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Influence of a Fat-Mobilizing Substance from Geese Urine on Geese and Chickens
629
RESEARCH NOTES
The results of this study indicated that rapeseed meal has greater zinc binding capacity than soybean and that the zinc binding capacity of rapeseed meal appears to be related to its content of bound tannins or to some, as yet, unidentified factor(s) in the meal rather than to its content of methanol extractable tannins.
REFERENCES Clandinin, D. R., and J. Heard, 1968. Tannins in prepress-solvent and solvent-processed rapeseed meal. Poultry Sci. 47: 688-689. Jurd,L.,andT. A. Geissman, 1956. Absorption spectra of metal complexes of flavanoid compounds. J. Org. Chem. 27: 1395-1401. Steel, R. G. D., and J. H. Torrie, 1960. Statistical Principles in Experimental Design. McGraw-Hill Book Company, Inc., New York. Yapar, Z., and D. R. Clandinin, 1972. Effect of tannins in rapeseed meal on its nutritional value for chicks. Poultry Sci. 51: 222-228.
INFLUENCE OF A FAT-MOBILIZING SUBSTANCE FROM GEESE URINE ON GEESE AND CHICKENS I . N l R 1 AND ZAFRIRA NlTSAN 2
Department of Animal Science, Faculty of Agriculture, Hebrew University of Jerusalem, and2 Division of Poultry Science, Agricultural Research Organization, The Volcani Center, Rehovot, Israel (Received for publication October 1, 1974)
ABSTRACT A fat-mobilizing substance (FMS) was extracted from the urine of fasting, ad-libitum-fed or over-fed colostomized geese. Injection of these materials into geese and cockerels caused increases in plasma-free fatty acids (FFA). A marked anorexigenic effect was obtained in cockerels only, by the FMS obtained from ad-libitum-fed geese. The daily amount of FMS excreted in the urine was proportional to the amount of feed consumed. POULTRY SCIENCE 54: 629-632, 1975
MATERIALS AND METHODS
P
REPARATION of FMS. Two adult geese from a local breed already described (Nir and Perek, 1971) were prepared with colostomies by the method of Ariyoshi and Morimoto (1956). They were kept in metabolic cages and the urine was collected
in containers kept on crushed ice. FMS was extracted from pooled samples as described by Nir et al. (1969). The diet used was a 16% protein commercial mash. When force fed, a mash composed of corn and 10% soybean meal was used (Nir et al., 1973). Force feeding was performed with a funnel
Downloaded from http://ps.oxfordjournals.org/ at Univ of Iowa-Law Library on June 7, 2015
soybean meals (83.6% vs. 50.3%) and that different samples of rapeseed meal or of soybean meal may differ significantly in their zinc binding capacity. Zinc uptakes of the rapeseed meals which had been subjected to tannin extraction, presented in Table 3, indicate that removal of the soluble polyphenols significantly decreased the zinc binding properties of the meals. It would appear, however, that the tannins (Table 2) that remained in the meals after methanol extraction or some, as yet, undetermined factors in the meals were more important in the retention of zinc than the tannins which were extracted by the procedure followed. No zinc was found in the tannins precipitated by lead acetate. Any zinc that complexed with these polyphenols compounds was probably replaced by lead ions during the precipitation process and washed out of the precipitates.
30 ± 3 261 ± 6
31 ± 5 262 ± 2
23 ± 268 ±
29 ± 2 270 ± 2
137 ± 6 37 ± 4
146 + 4 37 ± 3 24 ± 2 261 ± 7
Recipients Cockerels (6) 136 + 4 128 ± 7 145 ± 34 + 3 45* ± 3 46 +
(6)
82 ± 1 219 ± 1
115 ± 15 227 ± 1 1
131 ± 12 264* ± 10
156 ± 42 210 ± 9 115 ± 9 206 ± 3
104* ± 13 47 ± 4
181 ± l l n 40 ± 5
(9)
PostPreinjection injectio Recipients Geest (9) 97* ± 11 197 ± 9 210 ± 4 66* ± 8 32 ± 2 38 ± 7
Starved geese Preinjection
NOTE: Figures in parentheses represent number of animals assayed. *A11 groups designated with an asterisk are statistically different from the pre-injection value at the P < 0.05 level. *Pre injection, average of 4 days preceding injection; Post injection, 24 intake following injection. tf Mean ± SE of the mean.
