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Importance of the Cecum in Intestinal Synthesis in the Mature Domestic Fowl*
Importance of the Cecum in Intestinal Synthesis in the Mature Domestic Fowl* J. R. COUCH, H. L. GERMAN, D. R. KNIGHT, PATRICIA SPARKS AND P. B. PEARSON
(Received for publication June 25, 1949)
T
carbohydrates on the microflora of the gastrointestinal tract and reported that dextrin stimulated the development of coliform bacteria and that sucrose depressed the fecal coliforms. These workers reported further that the cecum was the site of the greatest concentration of intestinal microorganisms in the fowl. In a later report Couch, Sunde, Cravens, Elvehjem and Halpin (1949b) observed further that ground whole oats and oat groats supported the intestinal synthesis of biotin in the mature fowl when used to replace either 20 or 40% sucrose in a low biotin diet. The latter workers reported further that approximately two and onehalf times the quantity of biotin ingested by a laying hen fed a practical all-mash diet could be accounted for through excretion in the feces and deposition in the egg.
HE literature on the importance of the ceca in the fowl has been reviewed by Olson and Mann (1935); these portions of the gastrointestinal tract may be removed or occluded with no apparent ill effects. The above workers observed that the pH of this organ was lower than that of either the preceding or succeeding portions of the intestine. Olson and Mann (1935) also reported that there appeared to be a selectivity of materials which gained entrance to the ceca as well as a similar situation with regard to the emptying of these organs. Couch, Cravens, Elvehjem and Halpin (1948) used the laying hen as an experimental animal for studying the intestinal synthesis of biotin as reflected by the biotin content of the egg and by hatchability. These workers reported that dextrin favored the intestinal synthesis of this vitamin but that sucrose, lactose and dried whey did not promote much synthesis. Johansson, Sarles and Shapiro (1948) used the same birds and studied the effect of different
The present investigation was initiated to determine whether the ceca are necessary for the synthesis of B-vitamins in the gastrointestinal tract of the fowl. Criteria used to determine the essentiality of the cecum were of three types. In the first instance the riboflavin, niacin, pantothenic acid, biotin and pteroylglutamic acid content of material taken from the ceca of normal, mature chickens and turkeys which had been maintained under practical range conditions was compared with similar material taken from the in-
* We wish to acknowledge grants of the following as indicated: (1) Pteroylglutamic acid from Lederle Laboratories Division, American Cyanamid Company, Pearl River, N. Y.; (2) All other synthetic vitamins, Merck and Co., Rahway, N. J.; (3) Liver " L " and gelatin, Wilson and Co., Chicago, 111. The data presented in this paper were given at the annual meeting of the American Institute of Nutrition, Detroit, Michigan, April 19-22, 1949. 52
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Departments of Biochemistry and Nutrition, Poultry Husbandry, and Veterinary Pathology and Bacteriology, A &• M College of Texas and Texas Agricultural Experiment Station, College Station
53
CECUM IN INTESTINAL SYNTHESIS
MATERIALS AND METHODS Ten Bronze turkey toms that were approximately nine months of age were selected at random from a flock on range. These birds had been raised under standard recognized management procedures and had been fed a diet of practical feeds that was thought to be complete in all known dietary nutrients. The turkeys were killed and the intestinal tracts removed. The latter were severed just below the duodenum and the ceca were cut off. Contents of the ceca were gently squeezed into a 125 ml. beaker. Likewise the intestinal material below the duodenum, less the ceca, were squeezed into a similar vessel. It was necessary to combine cecal material from three birds in two instances and four in the third case in order to obtain a sufficient quantity on which to determine total dry matter and the five vitamins listed above. Intestinal contents from corresponding birds were combined in a similar manner. Samples of cecal and intestinal material were each thoroughly mixed and aliquots were taken for vitamin determinations and for total dry matter. Ten Single Comb White Leghorn pullets were selected at random from a group on the Poultry Department range, killed, and samples of cecal and intestinal material were collected and analyzed for the five vitamins and total dry matter as outlined above.
