent fibre, ash, calcium, phosphorus, gross energy, TME, TME corrected to zero nitrogen balance (TME ), total amino acids, and true available n amino acids (TAAA) of an array of feed ingredients; the data are expressed per unit of dry matter. An important component of the Bulletin is the 'Bibliography' which lists references to more than 500 publications concerned, directly or indirectly, with the derivation and application of TME- type data. RESUME La methode de l'energie metabolisable vraie (EMV) pour evaluer les aliments des animaux comporte des dosages biologiques de l'energie, des acides amines, des lipides et des mineraux biodisponibles . Dans ce bulletin, on explique d'abord les principes du systeme (Introduction) a l'aide d'un diagramme montrant la repartition de l'energie dans le corps de la volaille. Une section intitulee "Methodes de dosage biologique" decrit les techniques de base et comprend des sous- sections couvrant diverses questions comme: les precautions a prendre dans la conduite des dosages, une description detaillee de la technique d* alimentation de precision, les details de la conception et de la construction de cages pour les coqs adultes et une analyse de la technique de collecte des excrements. La partie sur la "Composition des aliments" contient des donnees sur les teneurs en azote, extrait ethere, cellulose brute, fibre au detergent neutre, cendres, calcium, phosphore, energie brute, fiMV, KMV ramene au bilan azote zero (EMV ), acides amines totaux et acides amines disponibles vrais (AADV) , de tout un assortiment de matieres alimentaires . Les donnees sont calculees sur la matiere seche. Complement important du bulletin, la "Bibliographie" contient plus de 500 titres de publications touchant de pres ou de loin a l'obtention et a 1 ' utilisation des donnees basees sur l'energie metabolisable vraie. - 3 - PREFACE In 1981 the Animal Research Centre published Technical Bulletin No. 3 entitled: Bioassays Based on Precision Feeding of Poultry. The French version was entitled: Dosage biologique en cage metabolique chez la volaille. The Bulletin contained a brief description of the precision feeding technique and of the assays based thereon. The primary purpose of the Bulletin was to be a bibliography of relevant literature and 162 references were listed. An enlarged and updated version entitled: The T.M.E. System of Feed Evaluation was published in 1983 as Contribution 1983- 20E of the Research Branch of Agriculture Canada. The French edition was Contribution 1983-20F and was entitled: La methode devaluation des aliments par le calcul de l'energie metabolisable vraie (E.M.V.). Features of the 1983 version were an enlarged section on assay methodology, inclusion of a section describing feedingstuff composition, and extension of the bibliography to 314 references. Demand for the revised Bulletin exceeded all expectations and approximately 1200 copies were distributed in less than two years. The present Bulletin is an updated version of the 1983 edition. The introduction and methods sections have been revised in the light of recent research. The feedingstuff composition section is greatly expanded by data describing additional samples and by the inclusion of total and true available amino acid values. More than two hundred references are added to the bibliography. Digitized by the Internet Archive in 2013 http://archive.org/details/tmesystemoffeede19864sibb - 5 - TABLE OF CONTENTS Suirana ry / Re s ume Preface Table of Contents Acknowledgements Introduction Bioassay Procedures General Precautions Precision Feeding Cage Design and Construction Excreta Collection Feedingstuff Composition Table 1 Table 2 Table 3 Table 4 Bibliography 1917 1924 1926 1966 1970 1975 19/6 197 7 1978 1979 1980 1981 1982 1983 1984 1985 Nitrogen, fat, fibre, mineral (g/kg) and energy (MJ/kg) concentrations in the dry matter of several feedingstuff s Total amino acids in the dry matter of several feedingstuff s (g/kg) True available amino acids in the dry matter of several feedingstuff s (g/kg) Bioavailability of amino acids in the dry matter of several feedingstuf fs (%) 1 3 5 6 7 12 12 16 17 22 28 34 37 52 59 66 73 73 73 73 73 73 73 74 74 76 78 80 86 92 99 105 112 _ 6 - ACKNOWLEDGEMENTS The author wishes to thank all those who helped with the preparation of this Bulletin. Several people provided references to TME-related publications while others provided samples of feedingstuff s for analysis; their cooperation is appreciated. The preliminary drawings for the figures illustrating cage design and construction were prepared by F. Nachbaur; final drawings and all photographs were prepared by Research Program Service, Agriculture Canada. The feed composition tables could not have been compiled without the technical help of S. Winter, R. Cochrane and L. Hope; the proximate and amino acid analyses were made, in part, by the Chemistry and Biology Research Institute, Agriculture Canada. The staff of the poultry unit of the Animal Research Centre, Research Farm provided valuable help by caring for birds and participating in bioassay work. Finally, my thanks to D. Leger for editorial assistance throughout the preparation of the Bulletin. DISCLAIMER Mention of a trade name, proprietary product, or specific equipment does not imply its approval to the exclusion of other products which may be suitable. - 7 - INTRODUCTION The true metabolizable energy (TME) system of feed evaluation, which is gaining wide acceptance (Figure 1), comprises bioassays for TME, THE corrected to zero nitrogen balance (TME ), true available amino acids (TAAA) , true available lipids (TAD and true available minerals (TAM) . Each assay involves corrections for metabolic plus endogenous losses. The desirability of making such corrections was recognized by Armsby (1)* who wrote: "since the digestion experiment as ordinarily conducted ignores the presence in the faeces of excretary products, the results obtained by its use will necessarily be too low." Goodwin's translation of Kellner (3) states: "the assumption is made that dung consists only of undigested food. The assumption, however, is not quite correct, for the faeces are always mixed with small quantities of substances which come from the digestive organs and which are termed the products of metabolism." Despite this early recognition of the problems associated with metabolic and endogenous losses, little attention was given to the introduction of appropriate corrections. The assay for the 'biological value' of protein included corrections for metabolic fecal and endogenous urinary nitrogen losses (2) and nitrogen-free diets have been used in some amino acid availability studies. However, the need for corrections has generally been ignored and the teachings of Kellner and Armsby seem to have been forgotten. By making corrections for metabolic and endogenous losses, the assays of the TME system should provide data which are independent of the species, strain, sex, age and physiological state of the assay bird. Ideally, the data should be independent of species in general but this state is unattainable because of the wide variation in microbial activity within the alimentary tracts of diverse species such as the cow, pig and chicken. Such microbial activity has profound effects on the composition of ingesta offered for digestion and absorption by the host animal. * Numbers in brackets refer to publications listed in the bibliography which starts on page 73. - 9 The term TME was defined by Harris (4). The effect of metabolic fecal and endogenous urinary energy losses on apparent metabolizable energy (AME) values was described, with theoretical data, by Guillaume and Summers (5). The relationships between AME and TME are described in detail in two recent publications (526,556). Corrections for metabolic and endogenous losses are equally important and have similar effects in TAAA, TAL and TAM assays. It is now common practice to correct TME values to zero nitrogen balance. The reasons are explained in detail by Wolynetz and Sibbald (526) who conclude that the correction causes estimates of bioavailable energy "to be based on the assumption that the available energy is being used for a single common purpose, the production of heat, and that the composition of the bird is unchanged." There are no comparable corrections in the TAAA, TAL or TAM bioassays. However, the TAM assay differs from other assays of the TME system because an intake of a mineral in excess of the requirement and storage capacity is likely to be excreted. Therefore in the TAM assay the mineral intake must be kept below the requirement. Intakes in excess of the requirement have no effect on the TAAA bioassay because the surpluses are voided as uric acid not amino acid. Similarly, in the TME and TAL bioassays excesses are stored as fat or lost as heat and do not affect the results. The assays usually involve precision- feeding a fasted bird with a known quantity of the test feedingstuff and collecting the resulting excreta. Each feedingstuff is fed at two, or more, levels of input to establish the relationship between nutrient input and output. For simplicity and convenience one input level is usually zero. Voluntary feeding has been used successfully as a substitute for precision feeding but it is subject to errors associated with feed spillage and consequent inaccurate estimates of intake. Furthermore, since birds eat at different rates the estimates of metabolic and endogenous losses may be subject to greater error. Fasted birds tend to catabolize more body protein than fed birds thus increasing metabolic energy excretion and introducing a bias into the TME assay. Similarly, the loss of body protein as energy containing excretory 10 - products is affected by the amount and composition of the test feedingstuf f , a secondary cause of bias. The problem is controlled by correcting the excreta energy outputs of both fed and fasted birds to zero nitrogen balance. The method for making the correction is explained later. A schematic representation of the partition of ingested feed energy by the bird is presented as Figure 2. The terminology and abbreviations are similar to those published by the National Research Council of the United States (256). The figure differs in certain respects from that published in the second edition of this Bulletin. The feces are assumed to comprise three energy containing fractions: the fecal energy of feed origin (F.E); the endogenous fecal energy (F E) ; and the metabolic fecal energy (F E) . The F E is contained in microflora and microbial debris and, for most practical e purposes, may be considered part of the F.E. The urine also comprises three energy containing fractions: the urinary energy of feed origin (U.E); the metabolic urinary energy (U E) ; and