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Calcium and Phosphorus in the Diet of Coturnix Quail1
686
P. L. COX, R. D . HOLLIFIELD AND J. P. HEOTIS
80° C. and increase the temperature to 85°C. in order to avoid boiling over and loss of compound. During the method development there was daily variation in reagent blanks and control values. With this variation the standards and recoveries were acceptable when corrected for the average reagent blank or control value obtained simultaneously with the particular assay. To allow for possible daily variation in control values when assaying samples or in reagent blanks when assaying standards, control samples or reagent blanks should be included and the results calculated based on the control or reagent blank value obtained for that particular assay. The use of quinine sulfate as a primary standard for adjusting the sensitivity of the instrument each day eliminates the need of including a complete set of standards each day. Concentrations can be determined from a previously prepared standard curve, provided the same arbitrary fluorescence value is used in standardizing the instrument with quinine sulfate. The procedure is rather time consuming but the chloroform extracts and 80% methanol solutions containing buquinolate are stable overnight if stored in the refrigerator.
SUMMARY
A spectrophotofluorometric method developed for the determination of buquinolate in poultry tissue and eggs is sensitive to 0.1 p.p.m. Tissues were assayed from chickens following administration of the compound to broilers for 9-10 weeks in feed at the 75 and 100 gram per ton level, and to layers at 100 grams per ton of feed for 18 months. Levels of less than 0.1 to 0.39 p.p.m. were detected in liver, kidney, fat and skin at zero withdrawal, while muscle contained less than 0.1 p.p.m. at zero withdrawal. Tissues collected following 1 or 2 days withdrawal from buquinolate contained less than 0.1 p.p.m. with the exception of an occasional sample that contained levels of the order of 0.1 p.p.m. Eggs collected from layers following the administration of buquinolate at 100 grams per ton of feed for 18 months contained from 0.11 to 0.23 p.p.m. of buquinolate. REFERENCES Engle, A. T., R. P. Humphrey and C. Johnson, 1966. Buquinolate, a new broad spectrum coccidiostat. Poultry Sci. 4S: 1082. Herrett, R. J., G. M. Klein, C. W. Williams, and J. P. Heotis, 1966. The distribution of C" in the chick following the administration of Buquinolate-3-C". Poultry Sci. 45: 1092.
Calcium and Phosphorus in the Diet of Coturnix Quail1 B.
F.
MILLER
Department of Poultry Science, Colorado Agricultural Experiment Station, Fort Collins, Colo. 80521 (Received for publication October 7, 1966)
INTRODUCTION
T
HE Coturnix quail (Coturnlx coturnix japonica) provides a relatively new and increasingly used tool for re1
Published with the approval of the Director of the Colorado Agricultural Experiment Station as S. S. paper 1144, journal series.
search. Its characteristics and requirements are not well explored. Johnson (1960) in a review of the usefulness of the Coturnix emphasized that the quail has many advantages over other laboratory animals including resistance to disease and relatively small maintenance cost. This animal is easily handled, reproduces in captivity readily
CA AND P IN QUAIL DIETS
687
and requires little space and feed. Howes and Cassidy (1964b) found similar results and Frey (1961) have outlined a procedure when the study of phosphorus was confor obtaining and caring for the Coturnix tinued to eight weeks of age. Feed intake quail; while Wilson et al. (1961) have in- and body weight at eight weeks of age and vestigated reproductive capacities and ob- tibial bone ash at four weeks of age were tained data concerning fertility, hatchabili- improved in chicks receiving supplementy, growth rate and egg production. tary inorganic phosphorus. The National Research Council (1966) Turkey poults (Temperton and Cassirecommended one percent calcium and 0.6 dy, 1964c) fed an all-mash diet containing percent phosphorus in the diet of starting no feedstuffs of animal origin and with no and growing chicks. For starting poults the phosphorus supplement developed severe recommendations are increased to 1.2 per- rickets characterized by bowing of the leg cent calcium and 0.8 percent phosphorus. bones and beading of the ribs. Affected One percent phosphorus is recommended poults recovered and clinical symptoms disfor starting and growing pheasants, but no appeared as a result of adding sufficient sorecommendation is given in regard to dium phosphate to provide 0.9 percent young quail for either calcium or phospho- phosphorus in the diet. rus. For quail breeders, 2.3 percent calcium Temperton et al. (1965a) showed that and 1.0 percent phosphorus are recom- with light weight hybrid pullets, diets conmended. taining 0.45 percent total phosphorus and Sunde and Bird (1956) working with 0.14 percent available phosphorus during pheasant chicks reported that feeding a the first eight weeks did not result in any diet containing 0.66 percent phosphorus significant depression of body weight, feed (analytical analysis) resulted in high mor- consumption, feed efficiency or livability tality. The pheasant chicks after two weeks when compared to birds receiving diets on this diet showed a distinct bending of supplemented to the N.R.C. recommended the tarsometatarsus near the hock joint levels with inorganic phosphorus. Temperand at the proximal end of the tibiotarsus. ton et al. (1965b) further observed no Bone ash values were reduced from a nor- marked effects on total bone ash, calcium mal of 52 to 38 percent when the basal diet or phosphorus content of toes from feeding was fed. Additions of 0.3 to 0.8 percent of a diet containing 0.46 percent total phosphosphorus from dicalcium phosphate re- phorus and 0.14 percent available phosphosulted in normal growth, normal bone for- rus from one-day old to eight weeks of age. The amount of phosphorus in the toes at mation and calcification. Temperton and Cassidy (1964a) have 18 weeks of age was approximately oneshown that pullet chicks can grow normally half the amount of calcium in all treatup to 26 days of age receiving a vegetable ments. protein diet containing 0.48 percent total Plant source phosphorus (Vandepopuphosphorus and 0.23 percent non-phytin liere et al., 1961) was readily available to phosphorus. Balance studies indicated support growth when fed to broiler chicks these chicks retained approximately 60 per- at a calcium to phosphorus ratio of 1:1; cent of the phytin phosphorus ingested. however, it did not allow for increased The percent bone ash, calcium and phos- bone ash as did dicalcium phosphorus. phorus concentrations and calcium to phos- Chicks which had bone ash values of 28 phorus ratio of tibiae reflected normal bone percent did not exhibit symptoms of rickets formation. These same workers, Temperton when fed a diet containing 0.28 percent