At the time the birds described above were killed, other Single Comb White Leghorn pullets were placed in individual laying cages with raised screen bottoms. Five birds of this group were cecectomized prior to being placed in the laying cages. During the following 60 days all hens were fed a practical all-mash diet. At the end of the latter period a 24-hour collection of feces was taken from each of the five cecectomized hens and from a similar number of normal birds that had been kept under the same conditions. The feces from each hen were thoroughly mixed and an aliquot was taken for the determiTABLE 1.—Composition of diets B31
B32B
Sucrose % 63 Starch % 63 Purified casein % 18 18 Gelatin % 5 5 Salts IV % 5 S Liver Fraction " L " % 4 4 Fish oil (3000A-400D) % 2 2 Soybean oil % 3 3 Choline % 0.2 0.2 Oyster shell Ad libitum
B34 63 18 5 5 4 2 3 0.2
All vitamins listed below added ;as indicatedI in mg. per kg. of diet Biotin Thiamin HC1 Riboflavin Ca Pantothenate Niacin 2-methyl-1,4-napthoquinone Pyridoxine HC1 Alpha tocopherol
4 6 IS 100
o.s 4 3
4 6 15 100 0.5 4 3
0.2 4 6 15 100 0.5 4 3
nation of riboflavin, niacin, pantothenic acid, biotin and pteroylglutamic acid. After the feces collection was completed, sixteen normal While Leghorn pullets were selected and divided into four groups of four birds each; a similar group of cecectomized pullets were also selected for the experiment at this time. The management and general experimental procedure was the same as that previously described by Couch et al. (1948).
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testine of these birds. Secondly, the vitamin content (five vitamins listed above) of a 24-hour feces collection from cecectomized pullets was compared with that from normal birds. Finally, the intestinal synthesis of biotin was studied in normal and cecectomized laying hens. Hatchability of fertile eggs and the biotin content of egg yolk were used in evaluating the extent of intestinal synthesis and absorption of biotin.
54
COUCH, GERMAN, KNIGHT, SPARKS AND PEARSON
was used for the determination of riboflavin, of Neal and Strong (1943) for the determination of pantothenic acid, of Krehl, Strong and Elvehjem (1943) for the determination of niacin. The media of Flynn (1948) was used for the determination of pteroylglutamic acid with Lactobacillus casei as the test organism. The titrimetric procedure was used to measure the growth of the organism; PGA was liberated with chick pancreas in phosphate buffer at pH 7.0.
TABLE 2.—Vitamin content of cecal and intestinal material of mature turkeys and chickens Turkeys
Chickens
Source of Material Ceca Vitamin
Intestine
Micrograms per gram (dry weight)
Ceca
Intestine
Micrograms per gram (dry weight)
59.4 (48.0-75.3)
27.5 (23.2-35.9)
74.3 (58.4-96.6)
28.7 (22.9-33.2)
255.3 (222.5-309.9)
185.7 (147.5-249.7)
392.9 (338.1-491.5)
212.3 (193.6-222.2)
Pantothenic acid
13.6 (13.2-14.0)
12.6 (10.4-14.9)
96.1 (80.7-111.5)
25.5 (13.8^3.6)
Folic acid
20.5 (17.8-24.3)
3 3 (2.7^.0)
32.8 (20.6-49.3)
4.2 (3.1-5.3)
Biotin
11.2 (10.4-12.7)
1.3 (1.0-1.8)
21.1 (14.2-31.9)
1.4 (1.1-1.7)
Riboflavin Niacin
The effect of removal of the cecum on the intestinal synthesis of biotin was studied by using starch as a source of carbohydrate in a low biotin diet (B32B) and feeding this diet to one group of normal hens and likewise to a comparable group of cecectomized birds. A group which was fed sucrose as a source of carbohydrate in a low biotin diet (B31) served as the negative control while a similar one to which biotin had been added was the positive control. An additional group of four pullets was fed a practical all-mash diet similar in composition to that used earlier by Couch and coworkers (1948). The method of Snell and Strong (1939)
RESULTS AND DISCUSSION
This investigation has shown that cecal material taken from the turkey and chicken under practical conditions contains a higher content of riboflavin, niacin, pantothenic acid, PGA and biotin than does the material taken from the remainder of the intestine below the duodenum. These differences are somewhat greater in the case of the White Leghorn pullets than the Broad Breasted Bronze turkey toms (Table 2). Cecal material of the turkey contained over two times as much riboflavin, somewhat more niacin, about the same amount of pantothenic acid, six times as much PGA and nine times as much biotin as
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The composition of the diets is shown in Table 1. Crystalline B-vitamins with the exception of biotin, pteroylglutamic acid and vitamin B12 were added in amounts which were thought to meet the requirements of the birds. Liver Fraction " L " was used as a source of PGA, vitamin B ]2 and unidentified factors. Biotin was added to diet B34. Fish oil was used as a source of vitamins A and D. Vitamins E and K were supplied as indicated in Table 1.
