- Email: [email protected]
Essential Fatty Acid Deficiency in the Laying Hen1
1014
A. W. KOTULA, J. E. THOMSON, J. F. NOVOTNY AND E. H. MCNALLY
score for unfrozen thighs, and a 1.3 increase in score for frozen thighs. ACKNOWLEDGMENTS
Essential Fatty Acid Deficiency in the Laying Hen 1 LEO S. JENSEN AND JOHN V. SHUTZE Department of Poultry Science, Washington State University, Pullman (Received for publication February 11, 1963)
AN ESSENTIAL fatty acid deficiency - i * - in the rat has been thoroughly investigated and the symptoms are well defined (Aase-Jorgensen, 1961). Less study has been devoted to such a deficiency in birds. An essential fatty acid deficiency in growing chickens was reported by Bieri et al. (1956) in which chicks fed a fat-free diet grew less after 6 weeks of age than those fed the basal diet supplemented with 4% corn oil. No marked deficiency symptoms were observed, however, even after 20 weeks of depletion. Machlin and Gordon (1961) obtained growth responses to linoleic acid supplements during the first two weeks of the chick's life. The growth depression on the fat-free diet was accentuated by adding saturated fatty acids
1
Scientific Paper No. 2312, Washington Agricultural Experiment Stations, Pullman. Project 1532. Supported in part by funds from Medical and Biological Research by State of Washington Initiative No. 171.
and cholesterol. Fatty acid analysis of heart, testes and cerebrum revealed an increase in triene and decrease in tetraene acids in the deficient birds. Dam et al. (1956) and Bieri et al. (1957) also observed a similar change. An increased ratio of triene to tetraene was orignally observed as a result of an essential fatty acid deficiency in other species, and Holman (1960) has proposed the determination of this ratio in tissues as a criterion for estimating the essential fatty acid requirement in animals. Reiser (1950, 1951) studied an essential fatty acid deficiency in the laying hen and reported that when very low levels of linoleic acid were obtained in eggs, the reproductive performance of the hens apparently was not affected. An earlier conclusion that the hen can synthesize linoleic acid de novo was later reversed by Murty et al. (I960), using more refined techniques. In a previous report from our labora-
Downloaded from http://ps.oxfordjournals.org/ at OhioLINK Offices on May 2, 2015
Appreciation is expressed for the cooperation of the Institute of American Poultry Industries which supplied the chickens; Poultry Nutrition Investigations, Poultry Research Branch, ARS, which prepared diets and maintained the birds; and E. James Koch, ARS, U. S. Department of Agriculture, for advice and assistance in analyzing the data.
REFERENCES Brant, A. W., and G. F. Stewart, 1950. Bone darkening in frozen poultry. Food Tech. 4:168-174. Duncan, D. B., 1955. Multiple range and multiple F tests. Biometrics, 11: 1-7. Ellis, C , and J. G. Woodroof, 1959. Prevention of bone darkening. Food Tech. 13: 533-538. Essary, E. 0., and E. L. Wisman, 1962. Personal communication. Koonz, C. H., 1946. Poultry and Eggs. Refrig. Eng. Data Book 59. Woodroof, J. G., and E. Shelor, 1948. Prevention of bone darkening in frozen-packed chickens. Food Industries, 20: 48-52.
1015
FATTY ACID DEFICIENCY
tory, corn oil was shown to increase egg weight of birds fed a semi-purified diet (Jensen et al., 1958). Hatchability was also shown to be adversely affected after some time on the semi-purified diet unless corn was included. Linoleic acid was found to be the major factor in corn oil responsible for the egg weight increase (Shutze and Jensen, 1963). The present experiments were undertaken to study further an essential fatty acid deficiency in the laying hen.