688
B. F. MILLER
plant phosphorus and a calcium to phosphorus ratio of 1:1. A phosphorus level of 0.72 percent from dicalcium phosphate with a Ca:P ratio of 1:1 to 1.5:1 produced maximum growth in battery trials. The criterion, growth, appeared to be sufficient to determine the phosphorus adequacy of a diet when the level approached the requirement. It was essential to consider both calcium and phosphorus levels and the calcium to phosphorus ratio when determining phosphorus availability, irrespective of the phosphorus source. Nelson et al. (1964) reported that Coturnix quail breeders produced better when they received 2.5 and 3.0 percent calcium as compared to birds that received 1.0, 1.5 and 2.0 percent calcium in their diet. Little difference in egg production was found between groups receiving 0.6 and 0.8 percent phosphorus. There was little difference in fertility between treatments, but hatchability was variable with higher hatchability on the 2.5 and 3.0 percent calcium diets and on 0.8 percent phosphorus. Very little information appears in the literature about the nutritive requirements of the Coturnix quail. In view of the increasing acceptance of Coturnix as a laboratory animal and the apparent lack of information of their nutritional requirements, this study was initiated. In addition, it was hoped to add to the limited knowledge of the phosphorus and calcium requirements of Coturnix quail as well as add to the overall understanding of the role of phosphorus in the diet. PROCEDURE
Twelve pens of one-day old Coturnix quail with 15 chicks per pen were utilized in the first trial. The chicks were randomly distributed among the pens. The different treatments were randomly arranged within a modified Jamesway battery brooder. Modification consisted of covering
the floor and sides of each pen with onequarter inch mesh screen to prevent the chicks from escaping. The temperature of the brooders was adjusted to 37°C. during the first week of brooding. Room temperature was held at approximately 23°C. The brooder temperature was decreased 3°C. each week thereafter until room temperature was reached. The birds received 14 hours of light from fluorescent fixtures placed overhead in the room. The room was air conditioned and force ventilated. During the first two weeks, quail feeders were utilized to feed the chicks. One quart glass chick waterers were utilized for watering during this time. The chick waterers were modified by filling the lower half of the watering area with plastic cement to prevent the quail chicks from getting into the deep water and drowning. Feed was placed on paper towels during the first three days to encourage eating. After the first two weeks the standard battery feeders and waterers were utilized for the growing quail. Feed consumption was measured weekly. Wastage of feed during the first two weeks was excessive in some lots making these data unreliable. Feed wastage during the remainder of the experiment was slight and fairly uniform from one experimental group to another. The average body weights for each group were determined weekly by weighing the group and dividing by the number of birds within the group. Mortality was recorded daily; however, any birds that died during the first week of the experiment were replaced with comparable aged birds receiving the basal (control) diet. The basal diet (Table 1) had an analyzed calcium level of 0.44 percent and a phosphorus level of 0.59 percent. Ground limestone and dicalcium phosphate were the only ingredients added to increase the level of calcium and phosphorus for the various experimental diets. The different levels of calcium and phosphorus studied
689
CA AND P IN QUAIL DIETS
are shown in Table 3. A random sample of 10 birds from each group was individually scored for proper feathering with 10 indicating a perfect covering of feathers. At six weeks of age, when the experiment was terminated, one male and one female were sacrificed from each group, and the tibiae were analyzed for percent ash of moisture and fat-free bone. In the second trial, the same procedure was followed; however, each experimental group was replicated with 10 birds per experimental group. During the sixth week of the experiment all surviving birds were individually feather scored as before. Comparisons of average body weight, feed efficiency, percent bone ash, livability, and feather scores were made among the different treatments. Samples of the feed were analyzed for phosphorus using the molybovandophosphoric acid procedure with wet digestion in perchloric acid (Barton, 1948). Calcium was determined on the atomic absorption TABLE 1.—Composition of basal diet utilized to study phosphorus requirement of Coturnix quail Ingredient
Percent
Corn, ground yellow Soybean meal (50% protein) Alfalfa meal (20% protein) Added per 45.4 kg. of diet: Vitamin A Vitamin D 3 Vitamin E Vitamin K Thiamine Riboflavin Pyridoxine Pantothenic acid Folacin Niacin Biotin Vitamin B12 Choline Salt and trace mineral mix: NaCl, iodized Mg Mn Zn Fe Cu Mo Methionine
41. 5 51. 0 5. 0 400,000 100,000 1,000 50 100 200 100 400 40 3,000 4 0.5 20 136 200 50 50 25 3 2 50
I.U. I.U. I.U. mg. mg. mg. mg. mg. mg. mg. mg. mg. gm. gm. p.p.m. p.p.m. p.p.m. p.p.m. p.p.m. p.p.m. gm.