the endogenous urinary energy ID (U E) . The U E is contained in products of tissue catabolism, it being e m assumed that anything absorbed across the gut wall immediately becomes part of a tissue. The U E is contained in the excretory products resulting from e protein turnover. The partition shown in Figure 2 differs from that of the National Research Council (256) inasmuch as the F E and U E are included as part me of the net energy for maintenance. Further, the U E is placed under the m net energy for production because it results from tissue catabolism which may occur at the same time as tissue synthesis as well as in the absence thereof. The figure helps to explain the differences between AME and TME and shows that feces and urine contain energy components which are derived from the body rather than the feed. - 11 - Ingested Energy (IE) Apparent Digestible Energy (ADE) Fecal Energy (FE) Fecal Energy of Feed (F±E) Gaseous Energy (GE) Apparent Metabolizable Energy (AME) Urinary Energy (UE) Endogenous Metabolic Fecal Energy (FeE) Fecal Energy (V) L Endogenous Metabolic Urinary Urinary Energy (UeE) Urinary Energy (UmE) Energy of Feed (U±E) True Metabolizable Energy (TME) Net Energy (NE) Net Energy for Maintenance (NEm) Heat Increment (H^E) — Heat of Fermentation (HfE) — Heat of Digestion and Absorption (H^E) — Heat of Product Formation (HrE) — Heat of Waste Formation and Excretion (HWE) Net Energy for Production (NEr) — Basal Metabolism (HeE) — Heat of Activity (HjE) Heat of Thermal Regulation (HCE) — Metabolic Fecal Energy (FmE) — Endogenous Urinary Energy (UeE) — Tissue Growth — Fat Accretion — Carbohydrate Storage — Eggs — Semen — Metabolic Urinary Energy (UmE) Figure 2. The partition of ingested feed energy in the bird (the forerunner of this scheme is described in reference 336). 12 BIOASSAY PROCEDURES General The assays of the TME system have several procedures in common and may be performed simultaneously providing that there are sufficient excreta avail- able for the physical and chemical analyses. Birds are fasted, to empty their alimentary canals of feed residues, and then precision-fed a known quantity of the material to be assayed. Each bird is placed in a wire cage, where water is available ad libitum. The time is recorded and the excreta are col- lected quantitatively for a predetermined period of time. One bird in each replication remains fasted and serves as a negative control for the estimation of metabolic plus endogenous losses. The excreta, together with samples of the test materials, are assayed for gross energy, amino acids, lipid or mineral elements as appropriate. The basic calculation is as follows: TX = IX - (FX + UX) + (F X + U X + U X) m m e where: TX is the true available nutrient X; IX is the amount of X placed in the fed bird; (FX + UX) is the amount of X excreted by the fed bird; and (F X + U X + U X) is the amount of X excreted by the fasted bird, m m e The preferred bird is a dubbed adult cockerel of an egg-laying strain, which has never had access to grit. Other birds may be used but chicks have limited feed capacity, while fasted laying hens often produce shell-less eggs which tend to break and contaminate the excreta. Laying birds may be useful in assays for TAM where a high mineral requirement is desirable (338), but this is an atypical situation. Grit is avoided because it may be retained in the gizzard and voided on an irregular basis. Grit in excreta damages grinding equipment and introduces major errors into short-term mineral balances . All birds used in an assay must have been maintained on the same diet. The composition of the diet is not of critical importance providing that it allows the birds to satisfy their nutrient requirements. Many laboratories 13 - feed a laying hen diet, containing 15% of protein, during the maintenance period between assays. The preliminary fast is usually for 24 hours but a longer period may be required if the maintenance diet contains substantial quantities of indigestible materials. Where uncertainty exists it is advisable to measure the gut- clearance time of the maintenance diet before embarking on a series of assays. As the input of test material increases, the effects of experimental errors are reduced but the possibility of regurgitation increases; 30 to 40 g of air-dry feed is usually satisfactory. Large inputs, particularly of bulky feedingstuff s can lead to crop impaction. Impacted birds have extended feed residue retention times and consequently may yield misleading data. An exception to the foregoing is the TAM bioassay in which the input of the test mineral should be no greater than the bird's requirement because excess minerals are voided as minerals in the excreta. The use of fasted birds to measure metabolic plus endogenous losses is simple and convenient. The data obtained from a single group of such birds can be used to correct the excreta outputs of several other groups, in the same assay, fed a variety of feedingstuff s. The optimal number of fasted birds in an assay depends on several factors as discussed by Wolynetz and Sibbald (526). However, the use of fasted birds is not essential. The objective is to measure the relationship between the nutrient excreted and the nutrient input and this can be achieved by using two or more levels of input, none of which need be zero. Initially, it was recommended that the test material be pelleted