Food intake* (g./24h.) Plasma FFA (meq./100 ml.) Plasma triglycerides (mg./lOO ml.) Plasma glucose (mg.%)
Food intake* (g./24h.) Plasma FFA (meq./KX) ml.) Plasma triglycerides (mg./lOOml.) Plasma glucose (mg.%)
Measurement of
Preinjection
Postinjection
0.5% N a 2 C 0 3
A f
F
TABLE 1.—Effect of a single intravenous injection of 0.5% Na2C03 (carrier) or of 0.5 mg./kg.-FMS ex over-fed geese on food intake, plasma FFA, triglycerides and blood plasma glucose of geese and cockerels injection)
ed from http://ps.oxfordjournals.org/ at Univ of Iowa-Law Library on June 7, 2015
631
RESEARCH NOTES
TABLE 2.- -Feed intake, urine and FMS output during starving, ad libitum-/ee![]()
(average of 2 geese)
Starving (4)* Ad libitum-feeding (5) Over-feeding (4)
FMS excreted in urine
Feed intake (g-/day)
Urine output (ml. /day)
(mg./l.)
(mg./goose/day)
151 560
260 360 1510
48.8 42.7 151.0
12.7 15.4 228.0
*Numbers in parentheses indicate days of urine collection. used in cramming geese for the production of goose fatty liver.
Chemical Methods. Blood plasma glucose was determined by the glucose oxydase procedure with the Biochemica Test Combination (C. F. Bochringer und Soehne GmbH, Mannheim, W. Germany). Blood plasma FFA and triglyceride were determined as described earlier (Nir et al., 1969). RESULTS AND DISCUSSION As with the FMS obtained from chicken urine (Nir et al., 1969), the FMS, obtained from the urine of starved, ad-libitum-fed or over-fed geese, had lipid-mobilizing properties. When injected into cockerels or geese the blood plasma FFA concentration was consistently increased. The effect of the different FMS preparations on plasma triglycerides was not consistent (Table 1). When injected into geese the anorexigenic property of the FMS could not be demonstrated. In the geese, injection and handling were stresses which reduced feed intake and increased plasma glucose (Table 1). It should be pointed out that when FMS prepared from fasted cockerels was injected, an anorexigenic effect of the material could be demonstrated in the geese, in spite of their excessive sensitivity to handling during blood drawing (Nir and Levy, 1972). In the cited work the feed consumption of the control birds was
REFERENCES Ariyoshi, S., and H. Morimoto, 1956. Studies on the nitrogen metabolism in the fowl. I. Separation of
Downloaded from http://ps.oxfordjournals.org/ at Univ of Iowa-Law Library on June 7, 2015
Assay of FMS activity. The care and treatment of the recipient birds was already described (Nir et al., 1969; Nir and Levy, 1972).
reduced by about 50% and in the treated it was reduced by 75%. According to their effect on the plasma FFA concentration, the different preparations had a similar fat-mobilizing activity in the geese. In cockerels a difference was observed in the activity of the different materials. The FMS obtained from the ad-libitum-fed geese was the most active on fat mobilization (plasma FFA increase) and food intake reduction. The FMS obtained from the starved or overfed geese caused an increase in blood plasma FFA concentration but had no anorexigenic effect. This result is surprising since, as stated by Weil and Stetten (1947) and confirmed in mammals (Chalmers, 1965; Stevenson et al., 1964) and chickens (Nir et al., 1969), the material extracted from fasted animals should be the most active. The marked increase in the amount of FMS excreted during over-feeding (Table 2) agrees well with the observation of Mukaida and Lichton (1971), that FMS 1A excretion by rats was proportional to the amount of food consumed, and of Beaton et al. (1966), that higher FMS 1A was excreted by fed rats compared to fasted rats. It is likely that the geese FMS preparations contain both of the separate anorexigenic and lipid mobilizing substances identified by Beaton et al. (1966). The amount of food intake probably affects the excretion and potency of each and could explain some of the results observed.