CECUM IN INTESTINAL SYNTHESIS
did intestinal material from these birds (Table 2). The material taken from the ceca of the White Leghorn pullets contained over two and one-half times as much riboflavin, almost twice as much niacin, practically four times as much pantothenic acid, about eight times as
Source of Material Vitamin
Normal hen
Cecectomized Hen
Micrograms per gram (dry weight)
Pantothenic Acid
38.0 (10.0-58.0)
31.0 (15.0-75.0)
Niacin
70.0 (58.0-90.0)
90.0 (61.0-150.0)
Riboflavin
6.0 (4.0-7.0)
5.0 (3.0-6.0)
Folic Acid
2.4 (0.9-3.8)
2.5 (2.0-3.1)
0.66 (0.50-0.77)
0.64 (0.52-0.71)
Biotin
much PGA and fifteen times as much biotin as intestinal material taken from the same fowls (Table 2). From these data it is apparent that the cecal contents of the turkey and chicken have a higher value for the five vitamins mentioned above than does intestinal material taken from the same birds. The explanation for this observation may be that food material remains in the cecum a longer period of time thus bringing about more complete digestion possibly through bacterial action and fermentation. Johansson, Sarles and Shapiro (1948) found that the cecum of the fowl was the site of the greatest concentration of intestinal microorganisms and reported further that cecal pouch contents were usually evacuated in the morning and evening. Olson and Mann (1935) have shown that the ceca of the
fowl empties intermittently and may hold some types of material for considerable periods of time. When normal and cecectomized birds were fed a practical all-mash diet and maintained in individual laying cages with raised screen bottoms it was observed that the riboflavin, niacin, pantothenic acid, PGA and biotin content of a 24-hour feces collection from the two types of pullets were not appreciably different (Table 3). These data indicate that the ceca are apparently not necessary for synthesis of B-vitamins in the gastrointestinal tract of the fowl. It is realized in this case that the cecectomized pullets had a 60-day period in which to adjust the microflora of the tract after removal of the ceca and prior to collection of the feces samples (Table 3). It should be mentioned that earlier studies have shown that the ceca of the fowl could be removed or occluded without any apparent ill effects (Olson and Mann), (1935). The cecum was not required for the intestinal synthesis of biotin in the mature fowl (Figures 1 and 2) when starch was used as a source of carbohydrate (diet B32B). The feeding of sucrose in a low biotin diet to normal hens gave results similar to those reported earlier by Couch et al. (1948, 1949b). The percent hatchability in this case decreased to less than 10% by the end of the second week and to zero by the end of the fourth week. Live chicks were not obtained from hens fed diet B31 (sucrose) after the fourth week. When this diet was supplemented with biotin (diet B34) the percent hatchability was about the same as that observed in eggs from hens fed a practical all-mash diet. The percent hatchability of eggs from normal hens fed starch (diet B32B) is unquestionably higher than that of those fed sucrose (Figure 1). This is in line with earlier reports by Couch,
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TABLE 3.—Vitamin content of a twenty-four hour collection of fecal material from normal and cecectomized laying hens
55
56
COUCH, GERMAN, KNIGHT, SPARKS AND PEARSON
Cravens, Elvehjem and Halpin (1948) and Couch, et al. (1949b). However, it is interesting to note that the percent hatchability of eggs from cecectomized hens fed diet B32B is consistently higher than that of similar normal birds fed the same
higher percentage hatchability obtained in eggs from hens with the ceca removed as compared with the normals when both were fed starch in a low biotin diet might be explained on the basis that the removal of the ceca might have de-
/ /
\B32B (NORMAL)
B-31 5
6 WEEKS FIG. 1. Effect on hatchability of feeding a low biotin diet to cecectomized and normal hens: diet B31, sucrose; diet B32B, starch; diet B34, biotin.