but this is not necessary if the stem of the feeding funnel has an internal diameter of about 1.0 cm; however, care must be taken to avoid loss of feed by adherence to the funnel. Very dusty or hygroscopic materials are best fed in conjunction with a carrier, such as 90% ground maize plus 10% of vegetable oil; this makes it necessary to assay the carrier. The use of precision feeding is not essential although it is preferred. Assay data have been obtained successfully from birds allowed to consume feed voluntarily. - 14 - Test materials are weighed prior to an assay and held in containers until used. Clear polypropylene containers (130 ml) with close fitting lids such as are used for urine samples are satisfactory. Sub-samples of the test materials must be weighed for dry matter determination at the same time as the containers are being prepared. The timing is important because it avoids errors associated with water uptake or loss by the test materials. All analytical data describing the test materials should be expressed on the basis of dry matter. In this laboratory, birds are housed and maintained in individual wire cages on the lower level of a two-tier system. Cage design and construction are described later (page 22). Water is provided by a nipple system and feed is available in a trough running the full length of each block of cages. At the start of an assay, fasting is initiated by removal of the feed trough. Under other management systems, such as when water is provided in troughs, it is important that feed in the water system or adhering to the cages be removed. Fasted birds are taken from the lower tier of cages, given the appropriate treatment, and then housed in alternate cages of the upper tier. The technique of precision feeding is described later (page 17). The upper cages are used only during excreta collection periods and are scrupulously cleaned prior to each assay. Excreta collection trays are placed under each bird. The trays, preferably made of smooth plastic, are larger than the bottom of each cage thus reducing the chance of excreta loss. An alternative excreta collection technique is described later (page 28). Handling of birds causes losses of scale and feathers making quantitative excreta collection difficult. Blowing scale off the excreta collection trays an hour after housing reduces the problem. Excreta are collected at about 24 and again at exactly 48 hours after housing. A single 48 hour collection may be satisfactory but the double collection is favoured, when trays are used, because it reduces excreta deterioration and contamination. When the assays were first described a collection period of 24 hours was suggested but subsequent work showed this to be insufficient for clearance of the residues - 15 - of some test materials. Excreta adhering to the cage must be collected and excreta must be washed from any feathers trapped on the excreta collection tray. The trays must be checked for regurgitated feed which, if present, invalidates the use of the bird in the assay. The two excreta samples from each bird are frozen, dried, equilibrated with atmospheric moisture, weighed, pooled, ground, mixed and assayed. Freeze -drying is preferred because it leaves the excreta in a sponge like, easy- to-grind form; however, oven drying has been shown to be satisfactory for TME (97) and TAAA (370) bioassays. In some laboratories the excreta from several birds are pooled to reduce the analytical work. The procedure should not alter the estimated TME, TAAA, TAL or TAM values but it does restrict the ability to assess variation and to make comparisons between samples (343). Recent work has demonstrated the advisability of correcting TME values to zero nitrogen balance (TME ). As a first step in the calculations the n excreta energy output (FE + UE) is corrected to zero nitrogen balance (FE + UE ) , as follows: n n (FE + UE ) = (FE + UE) t k(lN - FN - UN) n n where: k is a constant which estimates the gross energy content of the excretory products resulting from the catabolism of a unit weight of tissue nitrogen; IN is the nitrogen input as test material; and FN and UN are the fecal and urinary nitrogen outputs. For fasted birds, IN is zero. In most assays the term k(IN - FN - UN) is negative; consequently, (FE + UE ) is usually smaller than (FE + UE) . n n The nitrogen -corrected energy excretion of a fasted bird is better described as(FE +UE +UE). TME values are calculated as follows: m n m n en n TME = IE - (FE + UE ) + (F E + U E 4 U E ) n n n mnmnen where: IE is the amount of energy, as test material, placed in the bird. There is no evidence that similar corrections are required in any of the other bioassays of the TME system. - 16 Precautions The following is a list of precautions to be observed if high quality assay data are to be obtained. The list concludes with the most common causes of abnormally large and small values. 1. Assay birds must be in good health. 2. All assay birds must be fed the same maintenance diet between assays. 3. Assay birds should be grit-free. 4. Test materials must be assayed for dry matter at the time that they are weighed into containers in preparation for feeding to the birds. 5. Dusty and hygroscopic test materials should be fed in conjunction with a carrier; the carrier must also be assayed. 6. Assay birds must be fasted for sufficient time to allow all feed residues to be voided. 