632
RESEARCH NOTES
Nir, I., Z. Nitsan and A. Vax, 1973. The influence of force feeding and of protein supplementation to the diet on the metabolizable energy of diets, digestibility of nutrients, nitrogen retention and digestive enzymes output. Ann. Biol. Anim. Bioch. Biophys. 13: 465-479. Nir, I., and M. Perek, 1971. The effect of various protein levels in feed of goslings during the preparatory period on fatty liver production and blood plasma components. Ann. Biol. Anim. Bioch. Biophys. 11: 645-656. Stevenson, J. A. F., B. M. Box and A. J. Szlavko, 1964. A fat-mobilizing and anorectic substance in the urine of fasting rats. Proc. Soc. Exptl. Biol. Med. 115:424-429. Weil, R., and De W. Stetten, Jr., 1947. Urinary excretion of a fat-mobilizing agent. J. Biol. Chem. 168: 129-132.
A THEORETICAL METHOD FOR T H E DETERMINATION O F AMINO ACID A V A I L A B I L I T Y U T I L I Z I N G 14C L A B E L E D P R O T E I N S O U R C E S ' S. V. A M A T O , 2 R. M . FORBES AND H . M . SCOTT
Department of Animal Science, University of Illinois, Urbana, Illinois 61801 (Received for publication October 4, 1974)
ABSTRACT A theoretical method is described whereby the availabilities of amino acids from test proteins are determined relative to the availabilities of amino acids from egg albumen. The method requires that both the test protein and the egg albumen be ,4C labeled. The relative ' availabilities of the amino acids are determined from the change in specific activities of the dietary amino acids and those recovered in the feces. POULTRY SCIENCE 54: 632-634, 1975
INTRODUCTION
A
LTHOUGH the determination of availability of amino acids from protein supplements has been under study for many years, no completely satisfactory method has yet been developed. The most commonly used methods as well as the shortcomings
1. Based on a thesis submitted to the Graduate College of the University of Illinois is partial fulfillment of the requirement for the Ph.D. degree. 2. Present address: Gold Kist Inc., Atlanta, Georgia 30301.
of each have been reviewed by De Muelenaere et al. (1967). In the present communication a theoretical method for the determination of availability of amino acids is described which avoids many of the problems discussed by De Muelenaere et al. (1967). This method is based on the procedure described by Amato (1960) which involves the feeding of a ,4C labeled protein reference standard and l4C labeled test proteins to different groups of animals. The availability of the amino acids of the test proteins is determined relative to that of the reference protein and is based on the
Downloaded from http://ps.oxfordjournals.org/ at Univ of Iowa-Law Library on June 7, 2015
urine for nutritional balance studies. Bull. Natl. Inst. Agr. Sci. Ser. G. 12, 37-44. Beaton, J. R., and A. J. Szlavko and J. A. F. Stevenson, 1966. Factors influencing the excretion of a fat-mobilizing substance in the urine of rats. Can. J. Physiol. Pharmacol. 44: 95-101. Chalmers, T. M., 1965. Lipid-mobilizing activity during fasting. In: Handbook of Physiology. Edited by A. E. Renold and G. F. Cahill, Jr. Waverly Press, Inc., Baltimore, Md. pp. 549-555. Mukaida, C. S., and I. J. Lichton, 1971. Some dietary influences on the excretion and biological activity of an anorexigenic substance in the urine of rats. J. Nutr. 101:767-774. Nir, I., M. K. Dimick and S. Lepkovsky, 1969. A fat-mobilizing substance in chicken urine. Can. J. Physiol. Pharmacol. 47: 435-443. Nir, I., and V. Levy, 1972. Influence of a fat-mobilizing substance from chicken urine on geese. Poultry Sci. 51: 1041-1042.