Single Comb White Leghorn chicks were reared on three different diets from day of age to maturity and were then fed similar diets during approximately ten months of egg production. The chicks were brooded in battery brooders with raised wire floors and were then transferred to wire floored colony cages with about 16 birds per pen. At 20 weeks of age the birds were moved to individual hen cages. Feed and water were supplied ad libitum. In order to obtain fertile eggs, the hens were artificially inseminated with semen from males maintained on a practical breeder diet. Chicks hatched from eggs laid by the hens fed the three diets were reared to maturity and kept for 39 weeks in production. Composition of the basal low fat and practical laying diet is shown in Table 1. Rearing diets were similar, except that level of protein was higher for the first 8 weeks and calcium carbonate was completely removed. No linoleic acid was detected in the low fat basal diet by gasliquid chromatography, whereas the low fat diet supplemented with safflower oil and the practical diet contained considerable linoleic acid. All unbroken eggs laid were weighed at two week intervals. Samples of eggs from each treatment were incubated to deter-
Layer diets Ingredients Low Fat Practical Corn, yellow Cerelose Defatted fish meal1 Soybean meal, dehulled Fish meal, herring Dehydrated alfalfa meal Dried brewers' yeast Limestone Dicalcium phosphate NaCl, iodized Manganese sulfate KC1 Trace mineral mixture Vitamin mixture
% 62.80 30.00
— — —
3.00 3.20
—
0.40
—
0.40 0.05 2 1.003
%
74.75
— —
8.00 5.00 3.00 2.00 3.25 3.00 0.50 0.02
— —
0.50"
1 Obtained from Viobin Corporation, Monticello, Illinois. 2 Supplied the following p.p.m.: Mn, 50.0; Fe, 50.0; Cu, 5.0; Co, 0.5; I, 1.5; Zn, 50.0. 3 Supplied the following per kg. of diet: vitamin A, 8,811 I.U.; vitamin D 3 , 881 I.U.; menadione sodium bisulfite, 6.6. mg.; vitamin E, 11.0 I.U.; riboflavin, 4.4. mg.; pyridoxine, 4.4 mg.; Ca-pantothenate, 13.9 mg.; niacin, 22.0 mg.; folic acid, 0.44 mg.; B12, 2.2 meg.; ethoxyquin, 249.; bacitracin, 11 mg. 1 Supplied the following per kg. of diet: vitamin A, 6,6101.U.; vitamin D 3 , 7441.U.; riboflavin, 4.4. mg.; vitamin E, 22.0 I.U.; Pn2, 9.9 meg.; ethoxyquin, 249 mg.; bacitracin, 11.0 mg. Note: The safflower diet consisted of adding 5% safflower oil to the low fat diet at the expense of cerelose for the first experiment. During the second experiment, 11.2% of the cerelose was left out and only 5% safflower oil was added in order to make these two diets isocaloric.
mine the effect on fertility and hatchability. Fatty acids were determined by a gas-liquid chromatography method as follows: the dried feed samples and lyophilized egg samples were extracted for 12 hours in a Soxhlet with a 2:1 mixture of chloroform and methanol. The carcasses were ground through a meat grinder and thoroughly mixed before an aliquot was taken for lyophilization and extraction. Methyl esters were formed by interesterification, using sodium methoxide as a catalyst (Craig and Murty, 1959). A Beckman GC2A model instrument with a thermol conductivity detector was used for analysis of the methyl esters. A | in.
Downloaded from http://ps.oxfordjournals.org/ at OhioLINK Offices on May 2, 2015
EXPERIMENTAL
TABLE 1.—Composition of diets
1016
L. S. J E N S E N AND J. V. SHUTZE
TABLE 2.—EJfect of an essential fatty acid deficiency on egg production % Average egg production Dlet
First generation 1
Second generation 2
Low fat basal Low fat basal + 5 % safflower oil Practical
44.6
29.1
45.3 56.5
38.0 56.7
1 1
Average for 42 week period. Average for 39 week period.
RESULTS
Although the chicks fed the basal diet supplemented with safflower oil grew faster t h a n the chicks fed the unsupplemented basal diet, both groups reached sexual maturity at about the same age. Chicks fed the practical diet grew at about the same rate as those on the unsupplemented basal diet. Similar observations were made with the second generation chicks. There was no difference in r a t e of lay between pullets fed the basal diet and the diet supplemented with safflower oil during the first experiment (Table 2). However, birds fed the practical diet laid at a faster rate. In the second experiment (involving off-spring from the initial birds used in the study), slightly lower egg production was obtained with the unsupplemented basal diet. Again, birds fed the practical diet laid a t a more rapid rate, suggesting t h a t factors other t h a n essential fatty acids were influencing egg production. Egg weight was markedly influenced by the diet. Average egg weight of birds fed the diet supplemented with safflower oil was consistently 2-6 grams heavier t h a n
DISCUSSION Essential fatty acid deficiency in the laying hen resulted in a marked reduction in egg weight, a depression in hatchability of fertile eggs, but had little effect on rate of egg production and apparently no TABLE 3.—Effect of an essential fatly acid deficiency in hens on fertility and hatchability Diet
Month of production
First generation Low fat basal Low fat basal plus safflower oil Practical Low fat basal Low fat basal plus safflower oil Practical Low fat basal Low fat basal plus safflower oil Practical Low fat basal Low fat basal plus safflower oil Practical Second generation Low fat basal Low fat basal plus safflower oil Practical
No. eggs set
Fertility
% Hatch of fertile eggs
%
4
111
73.6
38.3
4 4
178 215
49.4 46.5
62.5 72.0
S
111
72.5
33.7
5 5
137 125
84.7 87.2
68.0 86.3
7
138
79.0
56.8
7 7
212 180
83.0 73.3
68.2 84.1
8
144
66.6
79.3
8 8
195 281
79.0 80.0
81.3 78.7
6-7
318
74.2
61.0
6-7 6-7
508 744
73.6 88.3
71.1 84.8
•
Downloaded from http://ps.oxfordjournals.org/ at OhioLINK Offices on May 2, 2015
X12 ft. copper column was packed with 2 5 % diethylene glycol succinate absorbed on 40-60 mesh acid washed chromosorb W and helium gas was used as a carrier. Column temperature was maintained at 235° C.