7. Feed removal for fasting must be total. Feed adhering to the cage and usually ignored by the bird will be eaten if no other is available. 8. Birds must have continuous access to fresh water. 9. Excreta trays must be checked for regurgitated feed which, when found, eliminates the bird from the assay. 10. The excreta collection period must be exactly the same for all birds in an assay. 11. When using adult cockerels, and feed inputs of 30 to 40 g, an excreta collection period of 48 hours should be sufficient. For other birds and inputs a preliminary experiment may be necessary to establish the length of the collection period. 12. Excreta collection must be quantitative. Feathers must not be included and scale is to be avoided as much as possible. 13. Dried excreta should be equilibrated with atmospheric moisture or be held in such a manner that its moisture content remains constant between weighing and sub-sampling for analysis. Abnormally large values may result from: a. incomplete clearance of the residues of the test material, check for crop impaction; b. incomplete excreta collection, excreta may have been voided beyond the collection tray; 17 - c. unobserved regurgitation beyond the collection tray causing excreta output to be too small; d. errors in weighing, preparing or administering the test material; and e. analytical errors. Abnormally small values are obtained when: a. the preliminary fast is inadequate and residues of the maintenance diet are assumed to be derived from the test material; b. the bird has access to feed during the fast; c. regurgitated feed is mixed with the excreta; d. scale or feathers are included with the excreta; and e. preparatory and analytical errors. The assays are simple and relatively fast. However, like all assays they require care and attention to detail if high quality data are to be obtained. Precision Feeding The purpose of precision feeding is to ensure that a known quantity of a feedingstuff enters the alimentary canal of a bird at a known time. The procedure avoids the need to recover waste feed, prevents feed selection and eliminates variation of intake among birds. All of these problems are encountered when birds ingest feed voluntarily. Precision feeding involves insertion of a tube from the beak, via the esophagus, into the crop, causing feed to move through the tube, and removal of the tube. The initial method developed at the Animal Research Centre used a simple glass tube into which pelleted feed was placed and then pushed into the crop with a glass rod. From this was developed a glass funnel and metal plunger. With experience came the ability to insert tubes with larger diameters and an effective device was prepared by taping a glass powder funnel to a piece of 1.2 cm diameter copper water pipe. A metal rod was used as a plunger. Today, a stainless steel funnel is used with a stem 40 cm long, an external diameter of 1.2 cm and an internal diameter of 1.1 cm. The plunger consists of a 1.0 cm diameter aluminum rod to which a 3.0 cm diameter spherical knob is attached. A plastic sleeve is riveted to the rod to 18 - prevent the plunger from projecting more than 0.5 cm beyond the end of the funnel. Ease of operation demands that the funnel be light in weight and well balanced. Heavy funnels, particularly those with uneven weight distribution, are difficult to control when in the bird. Plastic funnels have the advantage of light weight but many possess electrostatic properties which make quantitative feed delivery impossible. Some of the feeding devices which have been used in this laboratory are shown in Figures 3 and 4. Successful precision feeding depends upon control of the bird. A simple, rapid technique used at the Animal Research Centre is as follows. The operator sits on a stool, or chair without sides, and crosses his left leg over his right. A bird is taken with both hands and placed with the keel (breast bone) pushed into the groove formed by the left thigh and abdomen of the operator. The body of the bird is at 45 degrees from the vertical. The legs of the bird project to the left and are unable to gain leverage on anything. The bird is held firmly in place with the elbow of the left arm, and the beak is grasped from above the head with the left thumb and fore- finger. The neck of the bird is slightly extended, the beak is opened and the stem of the funnel is inserted. The funnel should move down the esophagus and into the crop with a minimum of effort. If a blockage is encountered, wait a few seconds for the bird to swallow or relax. If this is unsuccessful, remove the funnel and try again. Feed is poured into the funnel from a container held in the right hand and pushed into the crop by a second operator. If the funnel tends to move out of the bird as feed is pushed down, the end of the funnel is in the esophagus not the crop. Deposition of feed in the esophagus can lead to regurgitation. The funnel is withdrawn using the right hand. The left hand is moved to the neck of the bird and gentle pressure is applied to the esophagus. This, coupled with rotary withdrawal of the funnel, removes feed particles which may have adhered to the outside of the stem of the funnel. The complete operation usually takes less than one minute per bird. The precision feeding technique is illustrated in Figures 5 to 12. A six-minute, 16 mm, colour, sound film entitled "Poultry Force Feeding - An Experimental Technique" is available by writing to: Communications Branch, Agriculture Canada, Ottawa, Ontario, Canada, K1A 0C7 . - 19 Figure 3. Some early feeding devices: (A) glass funnel and rod, (B) copper water pipe attached to a glass powder funnel and an aluminum plunger, (C) simple glass tube and rod. Figure 4. Stainless steel funnel (A) with aluminum plunger (B) Note plastic sleeve (C) on plunger which controls penetration (D) beyond the end of the funnel. 