RESEARCH NOTES
The results of this study indicated that rapeseed meal has greater zinc binding capacity than soybean and that the zinc binding capacity of rapeseed meal appears to be related to its content of bound tannins or to some, as yet, unidentified factor(s) in the meal rather than to its content of methanol extractable tannins.
REFERENCES Clandinin, D. R., and J. Heard, 1968. Tannins in prepress-solvent and solvent-processed rapeseed meal. Poultry Sci. 47: 688-689. Jurd,L.,andT. A. Geissman, 1956. Absorption spectra of metal complexes of flavanoid compounds. J. Org. Chem. 27: 1395-1401. Steel, R. G. D., and J. H. Torrie, 1960. Statistical Principles in Experimental Design. McGraw-Hill Book Company, Inc., New York. Yapar, Z., and D. R. Clandinin, 1972. Effect of tannins in rapeseed meal on its nutritional value for chicks. Poultry Sci. 51: 222-228.
INFLUENCE OF A FAT-MOBILIZING SUBSTANCE FROM GEESE URINE ON GEESE AND CHICKENS I . N l R 1 AND ZAFRIRA NlTSAN 2
Department of Animal Science, Faculty of Agriculture, Hebrew University of Jerusalem, and2 Division of Poultry Science, Agricultural Research Organization, The Volcani Center, Rehovot, Israel (Received for publication October 1, 1974)
ABSTRACT A fat-mobilizing substance (FMS) was extracted from the urine of fasting, ad-libitum-fed or over-fed colostomized geese. Injection of these materials into geese and cockerels caused increases in plasma-free fatty acids (FFA). A marked anorexigenic effect was obtained in cockerels only, by the FMS obtained from ad-libitum-fed geese. The daily amount of FMS excreted in the urine was proportional to the amount of feed consumed. POULTRY SCIENCE 54: 629-632, 1975
MATERIALS AND METHODS
P
REPARATION of FMS. Two adult geese from a local breed already described (Nir and Perek, 1971) were prepared with colostomies by the method of Ariyoshi and Morimoto (1956). They were kept in metabolic cages and the urine was collected
in containers kept on crushed ice. FMS was extracted from pooled samples as described by Nir et al. (1969). The diet used was a 16% protein commercial mash. When force fed, a mash composed of corn and 10% soybean meal was used (Nir et al., 1973). Force feeding was performed with a funnel
Downloaded from http://ps.oxfordjournals.org/ at Univ of Iowa-Law Library on June 7, 2015
soybean meals (83.6% vs. 50.3%) and that different samples of rapeseed meal or of soybean meal may differ significantly in their zinc binding capacity. Zinc uptakes of the rapeseed meals which had been subjected to tannin extraction, presented in Table 3, indicate that removal of the soluble polyphenols significantly decreased the zinc binding properties of the meals. It would appear, however, that the tannins (Table 2) that remained in the meals after methanol extraction or some, as yet, undetermined factors in the meals were more important in the retention of zinc than the tannins which were extracted by the procedure followed. No zinc was found in the tannins precipitated by lead acetate. Any zinc that complexed with these polyphenols compounds was probably replaced by lead ions during the precipitation process and washed out of the precipitates.
30 ± 3 261 ± 6
31 ± 5 262 ± 2
23 ± 268 ±
29 ± 2 270 ± 2
137 ± 6 37 ± 4
146 + 4 37 ± 3 24 ± 2 261 ± 7
Recipients Cockerels (6) 136 + 4 128 ± 7 145 ± 34 + 3 45* ± 3 46 +
(6)
82 ± 1 219 ± 1
115 ± 15 227 ± 1 1
131 ± 12 264* ± 10
156 ± 42 210 ± 9 115 ± 9 206 ± 3
104* ± 13 47 ± 4
181 ± l l n 40 ± 5
(9)
PostPreinjection injectio Recipients Geest (9) 97* ± 11 197 ± 9 210 ± 4 66* ± 8 32 ± 2 38 ± 7
Starved geese Preinjection
NOTE: Figures in parentheses represent number of animals assayed. *A11 groups designated with an asterisk are statistically different from the pre-injection value at the P < 0.05 level. *Pre injection, average of 4 days preceding injection; Post injection, 24 intake following injection. tf Mean ± SE of the mean.