that of birds fed the unsupplemented basal diet (Figs. 1 and 2). Eggs from hens fed the practical diet were consistently heavier than both the groups fed the purified diets, indicating t h a t factors other than essential fatty acids were also influencing egg weight. No consistent effect of diet on fertility was observed (Table 3), but hatchability of fertile eggs was increased by supplementing the purified diet with safflower oil or by feeding a practical diet. Observations during the first year showed t h a t hatchability was most depressed during the earlier part of the egg production year. Analysis of eggs and carcasses from hens fed the low fat diet during the second generation indicated no detectable linoleic, linolenic or arachidonic acid (Table 4).
1017
FATTY ACID D E F I C I E N C Y
Corn diet
"5
5
7-
MonCh8 of Production
FIG. 1. Influence of diet on egg weight (Experiment 1)
5
6 Months of Production
7
FIG. 2. Influence of diet on egg weight (Experiment 2)
Downloaded from http://ps.oxfordjournals.org/ at OhioLINK Offices on May 2, 2015
Btl.l Basil + Safflower oil
1018
L. S. JENSEN AND J. V. SHUTZE
TABLE 4.—Composition of fatty acids in eggs and carcasses of second generation hens {Experiment 2) Fatty acids (%) Diet Low fat
Low fat+5% safflower oil Practical
eggs, first eggs, 6 wks. eggs, 9 wks. carcass, whole eggs, first eggs, 6 wks. eggs, 9 wks. carcass, whole eggs, first eggs, 6 wks. eggs, 9 wks. carcass, whole
14:0!
16:0
16:1
18:0
18:1
18:2
18:3
20:4
0 0 0 0.4 0 0 0 0.4 0 0 0 0.9
26.2 29.1 26.5 23.4 26.7 29.5 26.2 19.2 26.6 28.2 28.2 25.9
2.0 2.0 2.2 9.5 1.3 0.6 1.2 4.2 1.7 2.3 2.3 8.7
8.0 7.2 6.2 3.8 11.2 9.2 11.3 4.3 10.1 7.9 7.6 2.3
63.8 61.6 65.3 62.6 45.0 43.8 44.8 36.7 53.6 52.9 53.7 51.3
0 0 0 0 15.8 17.0 16.9 35.2 7.9 8.8 8.1 10.9
0 0 0 0 0 0 0 0 0 0 0 0
0 0 0 0 0 0 0 0 0 0 0 0
First number indicates chain length, second indicates number of double bonds.
influence on fertility. Because little or no linoleic, linolenic or arachidonic acid was detected in the eggs, and since live, apparently normal chicks were hatched, these fatty acids did not appear to be essential for life in the chicken. Although hatchability of fertile eggs was markedly depressed during the 4th and 5th month of production, the depression was considerably less during the 7th, and almost nonexistent during the 8th month. These results suggest that the increased mortality of embryos may have been associated with egg size, rather than the specific need for essential fatty acids during embryonic development. Average egg weight increased progressively during the experiment for all treatments, even though eggs from hens fed the basal diet were consistently smaller than those from the other treatments. It is possible that essential fatty acids function in the transport of lipids to the ovary. The difference in egg weight was primarily a reflection of the difference in size of the yolk which, in turn may be caused by a slower rate of lipid transport to the ovary. Even though yolk size was considerably lower, time of ovulation apparently was not affected, at least in the first generation birds. A number of reports
using other species of animals have indicated that essential fatty acids are involved in in vivo lipid transport (AaseJorgensen, 1961). In addition to finding no essential fatty acids in egg or carcass samples from birds fed the low fat diet, no trienes (20:3) were detected. An increase in trienes generally has been found in the tissues of animals deficient in essential fatty acids (AaseJorgensen, 1961). The failure to detect these acids may have been caused by sampling lipids from the entire carcass rather than lipids from specific tissues. Machlin and Gordon (1961) detected no essential fatty acids or trienes in the depot fat of chicks fed a low fat diet, but observed significant quantities of these in lipids from heart, liver, cerebrum and testes. The depot fat in hens would have greatly diluted the lipids in other tissues. Until a more detailed analysis of composition of non-depot fat lipids is made, one cannot be sure that the hens used in these studies were completely free of linoleic or arachidonic acid. SUMMARY
Chicks were reared from day of age to maturity on a low fat diet containing a very low level of linoleic acid, and carried
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1
Analysis
FATTY ACID DEFICIENCY
REFERENCES Aase-Jorgensen, E., 1961. Essential fatty acids. Physiol. Rev. 41: 1-51. Bieri, J. G., G. M. Briggs, M. R. Spivey-Fox, C. J. Pollard and L. O. Ortiz, 1956. Essential fatty acids in the chick. 1. Development of fat deficiency. Proc. Soc. Exp. Biol. Med. 9 3 : 237-240. Bieri, J. G., C. J. Pollard and G. M. Briggs, 1957.