diet (Figure 1). Johannson, Sarles and Shapiro (1948) reported that dextrin favored the growth of biotin-synthesizing organisms (coliforms) while sucrose favored the growth of biotin utilizing organisms (yeasts). It is possible that the
creased the number of biotin utilizing organisms in the tract of the cecectomized birds. The feeding of starch to normal and cecectomized hens promoted the intestinal synthesis of biotin as measured by the
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PRACTICAL
CECUM IN INTESTINAL SYNTHESIS
biotin content of the egg yolk (Figure 2). The biotin is reported as micrograms per gram of egg yolk on a fresh basis. The biotin content of egg yolks from hens fed sucrose (B31) decreased in almost a linear relationship until the end of the second
57
der similar conditions (Couch, Cravens, Elvehjem and Halpin, 1949a). It may be noted that the biotin values of egg yolk from hens fed diet B34 are higher than those from hens fed the practical all-mash diet. The differences in the biotin content
PRACTICAL
(CECECTOMIZED) .^B32B_ __B-32B (NORMAL) ;_.__.-.•==*^>* B3I
"*•
ZlI
S
2
3
4
9
WEEKS FIG. 2. Effect on the biotin content of egg yolk of feeding a low biotin diet to cecectomized and normal hens: diet B31, sucrose; diet B32B, starch; diet B34, biotin.
week and remained fairly constant at a low level after this time. When the sucrose diet was supplemented with biotin (B34) there was an initial decrease in the quantity of the vitamin per gram of egg yolk. This initial decrease has been observed before un-
of egg yolk from cecectomized hens fed starch (diet B32B) and those from normal birds fed this carbohydrate are not so striking as was true with the percent hatchability as discussed above. As a matter of fact, egg yolk from the normal
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B34
58
COUCH, GERMAN, KNIGHT, SPARKS AND PEARSON
hens fed starch (B32B) contained a somewhat larger quantity of biotin than that from the cecectomized birds at the end of the second and third weeks while the reverse is true for values obtained at the end of the fourth and ninth weeks. SUMMARY
ADDENDUM
During the time when this manuscript was being prepared the authors received a private communication from Dr. W. W. Cravens, Department of Poultry Hus-
REFERENCES Couch, J. R., W. W. Cravens, C. A. Elvehjem and J. G. Halpin 1948. Relation of carbohydrate to intestinal synthesis of biotin and hatchability in mature fowl. J. Nutr. 35: 57-72. Couch, J. R., W. W. Cravens, C. A. Elvehjem and J. G. Halpin 1949a. Studies on the function of biotin in the domestic fowl. Arch. Biochem. 21: 77-86. Couch, J. R., M. L. Sunde, W. W. Cravens, C. A. Elvehjem and J. G. Halpin 1949b. Effect of oats, oat products and fat on the intestinal synthesis of biotin in mature fowl. J. Nutr. 37: 251-262. Flynn, Laura M., 1948. Private communication. Johansson, K. R., W. B. Sarles,. and S. K. Shapiro, 1948 The intestinal microflora of hens as influenced by various carbohydrates in a biotindeficient ration. J. Bact. 56: 619-634. Krehl, W. A., F. M. Strong and C. A. Elvehjem, 1943. Determination of nicotinic acid. Ind. Eng. Chem. Anal. Ed. 15:471-175. Neal, A. L., and Strong, F. M., 1943. Microbiological determination of pantothenic acid. Ind. Eng. Chem. Anal. Ed. 15: 654-657. Olson, Carl, Jr., and F. C. Mann, 1935. The physiology of the cecum of the domestic fowl. J. Am. Vet. Med. Assn. 87: 151-159. Snell, E. E., and F. M. Strong, 1939. A microbiological assay for riboflavin. Ind. Eng. Chem. Anal. Ed. 11:346-350. Wright, L. D., and Skeggs, H. R., 1944. Determination of biotin with Lactobacillus arabinosus. Proc. Soc. Exp. Biol. Med. 56: 95-98.
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Cecal material taken from mature chickens and turkeys maintained on range and fed practical diets contained a higher concentration of riboflavin, niacin, pantothenic acid, biotin and PGA than did intestinal material taken from the same birds. Fecal material from cecectomized birds contained approximately the same content of the five B-vitamins listed above as did a similar collection from normal birds. The cecum is not required for the synthesis of biotin when mature hens are fed starch as a source of carbohydrate in a low biotin diet. An indication was obtained to the effect that removal of the cecum enhanced the intestinal synthesis of biotin in the gastrointestinal tract of the laying domestic fowl.
bandry, University of Wisconsin, Madison, Wisconsin, to the effect that the Wisconsin workers had obtained data which confirm results presented herein.