TABLE 2.—Comparison of mean weekly body weights and mean weekly body weights as compared to the weights reported by Wilson et al. {1961) Age of birds (weeks) 1 Mean body weight (gm.) (Trials 1 and 2) Mean body weights 1 (gm.) 1
,2
3
22 40 65 38 64 84
4
5
84 100
94 111
Wilson et al. (1961).
spectrophotometer after digestion.
the above wet
RESULTS AND DISCUSSION
The basal diet containing 0.44 percent calcium and 0.59 percent phosphorus on an analytical basis resulted in satisfactory growth and development (Table 3). Body weights and growth rate of the birds that received the basal diet were above the average for all treatment groups. Percent bone ash was not depressed to any significant extent by the low levels of calcium and phosphorus in the diet. Feather scores of these birds were the highest in all cases. The birds that received the basal diet showed no ill effects from consuming this diet. Livability was not affected by the diet. Feathering was significantly enhanced by this treatment over some of the other diets utilized in the study. Statistical analysis of mean body weights indicated there were no significant differences among treatments. Average body weights and Ca:P ratios of the diets used are presented in Table 3. The greatest body weight was found where the birds received 0.70 percent Ca and 0.58 percent P in the diet. Mean body weights (Table 2) were compared by weeks to those reported by Wilson et al. (1961). The birds in this experiment were approximately one week behind in their weights. These birds were slow in getting started as indicated by their one-week old weights. They reached sexual maturity at about five to six weeks of age,
690
B. F. MILLER
TABLE 3.—Effects of dietary calcium and phosphorus levels upon quail body weights and percent bone ash at six weeks of age1
Diet
Calcium2 percent
11 (Basal) 21 14 18 19 12 20 15 13 22 16 17
0.44 1.12 0.90 1.08 1.29 0.70 1.14 1.41 1.08 2.30 1.58 2.18
Phosphorus percent2
Ca:P ratio
Average body wt.* (grams)
Percent bone ash
0.59 1.18 0.83 1.02 1.14 0.58 0.86 0.76 0.58 1.14 0.74 0.75
0.7:1 1.0:1 1.1:1 1.1:1 1.1:1 1.2:1 1.3:1 1.8:1 1.9:1 2.0:1 2.1:1 2.9:1
101 98 101 99 96 103 98 98 97 101 98 100
28 31 31 32 31 28 31 29 30 30 30 31
1 Combined body weights of birds in Trials 1 and 2. ! Calcium and phosphorus level determined by chemical analysis of diet 3 No significant difference between body weights at the 5 percent level.
similar to results reported by Wilson et al. (1961). The birds that were fed diet 12 were heaviest, averaging 103.2 gm. (Table 3) on a diet containing 0.7 percent calcium and 0.58 percent phosphorus. Their feathering was comparable to the controls in trial 2 (Table 5) and somewhat depressed in trial 1 (Table 4). Tables 4 and 5 present the analysis of variance of the mean feather scores during trials 1 and 2, respectively. When the multiple range test (Duncan, 1955) was applied to the treatment means in both exper-
iments, the basal diet (0.7:1 Ca:P ratio) and diet 14 (1.1:1 Ca:P ratio) consistently ranked in the highest group although they are not significantly better than all treatments. Birds receiving diets 13, 15, 16, 17, 20 and 22 (1.3:1 to 2.9:1 Ca:P ratios) were significantly not as well feathered as those birds receiving the basal diet or diet 14 (0.90% Ca and 0.83% P ) . Birds receiving diet 22 (2.3 Ca and 1.1 P) were consistently low in feather score, while the other groups showed considerable variation. Analysis of the feather scores verified the visual observations made during the experiment. Figure 1 provides a comparison of the typical feathering of a bird that received the basal diet and one that received diet 20. Analysis of the data on bone ash indicated there was no significance between treatments (Table 3). The females had a slightly higher bone ash content than the males. The average bone ash content of fat-free tibia was 29.8 percent with a range of 25.8 to 37.0 percent. No significant differences were found in feed efficiency between the various levels of calcium and phosphorus. None of the birds died after the first week of age, except accidental deaths during handling. All diets
TABLE 4.—Analysis of variance of mean feather scores in Trial 1 Sources
D.F.
S.S.
M.S.
F.
129.9 19.0 30.S
4.26 .62
Treatment Period Error
11 1 11
1,429 19 346
Total
23
1,894
Duncan's test of mean feather scores, Trial 1 } Treatment 11 14 19 16 12 (Basal) Avg. 9.4 9.3 9.2 8.2 8.1
Sig.
* N.S.