Essential fatty acids in the chick. 2. Polyunsaturated fatty acid composition of blood, heart and liver. Arch. Biochem. Biophys. 68: 300-307. Craig, B. M., and N. L. Murty, 1959. Quantitative fatty acid analysis of vegetable oils by gas-liquid chromatography. J. Am. Oil Chem. 36: 549-552. Dam, H., G. Kristensen, G. K. Nielsen, I. Prange and E. Sondergaard, 1956. Influence of varied levels of peanut oil and cholesterol on cholesterol and polyenoic acids in tissues of the chicks. Acta Physiol. Scand. 36: 319-328. Holman, R. T., 1960. The ratio of trienoic: tetraenoic acids in tissue lipids as a measure of essential fatty acid requirement. J. Nutrition, 70: 405-410. Jensen, L. S., J. B. Allred, R. E. Fry and J. McGinnis, 1958. Evidence for an unidentified factor necessary for maximum egg weight in chickens. J. Nutrition, 65: 219-233. Machlin, L. J., and R. S. Gordon, 1961. Effect of dietary fatty acids and cholesterol on growth and fatty acid composition of the chicken. J. Nutrition, 75: 157-164. Murty, N. L., M. C. Williams and R. Reiser, 1960. The non-synthesis of linoleic acid by the laying hen. J. Nutrition, 72: 451-454. Reiser, R., 1950. Fatty acid changes in egg yolk of hens on a fat-free and a cottonseed oil ration. J. Nutrition, 40: 429-440. Reiser, R., 1951. The synthesis and interconversion of polyunsaturated fatty acids by the laying hen. J. Nutrition, 44: 159-175. Shutze, J. V., and L. S. Jensen, 1963. Influence of linoleic acid on egg weight. Poultry Sci. 921-924.
NEWS AND NOTES {Continued from page 1000) Council are: President—H. Ford, Sanderson Frams Inc., Laurel, Miss.; Vice-President—J. A. King, Farm Industries, Decatur, Ala.; Secretary—J. E. Beasley, III, Sweet Sue Poultry Co., Athens, Ala.; Treasurer—J. M. Evans, Jr., Georgia Broilers, Gainesville, Ga. S.E.P.E.A. NOTES At the 15th annual convention of the Southeastern Poultry and Egg Association, the following officers were elected: President—P. Morgan, Guilford College, N. C ; First Vice-President—H. F. McCarty, Magee, Miss.; Second Vice-President— J. P. Wallace, St. Petersburg, Fla.; Treasurer— G. Heitz, Broadway, Va.; Secretary—S. Johnston, Woodstock, Ga.
WPSA NOTES The following are the officers and council members of the World's Poultry Science Association: President—R. C. Blake (Australia); Honorary Past Presidents—H. H. Alp (United States of America), R. Coles (England), A. Ghigi (Italy), W. A. Kock (Denmark), W. D. Termohlen (United States of America) and A. Wiltzer (France); Honorary Past Vice-Presidents—H. Engler (Switzerland), and C. S. Th. Van Gink (Netherlands); Vice-Presidents —M. van Albada (Netherlands), H. R. Bird (United States of America), H. Ebbell (Switzerland), and R. F. Gordon (England); Treasurer and Assistant Secretary—R. G. Jaap (United States of America); Secretary and Assistant Treasurer—I. Macdougall
(Continued on page 1035)
Downloaded from http://ps.oxfordjournals.org/ at OhioLINK Offices on May 2, 2015
through ten months of egg production. A second generation hatched from eggs low in or free of essential fatty acids were reared again to maturity and kept for several months on the low fat diet. Rate of egg production of the first generation was not improved by supplementing the diet with linoleic acid (5% safflower oil), but was slightly improved with the second generation. A marked increased in egg weight was obtained with linoleic acid. Hatchability of fertile eggs, but not fertility, was depressed by the low fat diet. This may have been associated with egg weight, as the depression gradually disappeared during the laying year. No essential fatty acids or trienoic acids were detected in eggs or carcasses from second generation hens, but lipids from specific tissues, not diluted with depot fat, were not analyzed.