Food intake* (g./24h.) Plasma FFA (meq./100 ml.) Plasma triglycerides (mg./lOO ml.) Plasma glucose (mg.%)
Food intake* (g./24h.) Plasma FFA (meq./KX) ml.) Plasma triglycerides (mg./lOOml.) Plasma glucose (mg.%)
Measurement of
Preinjection
Postinjection
0.5% N a 2 C 0 3
A f
F
TABLE 1.—Effect of a single intravenous injection of 0.5% Na2C03 (carrier) or of 0.5 mg./kg.-FMS ex over-fed geese on food intake, plasma FFA, triglycerides and blood plasma glucose of geese and cockerels injection)
ed from http://ps.oxfordjournals.org/ at Univ of Iowa-Law Library on June 7, 2015
631
RESEARCH NOTES
TABLE 2.- -Feed intake, urine and FMS output during starving, ad libitum-/ee
(average of 2 geese)
Starving (4)* Ad libitum-feeding (5) Over-feeding (4)
FMS excreted in urine
Feed intake (g-/day)
Urine output (ml. /day)
(mg./l.)
(mg./goose/day)
151 560
260 360 1510
48.8 42.7 151.0
12.7 15.4 228.0
*Numbers in parentheses indicate days of urine collection. used in cramming geese for the production of goose fatty liver.
Chemical Methods. Blood plasma glucose was determined by the glucose oxydase procedure with the Biochemica Test Combination (C. F. Bochringer und Soehne GmbH, Mannheim, W. Germany). Blood plasma FFA and triglyceride were determined as described earlier (Nir et al., 1969). RESULTS AND DISCUSSION As with the FMS obtained from chicken urine (Nir et al., 1969), the FMS, obtained from the urine of starved, ad-libitum-fed or over-fed geese, had lipid-mobilizing properties. When injected into cockerels or geese the blood plasma FFA concentration was consistently increased. The effect of the different FMS preparations on plasma triglycerides was not consistent (Table 1). When injected into geese the anorexigenic property of the FMS could not be demonstrated. In the geese, injection and handling were stresses which reduced feed intake and increased plasma glucose (Table 1). It should be pointed out that when FMS prepared from fasted cockerels was injected, an anorexigenic effect of the material could be demonstrated in the geese, in spite of their excessive sensitivity to handling during blood drawing (Nir and Levy, 1972). In the cited work the feed consumption of the control birds was
REFERENCES Ariyoshi, S., and H. Morimoto, 1956. Studies on the nitrogen metabolism in the fowl. I. Separation of
Downloaded from http://ps.oxfordjournals.org/ at Univ of Iowa-Law Library on June 7, 2015
Assay of FMS activity. The care and treatment of the recipient birds was already described (Nir et al., 1969; Nir and Levy, 1972).
reduced by about 50% and in the treated it was reduced by 75%. According to their effect on the plasma FFA concentration, the different preparations had a similar fat-mobilizing activity in the geese. In cockerels a difference was observed in the activity of the different materials. The FMS obtained from the ad-libitum-fed geese was the most active on fat mobilization (plasma FFA increase) and food intake reduction. The FMS obtained from the starved or overfed geese caused an increase in blood plasma FFA concentration but had no anorexigenic effect. This result is surprising since, as stated by Weil and Stetten (1947) and confirmed in mammals (Chalmers, 1965; Stevenson et al., 1964) and chickens (Nir et al., 1969), the material extracted from fasted animals should be the most active. The marked increase in the amount of FMS excreted during over-feeding (Table 2) agrees well with the observation of Mukaida and Lichton (1971), that FMS 1A excretion by rats was proportional to the amount of food consumed, and of Beaton et al. (1966), that higher FMS 1A was excreted by fed rats compared to fasted rats. It is likely that the geese FMS preparations contain both of the separate anorexigenic and lipid mobilizing substances identified by Beaton et al. (1966). The amount of food intake probably affects the excretion and potency of each and could explain some of the results observed.