17
21
20
22
18
IS
13
8.0
8.0
7.9
7.8
7.S
7.3
6.9
* Significant at the five percent level. N.S. Not significant. Any groups underlined by the same line are not statistically different from each other at the P<0.05 level. 1
CA AND P IN QUAIL DIETS
691
TABLE 5.—Analysis of variance of individual bird feather scores at six weeks of age in Trial 2 Sources
D.F.
s.s.
M.S.
Treatment Error
11 203
151
13.73 .96
Total
214
345
19-4
Duncan's test of mean feather scores, Trial 2.1 Treatment 11 14 12 18 21 (Basal) Avg. 9.8 9.4 9.3 9.0 8.9
Sig. 14.30
IS
10
19
13
22
20
17
8.3
8.3
8.3
8.2
7.9
7.1
6.8
** Significant at the one percent level. Any groups underlined by the same line are not statistically different from each other at the P<0.01 level. 1
were capable of sustaining the growing quail to maturity (six weeks of age). SUMMARY One-day old Coturnix quail chicks were reared to maturity (six weeks of age) on
4 f \
Bfc M& '^..
FIG. 1. The bird on the left received the basal diet containing 0.44 percent calcium and 0.59 percent phosphorus while the bird on the right received a similar diet containing 1.14 percent calcium and 0.86 percent phosphorus (diet 20).
an all-vegetable diet containing various calculated levels of calcium and phosphorus. The basal diet contained 0.44 percent calcium and 0.59 percent phosphorus on an analytical basis. Calcium and phosphorus content of the different diets was modified by adding ground limestone and dicalcium phosphate to the basal for the different levels and ratios of calcium and phosphorus studied. The basal diet containing 0.44 percent calcium and 0.59 percent phosphorus resulted in satisfactory gains in body weight. There was no evident impairment of bone ash in birds receiving this diet. Birds that received diets with 0.7:1 and 1.1:1 calcium to phosphorus ratios in general showed significantly better feathering than birds fed diets with 2:1 or higher Ca:P ratios. This was especially true when levels of calcium and phosphorus reached two percent and one percent, respectively. There was no significant difference in feed efficiency or livability between the various diets. Coturnix quail chicks are capable of utilizing an all-vegetable diet that is low in calcium and phosphorus providing it is adequately supplied with vitamins and other minerals. The level of calcium and phosphorus in
692
B. F. MILLER
the diet and the Ca:P ratio had a significant effect on the feathering of growing quail. As the calcium and phosphorus content of the diet increased, the relative feathering of the birds decreased. A spreading of the Ca:P ratio also caused a similar depression in feathering. ACKNOWLEDGMENTS
This investigation was supported in part by a grant-in-aid from the Faculty Improvement Committee, Colorado State University, to whom the author is grateful. Appreciation is expressed to The Ray Ewing Company Division of HoffmannLaRoche, Inc., Pasadena, California, for supplying the vitamin supplements for this study. The assistance of Dr. Eldon W. Kienholz and Dr. Cheryl F. Nockels during the conduct of this study and preparation of the manuscript has been very helpful. REFERENCES Barton, C. J., 1948. Photometric analysis of phosphoric rock. Anal. Chem. 20: 1068-1073. Duncan, D. B., 19SS. Multiple range and multiple F tests. Biometrics, 11: 1^2. Howes, J. R., and W. D. Frey, 1961. Coturnix quail for avian research. Feedstuffs, 33, (21) May 27, 1961. Johnson, E. L., 1960. A new research tool . . . Coturnix quail. Minnesota Farm and Home Science, 18(1): 1. National Research Council of America, 1966. Nutrient requirements of poultry. National Acad-
emy of Sciences-National Research Council, Washington, D.C. Publication 1345. Nelson, F. E., J. K. Lauber and L. Mirosh, 1964. Calcium and phosphorus requirement for the breeding Coturnix quail. Poultry Sci. 43: 1346. Sunde, M. L., and H. R. Bird, 1956. A critical need of phosphorus for the young pheasant. Poultry Sci. 35: 424-^30. Temperton, H., and J. Cassidy, 1964a. Phosphorus requirements of poultry. I. The utilization of phytin phosphorus by the chicks as indicated by balance experiments. British Poultry Sci. 5: 75-80. Temperton, H., and J. Cassidy, 1964b. II. The utilization of phytin phosphorus by the chick for growth and bone formation. British Poultry Sci. S: 81-86. Temperton, H., and J. Cassidy, 1964c. III. The effect of feeding a vegetable-type diet without supplementary phosphorus to turkey poults. British Poultry Sci. 5: 87-88. Temperton, H., F. J. Dudley and G. J. Pickering, 1965a. Phosphorus requirements of poultry. IV. The effects on growing pullets of feeding diets containing no animal protein or supplementary phosphorus. British Poultry Sci. 6: 125-133. Temperton, H., F. J. Dudley and G. J. Pickering, 1965b. The phosphorus requirements of growing pullets between 8 and 18 weeks of age. British Poultry Sci. 6: 143-152. Vandepopuliere, J. M., C. B. Ammermon and R. H. Harms, 1961. The relationship of calciumphosphorus ratios on the utilization of plant and inorganic phosphorus by the chick. Poultry Sci. 40: 951-957. Wilson, W. O., U. K. Abbott and H. Abplanalp, 1961. Evaluation of Coturnix (Japanese quail) as pilot animal for poultry. Poultry Sci. 40: 651-657.