1019
A. W. KOTULA, J. E. THOMSON, J. F. NOVOTNY AND E. H. MCNALLY
score for unfrozen thighs, and a 1.3 increase in score for frozen thighs. ACKNOWLEDGMENTS
Essential Fatty Acid Deficiency in the Laying Hen 1 LEO S. JENSEN AND JOHN V. SHUTZE Department of Poultry Science, Washington State University, Pullman (Received for publication February 11, 1963)
AN ESSENTIAL fatty acid deficiency - i * - in the rat has been thoroughly investigated and the symptoms are well defined (Aase-Jorgensen, 1961). Less study has been devoted to such a deficiency in birds. An essential fatty acid deficiency in growing chickens was reported by Bieri et al. (1956) in which chicks fed a fat-free diet grew less after 6 weeks of age than those fed the basal diet supplemented with 4% corn oil. No marked deficiency symptoms were observed, however, even after 20 weeks of depletion. Machlin and Gordon (1961) obtained growth responses to linoleic acid supplements during the first two weeks of the chick's life. The growth depression on the fat-free diet was accentuated by adding saturated fatty acids
1
Scientific Paper No. 2312, Washington Agricultural Experiment Stations, Pullman. Project 1532. Supported in part by funds from Medical and Biological Research by State of Washington Initiative No. 171.
and cholesterol. Fatty acid analysis of heart, testes and cerebrum revealed an increase in triene and decrease in tetraene acids in the deficient birds. Dam et al. (1956) and Bieri et al. (1957) also observed a similar change. An increased ratio of triene to tetraene was orignally observed as a result of an essential fatty acid deficiency in other species, and Holman (1960) has proposed the determination of this ratio in tissues as a criterion for estimating the essential fatty acid requirement in animals. Reiser (1950, 1951) studied an essential fatty acid deficiency in the laying hen and reported that when very low levels of linoleic acid were obtained in eggs, the reproductive performance of the hens apparently was not affected. An earlier conclusion that the hen can synthesize linoleic acid de novo was later reversed by Murty et al. (I960), using more refined techniques. In a previous report from our labora-
Downloaded from http://ps.oxfordjournals.org/ at OhioLINK Offices on May 2, 2015
Appreciation is expressed for the cooperation of the Institute of American Poultry Industries which supplied the chickens; Poultry Nutrition Investigations, Poultry Research Branch, ARS, which prepared diets and maintained the birds; and E. James Koch, ARS, U. S. Department of Agriculture, for advice and assistance in analyzing the data.
REFERENCES Brant, A. W., and G. F. Stewart, 1950. Bone darkening in frozen poultry. Food Tech. 4:168-174. Duncan, D. B., 1955. Multiple range and multiple F tests. Biometrics, 11: 1-7. Ellis, C , and J. G. Woodroof, 1959. Prevention of bone darkening. Food Tech. 13: 533-538. Essary, E. 0., and E. L. Wisman, 1962. Personal communication. Koonz, C. H., 1946. Poultry and Eggs. Refrig. Eng. Data Book 59. Woodroof, J. G., and E. Shelor, 1948. Prevention of bone darkening in frozen-packed chickens. Food Industries, 20: 48-52.
1015
FATTY ACID DEFICIENCY
tory, corn oil was shown to increase egg weight of birds fed a semi-purified diet (Jensen et al., 1958). Hatchability was also shown to be adversely affected after some time on the semi-purified diet unless corn was included. Linoleic acid was found to be the major factor in corn oil responsible for the egg weight increase (Shutze and Jensen, 1963). The present experiments were undertaken to study further an essential fatty acid deficiency in the laying hen.