(Received for publication June 25, 1949)
T
carbohydrates on the microflora of the gastrointestinal tract and reported that dextrin stimulated the development of coliform bacteria and that sucrose depressed the fecal coliforms. These workers reported further that the cecum was the site of the greatest concentration of intestinal microorganisms in the fowl. In a later report Couch, Sunde, Cravens, Elvehjem and Halpin (1949b) observed further that ground whole oats and oat groats supported the intestinal synthesis of biotin in the mature fowl when used to replace either 20 or 40% sucrose in a low biotin diet. The latter workers reported further that approximately two and onehalf times the quantity of biotin ingested by a laying hen fed a practical all-mash diet could be accounted for through excretion in the feces and deposition in the egg.
HE literature on the importance of the ceca in the fowl has been reviewed by Olson and Mann (1935); these portions of the gastrointestinal tract may be removed or occluded with no apparent ill effects. The above workers observed that the pH of this organ was lower than that of either the preceding or succeeding portions of the intestine. Olson and Mann (1935) also reported that there appeared to be a selectivity of materials which gained entrance to the ceca as well as a similar situation with regard to the emptying of these organs. Couch, Cravens, Elvehjem and Halpin (1948) used the laying hen as an experimental animal for studying the intestinal synthesis of biotin as reflected by the biotin content of the egg and by hatchability. These workers reported that dextrin favored the intestinal synthesis of this vitamin but that sucrose, lactose and dried whey did not promote much synthesis. Johansson, Sarles and Shapiro (1948) used the same birds and studied the effect of different
The present investigation was initiated to determine whether the ceca are necessary for the synthesis of B-vitamins in the gastrointestinal tract of the fowl. Criteria used to determine the essentiality of the cecum were of three types. In the first instance the riboflavin, niacin, pantothenic acid, biotin and pteroylglutamic acid content of material taken from the ceca of normal, mature chickens and turkeys which had been maintained under practical range conditions was compared with similar material taken from the in-
* We wish to acknowledge grants of the following as indicated: (1) Pteroylglutamic acid from Lederle Laboratories Division, American Cyanamid Company, Pearl River, N. Y.; (2) All other synthetic vitamins, Merck and Co., Rahway, N. J.; (3) Liver " L " and gelatin, Wilson and Co., Chicago, 111. The data presented in this paper were given at the annual meeting of the American Institute of Nutrition, Detroit, Michigan, April 19-22, 1949. 52
Downloaded from http://ps.oxfordjournals.org/ at National Institute of Education Library, Serials Unit on May 20, 2015
Departments of Biochemistry and Nutrition, Poultry Husbandry, and Veterinary Pathology and Bacteriology, A &• M College of Texas and Texas Agricultural Experiment Station, College Station
53
CECUM IN INTESTINAL SYNTHESIS
MATERIALS AND METHODS Ten Bronze turkey toms that were approximately nine months of age were selected at random from a flock on range. These birds had been raised under standard recognized management procedures and had been fed a diet of practical feeds that was thought to be complete in all known dietary nutrients. The turkeys were killed and the intestinal tracts removed. The latter were severed just below the duodenum and the ceca were cut off. Contents of the ceca were gently squeezed into a 125 ml. beaker. Likewise the intestinal material below the duodenum, less the ceca, were squeezed into a similar vessel. It was necessary to combine cecal material from three birds in two instances and four in the third case in order to obtain a sufficient quantity on which to determine total dry matter and the five vitamins listed above. Intestinal contents from corresponding birds were combined in a similar manner. Samples of cecal and intestinal material were each thoroughly mixed and aliquots were taken for vitamin determinations and for total dry matter. Ten Single Comb White Leghorn pullets were selected at random from a group on the Poultry Department range, killed, and samples of cecal and intestinal material were collected and analyzed for the five vitamins and total dry matter as outlined above.