NEWS AND NOTES (continued from page 650) sociation are: President—E. E. Penionzhkevich (U.S.S.R.); Hon. Past Presidents—H. H. Alp (U.S.A.), R. C. Blake (Australia), R. Coles (England), A. Ghigi (Italy), W. D. Termohlen (U.S.A.), A. Wiltzer (France); Vice-Presidents—Z. BenAdam (Israel), W. P. Blount (England), R. Pero (France); Hon. Past Vice-Presidents—H. Engler (Switzerland), C. S. Th. Van Gink (Netherlands); W. P. S. A. NOTES Treasurer and Assistant Secretary—R. G. Jaap The officers of the World's Poultry Science As(U.S.A.); Secretary and Assistant Treasurer—I. (continued < n page 754)
stories of how several companies who are now leaders in the production of further processed poultry and egg products got that way. Further information can be obtained by writing to Dr. George Mountney, Department of Poultry Science, Ohio State University, 674 West Lane Avenue, Columbus, Ohio 43210.
P. L. COX, R. D . HOLLIFIELD AND J. P. HEOTIS
80° C. and increase the temperature to 85°C. in order to avoid boiling over and loss of compound. During the method development there was daily variation in reagent blanks and control values. With this variation the standards and recoveries were acceptable when corrected for the average reagent blank or control value obtained simultaneously with the particular assay. To allow for possible daily variation in control values when assaying samples or in reagent blanks when assaying standards, control samples or reagent blanks should be included and the results calculated based on the control or reagent blank value obtained for that particular assay. The use of quinine sulfate as a primary standard for adjusting the sensitivity of the instrument each day eliminates the need of including a complete set of standards each day. Concentrations can be determined from a previously prepared standard curve, provided the same arbitrary fluorescence value is used in standardizing the instrument with quinine sulfate. The procedure is rather time consuming but the chloroform extracts and 80% methanol solutions containing buquinolate are stable overnight if stored in the refrigerator.
SUMMARY
A spectrophotofluorometric method developed for the determination of buquinolate in poultry tissue and eggs is sensitive to 0.1 p.p.m. Tissues were assayed from chickens following administration of the compound to broilers for 9-10 weeks in feed at the 75 and 100 gram per ton level, and to layers at 100 grams per ton of feed for 18 months. Levels of less than 0.1 to 0.39 p.p.m. were detected in liver, kidney, fat and skin at zero withdrawal, while muscle contained less than 0.1 p.p.m. at zero withdrawal. Tissues collected following 1 or 2 days withdrawal from buquinolate contained less than 0.1 p.p.m. with the exception of an occasional sample that contained levels of the order of 0.1 p.p.m. Eggs collected from layers following the administration of buquinolate at 100 grams per ton of feed for 18 months contained from 0.11 to 0.23 p.p.m. of buquinolate. REFERENCES Engle, A. T., R. P. Humphrey and C. Johnson, 1966. Buquinolate, a new broad spectrum coccidiostat. Poultry Sci. 4S: 1082. Herrett, R. J., G. M. Klein, C. W. Williams, and J. P. Heotis, 1966. The distribution of C" in the chick following the administration of Buquinolate-3-C". Poultry Sci. 45: 1092.
Calcium and Phosphorus in the Diet of Coturnix Quail1 B.
F.
MILLER
Department of Poultry Science, Colorado Agricultural Experiment Station, Fort Collins, Colo. 80521 (Received for publication October 7, 1966)
INTRODUCTION
T
HE Coturnix quail (Coturnlx coturnix japonica) provides a relatively new and increasingly used tool for re1
Published with the approval of the Director of the Colorado Agricultural Experiment Station as S. S. paper 1144, journal series.