Single Comb White Leghorn chicks were reared on three different diets from day of age to maturity and were then fed similar diets during approximately ten months of egg production. The chicks were brooded in battery brooders with raised wire floors and were then transferred to wire floored colony cages with about 16 birds per pen. At 20 weeks of age the birds were moved to individual hen cages. Feed and water were supplied ad libitum. In order to obtain fertile eggs, the hens were artificially inseminated with semen from males maintained on a practical breeder diet. Chicks hatched from eggs laid by the hens fed the three diets were reared to maturity and kept for 39 weeks in production. Composition of the basal low fat and practical laying diet is shown in Table 1. Rearing diets were similar, except that level of protein was higher for the first 8 weeks and calcium carbonate was completely removed. No linoleic acid was detected in the low fat basal diet by gasliquid chromatography, whereas the low fat diet supplemented with safflower oil and the practical diet contained considerable linoleic acid. All unbroken eggs laid were weighed at two week intervals. Samples of eggs from each treatment were incubated to deter-
Layer diets Ingredients Low Fat Practical Corn, yellow Cerelose Defatted fish meal1 Soybean meal, dehulled Fish meal, herring Dehydrated alfalfa meal Dried brewers' yeast Limestone Dicalcium phosphate NaCl, iodized Manganese sulfate KC1 Trace mineral mixture Vitamin mixture
% 62.80 30.00
— — —
3.00 3.20
—
0.40
—
0.40 0.05 2 1.003
%
74.75
— —
8.00 5.00 3.00 2.00 3.25 3.00 0.50 0.02
— —
0.50"
1 Obtained from Viobin Corporation, Monticello, Illinois. 2 Supplied the following p.p.m.: Mn, 50.0; Fe, 50.0; Cu, 5.0; Co, 0.5; I, 1.5; Zn, 50.0. 3 Supplied the following per kg. of diet: vitamin A, 8,811 I.U.; vitamin D 3 , 881 I.U.; menadione sodium bisulfite, 6.6. mg.; vitamin E, 11.0 I.U.; riboflavin, 4.4. mg.; pyridoxine, 4.4 mg.; Ca-pantothenate, 13.9 mg.; niacin, 22.0 mg.; folic acid, 0.44 mg.; B12, 2.2 meg.; ethoxyquin, 249.; bacitracin, 11 mg. 1 Supplied the following per kg. of diet: vitamin A, 6,6101.U.; vitamin D 3 , 7441.U.; riboflavin, 4.4. mg.; vitamin E, 22.0 I.U.; Pn2, 9.9 meg.; ethoxyquin, 249 mg.; bacitracin, 11.0 mg. Note: The safflower diet consisted of adding 5% safflower oil to the low fat diet at the expense of cerelose for the first experiment. During the second experiment, 11.2% of the cerelose was left out and only 5% safflower oil was added in order to make these two diets isocaloric.
mine the effect on fertility and hatchability. Fatty acids were determined by a gas-liquid chromatography method as follows: the dried feed samples and lyophilized egg samples were extracted for 12 hours in a Soxhlet with a 2:1 mixture of chloroform and methanol. The carcasses were ground through a meat grinder and thoroughly mixed before an aliquot was taken for lyophilization and extraction. Methyl esters were formed by interesterification, using sodium methoxide as a catalyst (Craig and Murty, 1959). A Beckman GC2A model instrument with a thermol conductivity detector was used for analysis of the methyl esters. A | in.
Downloaded from http://ps.oxfordjournals.org/ at OhioLINK Offices on May 2, 2015
EXPERIMENTAL
TABLE 1.—Composition of diets
1016
L. S. J E N S E N AND J. V. SHUTZE
TABLE 2.—EJfect of an essential fatty acid deficiency on egg production % Average egg production Dlet
First generation 1
Second generation 2
Low fat basal Low fat basal + 5 % safflower oil Practical
44.6
29.1
45.3 56.5
38.0 56.7
1 1
Average for 42 week period. Average for 39 week period.
RESULTS
Although the chicks fed the basal diet supplemented with safflower oil grew faster t h a n the chicks fed the unsupplemented basal diet, both groups reached sexual maturity at about the same age. Chicks fed the practical diet grew at about the same rate as those on the unsupplemented basal diet. Similar observations were made with the second generation chicks. There was no difference in r a t e of lay between pullets fed the basal diet and the diet supplemented with safflower oil during the first experiment (Table 2). However, birds fed the practical diet laid at a faster rate. In the second experiment (involving off-spring from the initial birds used in the study), slightly lower egg production was obtained with the unsupplemented basal diet. Again, birds fed the practical diet laid a t a more rapid rate, suggesting t h a t factors other t h a n essential fatty acids were influencing egg production. Egg weight was markedly influenced by the diet. Average egg weight of birds fed the diet supplemented with safflower oil was consistently 2-6 grams heavier t h a n
DISCUSSION Essential fatty acid deficiency in the laying hen resulted in a marked reduction in egg weight, a depression in hatchability of fertile eggs, but had little effect on rate of egg production and apparently no TABLE 3.—Effect of an essential fatly acid deficiency in hens on fertility and hatchability Diet
Month of production
First generation Low fat basal Low fat basal plus safflower oil Practical Low fat basal Low fat basal plus safflower oil Practical Low fat basal Low fat basal plus safflower oil Practical Low fat basal Low fat basal plus safflower oil Practical Second generation Low fat basal Low fat basal plus safflower oil Practical
No. eggs set
Fertility
% Hatch of fertile eggs
%
4
111
73.6
38.3
4 4
178 215
49.4 46.5
62.5 72.0
S
111
72.5
33.7
5 5
137 125
84.7 87.2
68.0 86.3
7
138
79.0
56.8
7 7
212 180
83.0 73.3
68.2 84.1
8
144
66.6
79.3
8 8
195 281
79.0 80.0
81.3 78.7
6-7
318
74.2
61.0
6-7 6-7
508 744
73.6 88.3
71.1 84.8
•
Downloaded from http://ps.oxfordjournals.org/ at OhioLINK Offices on May 2, 2015
X12 ft. copper column was packed with 2 5 % diethylene glycol succinate absorbed on 40-60 mesh acid washed chromosorb W and helium gas was used as a carrier. Column temperature was maintained at 235° C.