632
RESEARCH NOTES
Nir, I., Z. Nitsan and A. Vax, 1973. The influence of force feeding and of protein supplementation to the diet on the metabolizable energy of diets, digestibility of nutrients, nitrogen retention and digestive enzymes output. Ann. Biol. Anim. Bioch. Biophys. 13: 465-479. Nir, I., and M. Perek, 1971. The effect of various protein levels in feed of goslings during the preparatory period on fatty liver production and blood plasma components. Ann. Biol. Anim. Bioch. Biophys. 11: 645-656. Stevenson, J. A. F., B. M. Box and A. J. Szlavko, 1964. A fat-mobilizing and anorectic substance in the urine of fasting rats. Proc. Soc. Exptl. Biol. Med. 115:424-429. Weil, R., and De W. Stetten, Jr., 1947. Urinary excretion of a fat-mobilizing agent. J. Biol. Chem. 168: 129-132.
A THEORETICAL METHOD FOR T H E DETERMINATION O F AMINO ACID A V A I L A B I L I T Y U T I L I Z I N G 14C L A B E L E D P R O T E I N S O U R C E S ' S. V. A M A T O , 2 R. M . FORBES AND H . M . SCOTT
Department of Animal Science, University of Illinois, Urbana, Illinois 61801 (Received for publication October 4, 1974)
ABSTRACT A theoretical method is described whereby the availabilities of amino acids from test proteins are determined relative to the availabilities of amino acids from egg albumen. The method requires that both the test protein and the egg albumen be ,4C labeled. The relative ' availabilities of the amino acids are determined from the change in specific activities of the dietary amino acids and those recovered in the feces. POULTRY SCIENCE 54: 632-634, 1975
INTRODUCTION
A
LTHOUGH the determination of availability of amino acids from protein supplements has been under study for many years, no completely satisfactory method has yet been developed. The most commonly used methods as well as the shortcomings
1. Based on a thesis submitted to the Graduate College of the University of Illinois is partial fulfillment of the requirement for the Ph.D. degree. 2. Present address: Gold Kist Inc., Atlanta, Georgia 30301.
of each have been reviewed by De Muelenaere et al. (1967). In the present communication a theoretical method for the determination of availability of amino acids is described which avoids many of the problems discussed by De Muelenaere et al. (1967). This method is based on the procedure described by Amato (1960) which involves the feeding of a ,4C labeled protein reference standard and l4C labeled test proteins to different groups of animals. The availability of the amino acids of the test proteins is determined relative to that of the reference protein and is based on the
Downloaded from http://ps.oxfordjournals.org/ at Univ of Iowa-Law Library on June 7, 2015
urine for nutritional balance studies. Bull. Natl. Inst. Agr. Sci. Ser. G. 12, 37-44. Beaton, J. R., and A. J. Szlavko and J. A. F. Stevenson, 1966. Factors influencing the excretion of a fat-mobilizing substance in the urine of rats. Can. J. Physiol. Pharmacol. 44: 95-101. Chalmers, T. M., 1965. Lipid-mobilizing activity during fasting. In: Handbook of Physiology. Edited by A. E. Renold and G. F. Cahill, Jr. Waverly Press, Inc., Baltimore, Md. pp. 549-555. Mukaida, C. S., and I. J. Lichton, 1971. Some dietary influences on the excretion and biological activity of an anorexigenic substance in the urine of rats. J. Nutr. 101:767-774. Nir, I., M. K. Dimick and S. Lepkovsky, 1969. A fat-mobilizing substance in chicken urine. Can. J. Physiol. Pharmacol. 47: 435-443. Nir, I., and V. Levy, 1972. Influence of a fat-mobilizing substance from chicken urine on geese. Poultry Sci. 51: 1041-1042.