At the time the birds described above were killed, other Single Comb White Leghorn pullets were placed in individual laying cages with raised screen bottoms. Five birds of this group were cecectomized prior to being placed in the laying cages. During the following 60 days all hens were fed a practical all-mash diet. At the end of the latter period a 24-hour collection of feces was taken from each of the five cecectomized hens and from a similar number of normal birds that had been kept under the same conditions. The feces from each hen were thoroughly mixed and an aliquot was taken for the determiTABLE 1.—Composition of diets B31
B32B
Sucrose % 63 Starch % 63 Purified casein % 18 18 Gelatin % 5 5 Salts IV % 5 S Liver Fraction " L " % 4 4 Fish oil (3000A-400D) % 2 2 Soybean oil % 3 3 Choline % 0.2 0.2 Oyster shell Ad libitum
B34 63 18 5 5 4 2 3 0.2
All vitamins listed below added ;as indicatedI in mg. per kg. of diet Biotin Thiamin HC1 Riboflavin Ca Pantothenate Niacin 2-methyl-1,4-napthoquinone Pyridoxine HC1 Alpha tocopherol
4 6 IS 100
o.s 4 3
4 6 15 100 0.5 4 3
0.2 4 6 15 100 0.5 4 3
nation of riboflavin, niacin, pantothenic acid, biotin and pteroylglutamic acid. After the feces collection was completed, sixteen normal While Leghorn pullets were selected and divided into four groups of four birds each; a similar group of cecectomized pullets were also selected for the experiment at this time. The management and general experimental procedure was the same as that previously described by Couch et al. (1948).
Downloaded from http://ps.oxfordjournals.org/ at National Institute of Education Library, Serials Unit on May 20, 2015
testine of these birds. Secondly, the vitamin content (five vitamins listed above) of a 24-hour feces collection from cecectomized pullets was compared with that from normal birds. Finally, the intestinal synthesis of biotin was studied in normal and cecectomized laying hens. Hatchability of fertile eggs and the biotin content of egg yolk were used in evaluating the extent of intestinal synthesis and absorption of biotin.
54
COUCH, GERMAN, KNIGHT, SPARKS AND PEARSON
was used for the determination of riboflavin, of Neal and Strong (1943) for the determination of pantothenic acid, of Krehl, Strong and Elvehjem (1943) for the determination of niacin. The media of Flynn (1948) was used for the determination of pteroylglutamic acid with Lactobacillus casei as the test organism. The titrimetric procedure was used to measure the growth of the organism; PGA was liberated with chick pancreas in phosphate buffer at pH 7.0.
TABLE 2.—Vitamin content of cecal and intestinal material of mature turkeys and chickens Turkeys
Chickens
Source of Material Ceca Vitamin
Intestine
Micrograms per gram (dry weight)
Ceca
Intestine
Micrograms per gram (dry weight)
59.4 (48.0-75.3)
27.5 (23.2-35.9)
74.3 (58.4-96.6)
28.7 (22.9-33.2)
255.3 (222.5-309.9)
185.7 (147.5-249.7)
392.9 (338.1-491.5)
212.3 (193.6-222.2)
Pantothenic acid
13.6 (13.2-14.0)
12.6 (10.4-14.9)
96.1 (80.7-111.5)
25.5 (13.8^3.6)
Folic acid
20.5 (17.8-24.3)
3 3 (2.7^.0)
32.8 (20.6-49.3)
4.2 (3.1-5.3)
Biotin
11.2 (10.4-12.7)
1.3 (1.0-1.8)
21.1 (14.2-31.9)
1.4 (1.1-1.7)
Riboflavin Niacin
The effect of removal of the cecum on the intestinal synthesis of biotin was studied by using starch as a source of carbohydrate in a low biotin diet (B32B) and feeding this diet to one group of normal hens and likewise to a comparable group of cecectomized birds. A group which was fed sucrose as a source of carbohydrate in a low biotin diet (B31) served as the negative control while a similar one to which biotin had been added was the positive control. An additional group of four pullets was fed a practical all-mash diet similar in composition to that used earlier by Couch and coworkers (1948). The method of Snell and Strong (1939)
RESULTS AND DISCUSSION
This investigation has shown that cecal material taken from the turkey and chicken under practical conditions contains a higher content of riboflavin, niacin, pantothenic acid, PGA and biotin than does the material taken from the remainder of the intestine below the duodenum. These differences are somewhat greater in the case of the White Leghorn pullets than the Broad Breasted Bronze turkey toms (Table 2). Cecal material of the turkey contained over two times as much riboflavin, somewhat more niacin, about the same amount of pantothenic acid, six times as much PGA and nine times as much biotin as
Downloaded from http://ps.oxfordjournals.org/ at National Institute of Education Library, Serials Unit on May 20, 2015
The composition of the diets is shown in Table 1. Crystalline B-vitamins with the exception of biotin, pteroylglutamic acid and vitamin B12 were added in amounts which were thought to meet the requirements of the birds. Liver Fraction " L " was used as a source of PGA, vitamin B ]2 and unidentified factors. Biotin was added to diet B34. Fish oil was used as a source of vitamins A and D. Vitamins E and K were supplied as indicated in Table 1.