20 Figure 5. Bird is placed at a 45° angle with keel in groove formed by the operator's thigh and abdomen. Figure 6. The legs project and cannot gain leverage on anything The bird is held in place by the left elbow. wm sR^I f V • 1 J* Figure 7. The beak is opened from above using the thumb and forefinger of the left hand. Figure 8. The neck is slightly extended and the stem of the funnel inserted. 21 - Figure 9. The inserted funnel is Figure 10 held in place by the thumb funnel, and forefinger. Feed is poured into the Figure 11. Feed is pushed down the Figure 12. The funnel is removed funnel by a second operator. Note with a rotary action, pressure is that the first operator helps to applied to the esophagus with the hold the funnel in place. left hand to dislodge feed particles adhering to the stem of the funnel. - 22 - Cage Design and Construction The cages used to house the adult cockerels have the following dimensions: depth (front to back.) 40.6 cm, height 50.8 cm and width 30.5 cm. Figures 13 to 21 describe the cages, their assembly and use; all dimensions are in centimetres. The back, top and floor of a row of 10 cages is fabricated from a single piece of wire mesh approximately 1.3 m wide x 3.05 m long. The wire is 2.03 mm diameter (14 gauge). The mesh is a 5.1 x 2.5 cm grid welded at each corner. The internal partitions and ends are made of the same material. The cage fronts, also 3.05 m in length, are made from 4.11 mm diameter (8 gauge) wire. The door opening of 23 x 28 cm provides adequate access to remove the birds. The cages are set up in a double-deck arrangement and the droppings board is placed horizontally to hold the excreta collection trays. Water is available from a nipple system on the top of the cages. 23 Figure 13. End view of the single piece of wire mesh which is folded to create the body (top, back and bottom) of a row of 10 cages. 40 2.5 5.1 5 \ i 08 1 Figure 14. A typical cage partition or end. Partitions are located at 30.5 cm intervals and are welded in place. - 24 - -305 30.5 .1 - -— 13.81- 23- 3.8- — 23 DOOR OPENING 28 ® 23j 1 76 i 2.5 50.8 6.4 -4 3.8 THIS HORIZONTAL $ .WIRE SHAPED OUT 0.5 TO ACT AS A DOOR GUIDE Figure 15. A. The front for a row of cages. B. Side view of cage front. C-N Figure 16. ® A. The front with cage doors installed B. Side view of cage door. C. Detail of cage door. ® ' SHAPED OUT 0.5 BOTTOM BAR OF CAGE DOOR HOOKS AROUND DOOR GUIDE c - 25 Figure 17. The relationship between the cage body with partitions and the front. For simplicity, only 2 doors are shown on the front. Figure 18. An assembled cage unit with feed trough. 26 - X I , .J \ \ ____ \ .. % p (Ti \oJ i \ \ \ \ SUPPORT BAR 3 isi ; _t -t i ' ■^ —~~ — ,< — > ::::::::::::::::: / ====^= (r\ (Jf I ) ::::::::::::::::: / / SUPPORT FOR FEED TROUGH Figure 19 Side view of a 4-row double deck battery of cages showing, (A) position of droppings board to hold excreta collection trays and (B) normal position of droppings board. - 27 Cages being used to maintain birds between assays. Figure 21. Birds housed in cages of upper tier being used in an assay. 28 - Excreta Collection Initially, excreta were collected on smooth plastic trays which were larger than the bottoms of the cages. The procedure continues to be used in many laboratories but has the disadvantage that scale and feathers may be trapped in the excreta. There is also the possibility of excreta falling beyond the edges of the trays. A procedure whereby human colostomy bags are attached to birds to collect excreta was described in the previous edition of this Bulletin. After working with the procedure in a number of assays it was abandoned because: a) there was a high incidence of adhesive failure and samples were lost due to bags falling off the birds; and b) removal of those bags which remained attached caused feathers to be pulled out so that the area around the cloaca was denuded. A harness designed to hold an excreta collection bag has been developed in Portugal (444). Modifications have been made to the design and the harness is now used routinely at the Animal Research Centre. The major modification is the addition of two short tapes which are tied around the base of the tail to prevent vertical slippage. When using the harness four holes are cut in the collection bag and the tapes are threaded through them. The neck of the bag is then folded back. With this procedure the use of adhesive tape to hold the bag to the harness is avoided. When the harness is on the bird, the ends of the two long tapes are tied together as a precaution against lateral slippage. The harness, its method of attachment, and its use are illustrated in Figures 22 to 35. At the Animal Research Centre, the collection bags are labelled, holes for the tapes are cut, the weight of each bag is recorded, and the bags are attached to the harnesses. The harnesses, with bags, are attached to the birds immediately after precision feeding. At the conclusion of an excreta collection, the harnesses are removed from the birds and the bags plus excreta are separated from the harnesses. The excreta are frozen, freeze- dried, and equilibrated with atmospheric moisture while in the bags. The procedure is efficient and provides excreta free of scale and feathers. 