search. Its characteristics and requirements are not well explored. Johnson (1960) in a review of the usefulness of the Coturnix emphasized that the quail has many advantages over other laboratory animals including resistance to disease and relatively small maintenance cost. This animal is easily handled, reproduces in captivity readily
CA AND P IN QUAIL DIETS
687
and requires little space and feed. Howes and Cassidy (1964b) found similar results and Frey (1961) have outlined a procedure when the study of phosphorus was confor obtaining and caring for the Coturnix tinued to eight weeks of age. Feed intake quail; while Wilson et al. (1961) have in- and body weight at eight weeks of age and vestigated reproductive capacities and ob- tibial bone ash at four weeks of age were tained data concerning fertility, hatchabili- improved in chicks receiving supplementy, growth rate and egg production. tary inorganic phosphorus. The National Research Council (1966) Turkey poults (Temperton and Cassirecommended one percent calcium and 0.6 dy, 1964c) fed an all-mash diet containing percent phosphorus in the diet of starting no feedstuffs of animal origin and with no and growing chicks. For starting poults the phosphorus supplement developed severe recommendations are increased to 1.2 per- rickets characterized by bowing of the leg cent calcium and 0.8 percent phosphorus. bones and beading of the ribs. Affected One percent phosphorus is recommended poults recovered and clinical symptoms disfor starting and growing pheasants, but no appeared as a result of adding sufficient sorecommendation is given in regard to dium phosphate to provide 0.9 percent young quail for either calcium or phospho- phosphorus in the diet. rus. For quail breeders, 2.3 percent calcium Temperton et al. (1965a) showed that and 1.0 percent phosphorus are recom- with light weight hybrid pullets, diets conmended. taining 0.45 percent total phosphorus and Sunde and Bird (1956) working with 0.14 percent available phosphorus during pheasant chicks reported that feeding a the first eight weeks did not result in any diet containing 0.66 percent phosphorus significant depression of body weight, feed (analytical analysis) resulted in high mor- consumption, feed efficiency or livability tality. The pheasant chicks after two weeks when compared to birds receiving diets on this diet showed a distinct bending of supplemented to the N.R.C. recommended the tarsometatarsus near the hock joint levels with inorganic phosphorus. Temperand at the proximal end of the tibiotarsus. ton et al. (1965b) further observed no Bone ash values were reduced from a nor- marked effects on total bone ash, calcium mal of 52 to 38 percent when the basal diet or phosphorus content of toes from feeding was fed. Additions of 0.3 to 0.8 percent of a diet containing 0.46 percent total phosphosphorus from dicalcium phosphate re- phorus and 0.14 percent available phosphosulted in normal growth, normal bone for- rus from one-day old to eight weeks of age. The amount of phosphorus in the toes at mation and calcification. Temperton and Cassidy (1964a) have 18 weeks of age was approximately oneshown that pullet chicks can grow normally half the amount of calcium in all treatup to 26 days of age receiving a vegetable ments. protein diet containing 0.48 percent total Plant source phosphorus (Vandepopuphosphorus and 0.23 percent non-phytin liere et al., 1961) was readily available to phosphorus. Balance studies indicated support growth when fed to broiler chicks these chicks retained approximately 60 per- at a calcium to phosphorus ratio of 1:1; cent of the phytin phosphorus ingested. however, it did not allow for increased The percent bone ash, calcium and phos- bone ash as did dicalcium phosphorus. phorus concentrations and calcium to phos- Chicks which had bone ash values of 28 phorus ratio of tibiae reflected normal bone percent did not exhibit symptoms of rickets formation. These same workers, Temperton when fed a diet containing 0.28 percent
688
B. F. MILLER
plant phosphorus and a calcium to phosphorus ratio of 1:1. A phosphorus level of 0.72 percent from dicalcium phosphate with a Ca:P ratio of 1:1 to 1.5:1 produced maximum growth in battery trials. The criterion, growth, appeared to be sufficient to determine the phosphorus adequacy of a diet when the level approached the requirement. It was essential to consider both calcium and phosphorus levels and the calcium to phosphorus ratio when determining phosphorus availability, irrespective of the phosphorus source. Nelson et al. (1964) reported that Coturnix quail breeders produced better when they received 2.5 and 3.0 percent calcium as compared to birds that received 1.0, 1.5 and 2.0 percent calcium in their diet. Little difference in egg production was found between groups receiving 0.6 and 0.8 percent phosphorus. There was little difference in fertility between treatments, but hatchability was variable with higher hatchability on the 2.5 and 3.0 percent calcium diets and on 0.8 percent phosphorus. Very little information appears in the literature about the nutritive requirements of the Coturnix quail. In view of the increasing acceptance of Coturnix as a laboratory animal and the apparent lack of information of their nutritional requirements, this study was initiated. In addition, it was hoped to add to the limited knowledge of the phosphorus and calcium requirements of Coturnix quail as well as add to the overall understanding of the role of phosphorus in the diet. PROCEDURE
Twelve pens of one-day old Coturnix quail with 15 chicks per pen were utilized in the first trial. The chicks were randomly distributed among the pens. The different treatments were randomly arranged within a modified Jamesway battery brooder. Modification consisted of covering
the floor and sides of each pen with onequarter inch mesh screen to prevent the chicks from escaping. The temperature of the brooders was adjusted to 37°C. during the first week of brooding. Room temperature was held at approximately 23°C. The brooder temperature was decreased 3°C. each week thereafter until room temperature was reached. The birds received 14 hours of light from fluorescent fixtures placed overhead in the room. The room was air conditioned and force ventilated. During the first two weeks, quail feeders were utilized to feed the chicks. One quart glass chick waterers were utilized for watering during this time. The chick waterers were modified by filling the lower half of the watering area with plastic cement to prevent the quail chicks from getting into the deep water and drowning. Feed was placed on paper towels during the first three days to encourage eating. After the first two weeks the standard battery feeders and waterers were utilized for the growing quail. Feed consumption was measured weekly. Wastage of feed during the first two weeks was excessive in some lots making these data unreliable. Feed wastage during the remainder of the experiment was slight and fairly uniform from one experimental group to another. The average body weights for each group were determined weekly by weighing the group and dividing by the number of birds within the group. Mortality was recorded daily; however, any birds that died during the first week of the experiment were replaced with comparable aged birds receiving the basal (control) diet. The basal diet (Table 1) had an analyzed calcium level of 0.44 percent and a phosphorus level of 0.59 percent. Ground limestone and dicalcium phosphate were the only ingredients added to increase the level of calcium and phosphorus for the various experimental diets. The different levels of calcium and phosphorus studied
689
CA AND P IN QUAIL DIETS
are shown in Table 3. A random sample of 10 birds from each group was individually scored for proper feathering with 10 indicating a perfect covering of feathers. At six weeks of age, when the experiment was terminated, one male and one female were sacrificed from each group, and the tibiae were analyzed for percent ash of moisture and fat-free bone. In the second trial, the same procedure was followed; however, each experimental group was replicated with 10 birds per experimental group. During the sixth week of the experiment all surviving birds were individually feather scored as before. Comparisons of average body weight, feed efficiency, percent bone ash, livability, and feather scores were made among the different treatments. Samples of the feed were analyzed for phosphorus using the molybovandophosphoric acid procedure with wet digestion in perchloric acid (Barton, 1948). Calcium was determined on the atomic absorption TABLE 1.—Composition of basal diet utilized to study phosphorus requirement of Coturnix quail Ingredient
Percent
Corn, ground yellow Soybean meal (50% protein) Alfalfa meal (20% protein) Added per 45.4 kg. of diet: Vitamin A Vitamin D 3 Vitamin E Vitamin K Thiamine Riboflavin Pyridoxine Pantothenic acid Folacin Niacin Biotin Vitamin B12 Choline Salt and trace mineral mix: NaCl, iodized Mg Mn Zn Fe Cu Mo Methionine
41. 5 51. 0 5. 0 400,000 100,000 1,000 50 100 200 100 400 40 3,000 4 0.5 20 136 200 50 50 25 3 2 50
I.U. I.U. I.U. mg. mg. mg. mg. mg. mg. mg. mg. mg. gm. gm. p.p.m. p.p.m. p.p.m. p.p.m. p.p.m. p.p.m. gm.