that of birds fed the unsupplemented basal diet (Figs. 1 and 2). Eggs from hens fed the practical diet were consistently heavier than both the groups fed the purified diets, indicating t h a t factors other than essential fatty acids were also influencing egg weight. No consistent effect of diet on fertility was observed (Table 3), but hatchability of fertile eggs was increased by supplementing the purified diet with safflower oil or by feeding a practical diet. Observations during the first year showed t h a t hatchability was most depressed during the earlier part of the egg production year. Analysis of eggs and carcasses from hens fed the low fat diet during the second generation indicated no detectable linoleic, linolenic or arachidonic acid (Table 4).
1017
FATTY ACID D E F I C I E N C Y
Corn diet
"5
5
7-
MonCh8 of Production
FIG. 1. Influence of diet on egg weight (Experiment 1)
5
6 Months of Production
7
FIG. 2. Influence of diet on egg weight (Experiment 2)
Downloaded from http://ps.oxfordjournals.org/ at OhioLINK Offices on May 2, 2015
Btl.l Basil + Safflower oil
1018
L. S. JENSEN AND J. V. SHUTZE
TABLE 4.—Composition of fatty acids in eggs and carcasses of second generation hens {Experiment 2) Fatty acids (%) Diet Low fat
Low fat+5% safflower oil Practical
eggs, first eggs, 6 wks. eggs, 9 wks. carcass, whole eggs, first eggs, 6 wks. eggs, 9 wks. carcass, whole eggs, first eggs, 6 wks. eggs, 9 wks. carcass, whole
14:0!
16:0
16:1
18:0
18:1
18:2
18:3
20:4
0 0 0 0.4 0 0 0 0.4 0 0 0 0.9
26.2 29.1 26.5 23.4 26.7 29.5 26.2 19.2 26.6 28.2 28.2 25.9
2.0 2.0 2.2 9.5 1.3 0.6 1.2 4.2 1.7 2.3 2.3 8.7
8.0 7.2 6.2 3.8 11.2 9.2 11.3 4.3 10.1 7.9 7.6 2.3
63.8 61.6 65.3 62.6 45.0 43.8 44.8 36.7 53.6 52.9 53.7 51.3
0 0 0 0 15.8 17.0 16.9 35.2 7.9 8.8 8.1 10.9
0 0 0 0 0 0 0 0 0 0 0 0
0 0 0 0 0 0 0 0 0 0 0 0
First number indicates chain length, second indicates number of double bonds.
influence on fertility. Because little or no linoleic, linolenic or arachidonic acid was detected in the eggs, and since live, apparently normal chicks were hatched, these fatty acids did not appear to be essential for life in the chicken. Although hatchability of fertile eggs was markedly depressed during the 4th and 5th month of production, the depression was considerably less during the 7th, and almost nonexistent during the 8th month. These results suggest that the increased mortality of embryos may have been associated with egg size, rather than the specific need for essential fatty acids during embryonic development. Average egg weight increased progressively during the experiment for all treatments, even though eggs from hens fed the basal diet were consistently smaller than those from the other treatments. It is possible that essential fatty acids function in the transport of lipids to the ovary. The difference in egg weight was primarily a reflection of the difference in size of the yolk which, in turn may be caused by a slower rate of lipid transport to the ovary. Even though yolk size was considerably lower, time of ovulation apparently was not affected, at least in the first generation birds. A number of reports
using other species of animals have indicated that essential fatty acids are involved in in vivo lipid transport (AaseJorgensen, 1961). In addition to finding no essential fatty acids in egg or carcass samples from birds fed the low fat diet, no trienes (20:3) were detected. An increase in trienes generally has been found in the tissues of animals deficient in essential fatty acids (AaseJorgensen, 1961). The failure to detect these acids may have been caused by sampling lipids from the entire carcass rather than lipids from specific tissues. Machlin and Gordon (1961) detected no essential fatty acids or trienes in the depot fat of chicks fed a low fat diet, but observed significant quantities of these in lipids from heart, liver, cerebrum and testes. The depot fat in hens would have greatly diluted the lipids in other tissues. Until a more detailed analysis of composition of non-depot fat lipids is made, one cannot be sure that the hens used in these studies were completely free of linoleic or arachidonic acid. SUMMARY
Chicks were reared from day of age to maturity on a low fat diet containing a very low level of linoleic acid, and carried
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Analysis
FATTY ACID DEFICIENCY
REFERENCES Aase-Jorgensen, E., 1961. Essential fatty acids. Physiol. Rev. 41: 1-51. Bieri, J. G., G. M. Briggs, M. R. Spivey-Fox, C. J. Pollard and L. O. Ortiz, 1956. Essential fatty acids in the chick. 1. Development of fat deficiency. Proc. Soc. Exp. Biol. Med. 9 3 : 237-240. Bieri, J. G., C. J. Pollard and G. M. Briggs, 1957.