29 - Care is required when attaching a harness to a bird. If the tension of the tapes is too great the bird has difficulty maintaining its balance and bruising appears in the wings. Even when the minimum required tension is used there is a temporary disturbance of balance for one or two minutes. Birds do peck, at the bags and can cause additional damage with their spurs. Debeaking and spur removal reduces the problem and has the additional benefit of making the birds easier to handle. Detection of leaking bags, and of regurgitation, is made easier if excreta collection trays are placed below harnessed birds. Clipping the feathers around the cloaca is recommended to prevent excreta adherance. - 30 Figure 22. The excreta collection harness comprises a plastic ring (OD 7.5 cm, ID 5.75 cm) (A), 2 long cotton tapes (40 cm) (B), 2 short cotton tapes (20 cm) (C) and 2 elastic tapes (10 cm) (D) each attached to two small plastic rings (OD 1.9 cm) (E) which serve as buckles. Each D is sewn to the corresponding C at F. All the tapes are 1.25 cm wide and the ends of the cotton tapes are varnished to prevent fraying. Figure 23. The tapes of the harness are threaded through holes in a labelled and weighed plastic bag which measures 15.2 x 25.4 cm. x .10 mm thick (see also Figure 34). Figure 24. Harness and bag about to be attached to the bird. The first step is to attach the short tapes around the base of the tail. Note that the neck of the bag has been everted over the large ring. - 31 - Figure 25. Tying the short tapes Figure 26. between the and is then shoulder. A long tape is placed thigh and the abdomen brought around the Figure 27. Each long tape is buckled to the elastic tape on the opposite side of the back using the pair of small rings. When in place these tapes form a cross on top of the back. Figure 28. The ends of the long tapes are tied together to reduce the chance of lateral slippage. - 32 Figure 29. The location of the Figure 30. The location of the plastic ring when the harness is collection bag when the harness in place. The collection bag has is attached, been removed. Note that the feathers around the cloaca have been clipped. J •5k ■p Figure 31. Bird immediately after attaching bag and harness. - 33 - Figure 32. Bird with plastic bag containing excreta. Figure 33. Rear view of bird showing collected excreta. Figure 34. Bag and harness removed from bird. Figure 35. Bag with excreta ready for further processing 34 FEEDINGSTUFF COMPOSITION Changes in feedingstuf fs over time and the introduction of new analytical procedures combine to create a continuing demand for data describing feedingstuff composition. The second edition of this Bulletin contained a simple table listing the proximate composition, gross energy, TME and TME n values of a variety of feed ingredients. The information has now been expanded in two directions. The original table has been revised and now includes data describing more than 200 additional samples; for many of the new samples calcium, total phosphorus and neutral detergent fibre values are presented. Three new tables present total and true available amino acid values for many of the new samples. Data are expressed as g/kg or MJ/kg of dry matter except in Table 4 where percentages are used. Samples of feedingstuf fs were obtained from several sources including commercial feed manufacturers, the Canadian Wheat Board, plant breeders and the Animal Research Centre Research Farm. On receipt, each sample was given a code number and all available information, descriptive of the sample, was entered in a catalogue. The samples were stored in sealed containers in a freezer until required for analysis. At the start of a bioassay the required number of samples was withdrawn from storage, in the order of their receipt, and allowed to warm to room temperature. Coarse samples, such as whole grains, were ground. Each sample was thoroughly mixed and sub- samples were taken for the bioassay and for dry-matter measurement. If a sample was finely divided, additional sub-samples were taken for physical and chemical analysis. Alternatively, less fine material was reground prior to sub- sampling for the physical and chemical analyses; when this step was introduced new dry-matter measurements were made to minimize errors associated with variation in water content. The feedingstuf fs were assayed for dry matter, ether extract, nitrogen, crude fibre, ash and calcium using the procedures of the A.O.A.C. (Official Methods of Analysis, 1980). Phosphorus was measured by Hambleton's method - 35 (Advances in Automated Analysis, 7th Technicon Int. Conf., 1976) and neutral detergent fibre by the procedure of Van Soest (J.A.O.A.C. 50:50-55, 1967). Gross energy was measured with a Parr adiabatic oxygen bomb calorimeter fitted with a 'master control' unit. Amino acids were measured with a Beckman 121M automatic analyzer following hydrolysis with 6N HC1 (J. Agr. Food Chem. 13:266-288, 1965). A performic acid digestion (J. Biol. Chem. 238:235-237, 1963) was used in the analysis for sulphur amino acids. The TME and