TABLE 2.—Comparison of mean weekly body weights and mean weekly body weights as compared to the weights reported by Wilson et al. {1961) Age of birds (weeks) 1 Mean body weight (gm.) (Trials 1 and 2) Mean body weights 1 (gm.) 1
,2
3
22 40 65 38 64 84
4
5
84 100
94 111
Wilson et al. (1961).
spectrophotometer after digestion.
the above wet
RESULTS AND DISCUSSION
The basal diet containing 0.44 percent calcium and 0.59 percent phosphorus on an analytical basis resulted in satisfactory growth and development (Table 3). Body weights and growth rate of the birds that received the basal diet were above the average for all treatment groups. Percent bone ash was not depressed to any significant extent by the low levels of calcium and phosphorus in the diet. Feather scores of these birds were the highest in all cases. The birds that received the basal diet showed no ill effects from consuming this diet. Livability was not affected by the diet. Feathering was significantly enhanced by this treatment over some of the other diets utilized in the study. Statistical analysis of mean body weights indicated there were no significant differences among treatments. Average body weights and Ca:P ratios of the diets used are presented in Table 3. The greatest body weight was found where the birds received 0.70 percent Ca and 0.58 percent P in the diet. Mean body weights (Table 2) were compared by weeks to those reported by Wilson et al. (1961). The birds in this experiment were approximately one week behind in their weights. These birds were slow in getting started as indicated by their one-week old weights. They reached sexual maturity at about five to six weeks of age,
690
B. F. MILLER
TABLE 3.—Effects of dietary calcium and phosphorus levels upon quail body weights and percent bone ash at six weeks of age1
Diet
Calcium2 percent
11 (Basal) 21 14 18 19 12 20 15 13 22 16 17
0.44 1.12 0.90 1.08 1.29 0.70 1.14 1.41 1.08 2.30 1.58 2.18
Phosphorus percent2
Ca:P ratio
Average body wt.* (grams)
Percent bone ash
0.59 1.18 0.83 1.02 1.14 0.58 0.86 0.76 0.58 1.14 0.74 0.75
0.7:1 1.0:1 1.1:1 1.1:1 1.1:1 1.2:1 1.3:1 1.8:1 1.9:1 2.0:1 2.1:1 2.9:1
101 98 101 99 96 103 98 98 97 101 98 100
28 31 31 32 31 28 31 29 30 30 30 31
1 Combined body weights of birds in Trials 1 and 2. ! Calcium and phosphorus level determined by chemical analysis of diet 3 No significant difference between body weights at the 5 percent level.
similar to results reported by Wilson et al. (1961). The birds that were fed diet 12 were heaviest, averaging 103.2 gm. (Table 3) on a diet containing 0.7 percent calcium and 0.58 percent phosphorus. Their feathering was comparable to the controls in trial 2 (Table 5) and somewhat depressed in trial 1 (Table 4). Tables 4 and 5 present the analysis of variance of the mean feather scores during trials 1 and 2, respectively. When the multiple range test (Duncan, 1955) was applied to the treatment means in both exper-
iments, the basal diet (0.7:1 Ca:P ratio) and diet 14 (1.1:1 Ca:P ratio) consistently ranked in the highest group although they are not significantly better than all treatments. Birds receiving diets 13, 15, 16, 17, 20 and 22 (1.3:1 to 2.9:1 Ca:P ratios) were significantly not as well feathered as those birds receiving the basal diet or diet 14 (0.90% Ca and 0.83% P ) . Birds receiving diet 22 (2.3 Ca and 1.1 P) were consistently low in feather score, while the other groups showed considerable variation. Analysis of the feather scores verified the visual observations made during the experiment. Figure 1 provides a comparison of the typical feathering of a bird that received the basal diet and one that received diet 20. Analysis of the data on bone ash indicated there was no significance between treatments (Table 3). The females had a slightly higher bone ash content than the males. The average bone ash content of fat-free tibia was 29.8 percent with a range of 25.8 to 37.0 percent. No significant differences were found in feed efficiency between the various levels of calcium and phosphorus. None of the birds died after the first week of age, except accidental deaths during handling. All diets
TABLE 4.—Analysis of variance of mean feather scores in Trial 1 Sources
D.F.