CECUM IN INTESTINAL SYNTHESIS
did intestinal material from these birds (Table 2). The material taken from the ceca of the White Leghorn pullets contained over two and one-half times as much riboflavin, almost twice as much niacin, practically four times as much pantothenic acid, about eight times as
Source of Material Vitamin
Normal hen
Cecectomized Hen
Micrograms per gram (dry weight)
Pantothenic Acid
38.0 (10.0-58.0)
31.0 (15.0-75.0)
Niacin
70.0 (58.0-90.0)
90.0 (61.0-150.0)
Riboflavin
6.0 (4.0-7.0)
5.0 (3.0-6.0)
Folic Acid
2.4 (0.9-3.8)
2.5 (2.0-3.1)
0.66 (0.50-0.77)
0.64 (0.52-0.71)
Biotin
much PGA and fifteen times as much biotin as intestinal material taken from the same fowls (Table 2). From these data it is apparent that the cecal contents of the turkey and chicken have a higher value for the five vitamins mentioned above than does intestinal material taken from the same birds. The explanation for this observation may be that food material remains in the cecum a longer period of time thus bringing about more complete digestion possibly through bacterial action and fermentation. Johansson, Sarles and Shapiro (1948) found that the cecum of the fowl was the site of the greatest concentration of intestinal microorganisms and reported further that cecal pouch contents were usually evacuated in the morning and evening. Olson and Mann (1935) have shown that the ceca of the
fowl empties intermittently and may hold some types of material for considerable periods of time. When normal and cecectomized birds were fed a practical all-mash diet and maintained in individual laying cages with raised screen bottoms it was observed that the riboflavin, niacin, pantothenic acid, PGA and biotin content of a 24-hour feces collection from the two types of pullets were not appreciably different (Table 3). These data indicate that the ceca are apparently not necessary for synthesis of B-vitamins in the gastrointestinal tract of the fowl. It is realized in this case that the cecectomized pullets had a 60-day period in which to adjust the microflora of the tract after removal of the ceca and prior to collection of the feces samples (Table 3). It should be mentioned that earlier studies have shown that the ceca of the fowl could be removed or occluded without any apparent ill effects (Olson and Mann), (1935). The cecum was not required for the intestinal synthesis of biotin in the mature fowl (Figures 1 and 2) when starch was used as a source of carbohydrate (diet B32B). The feeding of sucrose in a low biotin diet to normal hens gave results similar to those reported earlier by Couch et al. (1948, 1949b). The percent hatchability in this case decreased to less than 10% by the end of the second week and to zero by the end of the fourth week. Live chicks were not obtained from hens fed diet B31 (sucrose) after the fourth week. When this diet was supplemented with biotin (diet B34) the percent hatchability was about the same as that observed in eggs from hens fed a practical all-mash diet. The percent hatchability of eggs from normal hens fed starch (diet B32B) is unquestionably higher than that of those fed sucrose (Figure 1). This is in line with earlier reports by Couch,
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TABLE 3.—Vitamin content of a twenty-four hour collection of fecal material from normal and cecectomized laying hens
55
56
COUCH, GERMAN, KNIGHT, SPARKS AND PEARSON
Cravens, Elvehjem and Halpin (1948) and Couch, et al. (1949b). However, it is interesting to note that the percent hatchability of eggs from cecectomized hens fed diet B32B is consistently higher than that of similar normal birds fed the same
higher percentage hatchability obtained in eggs from hens with the ceca removed as compared with the normals when both were fed starch in a low biotin diet might be explained on the basis that the removal of the ceca might have de-
/ /
\B32B (NORMAL)
B-31 5
6 WEEKS FIG. 1. Effect on hatchability of feeding a low biotin diet to cecectomized and normal hens: diet B31, sucrose; diet B32B, starch; diet B34, biotin.