TME values were measured as described herein. The n preliminary fast was 24 hours, the feed input was usually 30 g of air-dry material per bird, and the excreta collection period was 48 hours. Each TME and TME value is the mean of 6 to 8 replicated observations. The TAAA n assays were made using the excreta samples collected during the TME assays; however, excreta from birds given a particular feedingstuff were pooled, in proportion to the amounts voided by individual birds, to yield two composite samples each containing excreta from three or four birds. Amino acid profiles were obtained for each pooled sample and used to compute two sets of TAAA data from which the means of Tables 3 and 4 were calculated. Amino acid analyses of feedingstuff s were made in duplicate. In Table 4 there are 18 values in excess of 100%. Most describe the bioavailabilities of arginine and lysine in ground yellow corn. Such values are erroneous and probably arose because the estimates of metabolic and endogenous amino acid losses, measured with fasted birds, differed slightly from those of the fed birds. Examination of the relevant duplicate values obtained with fasted birds showed them to be more variable and to have larger means than average. In other experiments (57, 95) the bioavailabilities of corn amino acids have approached 100% so it is reasonable that even small errors could result in substantial over-estimates. Additional factors possibly contributing to error are the low amino acid concentrations in corn and the relatively high lysine concentrations in the excreta of fasted birds. Each feedingstuff is described in Table 1. Only abbreviated descriptions appear in Tables 2, 3 and 4 but reference to Table 1 descriptions is possible - 36 - by use of the code numbers which appear in the left hand column of each table. The code numbers also serve to allow readers to identify information added since the second edition (621-862). Those who have included the earlier table in their data banks will wish to avoid double entry of information and the consequent bias. The nitrogen data are recorded as g/kg of dry matter. No attempt was made to estimate protein concentrations because the validity of some conversion factors is debatable. Recently, the assay for crude fibre was replaced by that for neutral detergent fibre; the latter assay does not apply to animal products and their fibre values are no longer estimated. Compositional means for feedingstuf f s are not presented because they may be misleading if an anomalous sample in included. The term 'whole ground' used to describe some grains means that whole, intact kernels were ground and assayed; there was no intentional classification or separation of the ground product. The term 'hull-less' denotes a genotype which produces a grain with little or no hull and should not be confused with 'dehulled' which implies physical removal of the hull. Oat groats, for example, are dehulled oats. There are many sources of additional data which could be of interest to those constructing data banks. Many references will be found in the bibliography. The data recorded in Tables 1, 2, 3 and 4 were obtained within the facilities of Agriculture Canada subsequent to publication of a table appearing in reference 32. - 37 - n >. Uh CO 60 0 to e >i l, •o 60 A! 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Abstracts are identified by asterisks. Some references may have been overlooked and readers are asked to bring them to the attention of the author for inclusion in subsequent updates or revisions. 191/ 1 Armsby, H.P. , 1917. The Nutrition of Farm Animals. The MacMillan Co., New York, U.S.A. 1924 2 Mitchell, H.H., 1924. A method for determining the biological value of protein. J. Biol. Chem. 58:873-903. 1926 3 Kellner, 0., 1926. The Scientific Feeding of Animals (Translated by W. Goodwin). Duckworth, London, U.K. 1966 4 Harris, L.E., 1966. Biological Energy Interrelationships and Glossary of Energy Terms. N. A.S.-N.R.C. Publ . 1411, Washington, U.S.A. 1970 5 Guillaumo, J., and J.D. Summers, 1970. Maintenance energy requirement of the rooster and influence of plane of nutrition on metabolizable energy. Can. J. Anim. Sci. 50:363-369. 1975 6 Sibbald, I.E., 1975. The effect of level of feed intake on metabolizable energy values measured with adult roosters. 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Comparison of methods for the bioassay of the metabolizable energy of wheat-based poultry diets. Proc . Poult. Husb. Res. Found. Symp., Sydney, Australia. 6 pp. 303 Muztar, A.J., and S.J. Slinger, 1982. The true metabolizable energy and amino acid content of Candle, Altex and Regent Canola meals. Can. J. Anim. Sci. 62:521-525. 304 Ngoupayou, J.D.N. , P.M. Maiorino and B.L. Reid, 1982. Jojoba meal in poultry diets. Poultry Sci. 61:1692-1696. 305 Okumura, J., Y. Isshiki and Y. Nakahira, 1982. Influence of dietary cellulose and indigestible dry matter on metabolic and endogenous nitrogen excretion in chickens. Jpn . Poult. Sci. 19:300-303. 306 Parsons, CM., L.M. Potter and B.A. Bliss, 1982. True metabolizable energy corrected to nitrogen equilibrium. Poultry Sci. 61:2241-2246. 307 *Parsons, CM., L.M. Potter and B.A. Bliss, 1982. True metabolizable energy corrected to nitrogen equilibrium. Poultry Sci. 61:1523. 308 Parsons, CM., L.M. Potter and R.D. Brown, Jr., 1982. 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