Essential fatty acids in the chick. 2. Polyunsaturated fatty acid composition of blood, heart and liver. Arch. Biochem. Biophys. 68: 300-307. Craig, B. M., and N. L. Murty, 1959. Quantitative fatty acid analysis of vegetable oils by gas-liquid chromatography. J. Am. Oil Chem. 36: 549-552. Dam, H., G. Kristensen, G. K. Nielsen, I. Prange and E. Sondergaard, 1956. Influence of varied levels of peanut oil and cholesterol on cholesterol and polyenoic acids in tissues of the chicks. Acta Physiol. Scand. 36: 319-328. Holman, R. T., 1960. The ratio of trienoic: tetraenoic acids in tissue lipids as a measure of essential fatty acid requirement. J. Nutrition, 70: 405-410. Jensen, L. S., J. B. Allred, R. E. Fry and J. McGinnis, 1958. Evidence for an unidentified factor necessary for maximum egg weight in chickens. J. Nutrition, 65: 219-233. Machlin, L. J., and R. S. Gordon, 1961. Effect of dietary fatty acids and cholesterol on growth and fatty acid composition of the chicken. J. Nutrition, 75: 157-164. Murty, N. L., M. C. Williams and R. Reiser, 1960. The non-synthesis of linoleic acid by the laying hen. J. Nutrition, 72: 451-454. Reiser, R., 1950. Fatty acid changes in egg yolk of hens on a fat-free and a cottonseed oil ration. J. Nutrition, 40: 429-440. Reiser, R., 1951. The synthesis and interconversion of polyunsaturated fatty acids by the laying hen. J. Nutrition, 44: 159-175. Shutze, J. V., and L. S. Jensen, 1963. Influence of linoleic acid on egg weight. Poultry Sci. 921-924.
NEWS AND NOTES {Continued from page 1000) Council are: President—H. Ford, Sanderson Frams Inc., Laurel, Miss.; Vice-President—J. A. King, Farm Industries, Decatur, Ala.; Secretary—J. E. Beasley, III, Sweet Sue Poultry Co., Athens, Ala.; Treasurer—J. M. Evans, Jr., Georgia Broilers, Gainesville, Ga. S.E.P.E.A. NOTES At the 15th annual convention of the Southeastern Poultry and Egg Association, the following officers were elected: President—P. Morgan, Guilford College, N. C ; First Vice-President—H. F. McCarty, Magee, Miss.; Second Vice-President— J. P. Wallace, St. Petersburg, Fla.; Treasurer— G. Heitz, Broadway, Va.; Secretary—S. Johnston, Woodstock, Ga.
WPSA NOTES The following are the officers and council members of the World's Poultry Science Association: President—R. C. Blake (Australia); Honorary Past Presidents—H. H. Alp (United States of America), R. Coles (England), A. Ghigi (Italy), W. A. Kock (Denmark), W. D. Termohlen (United States of America) and A. Wiltzer (France); Honorary Past Vice-Presidents—H. Engler (Switzerland), and C. S. Th. Van Gink (Netherlands); Vice-Presidents —M. van Albada (Netherlands), H. R. Bird (United States of America), H. Ebbell (Switzerland), and R. F. Gordon (England); Treasurer and Assistant Secretary—R. G. Jaap (United States of America); Secretary and Assistant Treasurer—I. Macdougall
(Continued on page 1035)
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through ten months of egg production. A second generation hatched from eggs low in or free of essential fatty acids were reared again to maturity and kept for several months on the low fat diet. Rate of egg production of the first generation was not improved by supplementing the diet with linoleic acid (5% safflower oil), but was slightly improved with the second generation. A marked increased in egg weight was obtained with linoleic acid. Hatchability of fertile eggs, but not fertility, was depressed by the low fat diet. This may have been associated with egg weight, as the depression gradually disappeared during the laying year. No essential fatty acids or trienoic acids were detected in eggs or carcasses from second generation hens, but lipids from specific tissues, not diluted with depot fat, were not analyzed.
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