diet (Figure 1). Johannson, Sarles and Shapiro (1948) reported that dextrin favored the growth of biotin-synthesizing organisms (coliforms) while sucrose favored the growth of biotin utilizing organisms (yeasts). It is possible that the
creased the number of biotin utilizing organisms in the tract of the cecectomized birds. The feeding of starch to normal and cecectomized hens promoted the intestinal synthesis of biotin as measured by the
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PRACTICAL
CECUM IN INTESTINAL SYNTHESIS
biotin content of the egg yolk (Figure 2). The biotin is reported as micrograms per gram of egg yolk on a fresh basis. The biotin content of egg yolks from hens fed sucrose (B31) decreased in almost a linear relationship until the end of the second
57
der similar conditions (Couch, Cravens, Elvehjem and Halpin, 1949a). It may be noted that the biotin values of egg yolk from hens fed diet B34 are higher than those from hens fed the practical all-mash diet. The differences in the biotin content
PRACTICAL
(CECECTOMIZED) .^B32B_ __B-32B (NORMAL) ;_.__.-.•==*^>* B3I
"*•
ZlI
S
2
3
4
9
WEEKS FIG. 2. Effect on the biotin content of egg yolk of feeding a low biotin diet to cecectomized and normal hens: diet B31, sucrose; diet B32B, starch; diet B34, biotin.
week and remained fairly constant at a low level after this time. When the sucrose diet was supplemented with biotin (B34) there was an initial decrease in the quantity of the vitamin per gram of egg yolk. This initial decrease has been observed before un-
of egg yolk from cecectomized hens fed starch (diet B32B) and those from normal birds fed this carbohydrate are not so striking as was true with the percent hatchability as discussed above. As a matter of fact, egg yolk from the normal
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B34
58
COUCH, GERMAN, KNIGHT, SPARKS AND PEARSON
hens fed starch (B32B) contained a somewhat larger quantity of biotin than that from the cecectomized birds at the end of the second and third weeks while the reverse is true for values obtained at the end of the fourth and ninth weeks. SUMMARY
ADDENDUM
During the time when this manuscript was being prepared the authors received a private communication from Dr. W. W. Cravens, Department of Poultry Hus-
REFERENCES Couch, J. R., W. W. Cravens, C. A. Elvehjem and J. G. Halpin 1948. Relation of carbohydrate to intestinal synthesis of biotin and hatchability in mature fowl. J. Nutr. 35: 57-72. Couch, J. R., W. W. Cravens, C. A. Elvehjem and J. G. Halpin 1949a. Studies on the function of biotin in the domestic fowl. Arch. Biochem. 21: 77-86. Couch, J. R., M. L. Sunde, W. W. Cravens, C. A. Elvehjem and J. G. Halpin 1949b. Effect of oats, oat products and fat on the intestinal synthesis of biotin in mature fowl. J. Nutr. 37: 251-262. Flynn, Laura M., 1948. Private communication. Johansson, K. R., W. B. Sarles,. and S. K. Shapiro, 1948 The intestinal microflora of hens as influenced by various carbohydrates in a biotindeficient ration. J. Bact. 56: 619-634. Krehl, W. A., F. M. Strong and C. A. Elvehjem, 1943. Determination of nicotinic acid. Ind. Eng. Chem. Anal. Ed. 15:471-175. Neal, A. L., and Strong, F. M., 1943. Microbiological determination of pantothenic acid. Ind. Eng. Chem. Anal. Ed. 15: 654-657. Olson, Carl, Jr., and F. C. Mann, 1935. The physiology of the cecum of the domestic fowl. J. Am. Vet. Med. Assn. 87: 151-159. Snell, E. E., and F. M. Strong, 1939. A microbiological assay for riboflavin. Ind. Eng. Chem. Anal. Ed. 11:346-350. Wright, L. D., and Skeggs, H. R., 1944. Determination of biotin with Lactobacillus arabinosus. Proc. Soc. Exp. Biol. Med. 56: 95-98.
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Cecal material taken from mature chickens and turkeys maintained on range and fed practical diets contained a higher concentration of riboflavin, niacin, pantothenic acid, biotin and PGA than did intestinal material taken from the same birds. Fecal material from cecectomized birds contained approximately the same content of the five B-vitamins listed above as did a similar collection from normal birds. The cecum is not required for the synthesis of biotin when mature hens are fed starch as a source of carbohydrate in a low biotin diet. An indication was obtained to the effect that removal of the cecum enhanced the intestinal synthesis of biotin in the gastrointestinal tract of the laying domestic fowl.
bandry, University of Wisconsin, Madison, Wisconsin, to the effect that the Wisconsin workers had obtained data which confirm results presented herein.