S.S.
M.S.
F.
129.9 19.0 30.S
4.26 .62
Treatment Period Error
11 1 11
1,429 19 346
Total
23
1,894
Duncan's test of mean feather scores, Trial 1 } Treatment 11 14 19 16 12 (Basal) Avg. 9.4 9.3 9.2 8.2 8.1
Sig.
* N.S.
17
21
20
22
18
IS
13
8.0
8.0
7.9
7.8
7.S
7.3
6.9
* Significant at the five percent level. N.S. Not significant. Any groups underlined by the same line are not statistically different from each other at the P<0.05 level. 1
CA AND P IN QUAIL DIETS
691
TABLE 5.—Analysis of variance of individual bird feather scores at six weeks of age in Trial 2 Sources
D.F.
s.s.
M.S.
Treatment Error
11 203
151
13.73 .96
Total
214
345
19-4
Duncan's test of mean feather scores, Trial 2.1 Treatment 11 14 12 18 21 (Basal) Avg. 9.8 9.4 9.3 9.0 8.9
Sig. 14.30
IS
10
19
13
22
20
17
8.3
8.3
8.3
8.2
7.9
7.1
6.8
** Significant at the one percent level. Any groups underlined by the same line are not statistically different from each other at the P<0.01 level. 1
were capable of sustaining the growing quail to maturity (six weeks of age). SUMMARY One-day old Coturnix quail chicks were reared to maturity (six weeks of age) on
4 f \
Bfc M& '^..
FIG. 1. The bird on the left received the basal diet containing 0.44 percent calcium and 0.59 percent phosphorus while the bird on the right received a similar diet containing 1.14 percent calcium and 0.86 percent phosphorus (diet 20).
an all-vegetable diet containing various calculated levels of calcium and phosphorus. The basal diet contained 0.44 percent calcium and 0.59 percent phosphorus on an analytical basis. Calcium and phosphorus content of the different diets was modified by adding ground limestone and dicalcium phosphate to the basal for the different levels and ratios of calcium and phosphorus studied. The basal diet containing 0.44 percent calcium and 0.59 percent phosphorus resulted in satisfactory gains in body weight. There was no evident impairment of bone ash in birds receiving this diet. Birds that received diets with 0.7:1 and 1.1:1 calcium to phosphorus ratios in general showed significantly better feathering than birds fed diets with 2:1 or higher Ca:P ratios. This was especially true when levels of calcium and phosphorus reached two percent and one percent, respectively. There was no significant difference in feed efficiency or livability between the various diets. Coturnix quail chicks are capable of utilizing an all-vegetable diet that is low in calcium and phosphorus providing it is adequately supplied with vitamins and other minerals. The level of calcium and phosphorus in
692
B. F. MILLER
the diet and the Ca:P ratio had a significant effect on the feathering of growing quail. As the calcium and phosphorus content of the diet increased, the relative feathering of the birds decreased. A spreading of the Ca:P ratio also caused a similar depression in feathering. ACKNOWLEDGMENTS
This investigation was supported in part by a grant-in-aid from the Faculty Improvement Committee, Colorado State University, to whom the author is grateful. Appreciation is expressed to The Ray Ewing Company Division of HoffmannLaRoche, Inc., Pasadena, California, for supplying the vitamin supplements for this study. The assistance of Dr. Eldon W. Kienholz and Dr. Cheryl F. Nockels during the conduct of this study and preparation of the manuscript has been very helpful. REFERENCES Barton, C. J., 1948. Photometric analysis of phosphoric rock. Anal. Chem. 20: 1068-1073. Duncan, D. B., 19SS. Multiple range and multiple F tests. Biometrics, 11: 1^2. Howes, J. R., and W. D. Frey, 1961. Coturnix quail for avian research. Feedstuffs, 33, (21) May 27, 1961. Johnson, E. L., 1960. A new research tool . . . Coturnix quail. Minnesota Farm and Home Science, 18(1): 1. National Research Council of America, 1966. Nutrient requirements of poultry. National Acad-
emy of Sciences-National Research Council, Washington, D.C. Publication 1345. Nelson, F. E., J. K. Lauber and L. Mirosh, 1964. Calcium and phosphorus requirement for the breeding Coturnix quail. Poultry Sci. 43: 1346. Sunde, M. L., and H. R. Bird, 1956. A critical need of phosphorus for the young pheasant. Poultry Sci. 35: 424-^30. Temperton, H., and J. Cassidy, 1964a. Phosphorus requirements of poultry. I. The utilization of phytin phosphorus by the chicks as indicated by balance experiments. British Poultry Sci. 5: 75-80. Temperton, H., and J. Cassidy, 1964b. II. The utilization of phytin phosphorus by the chick for growth and bone formation. British Poultry Sci. S: 81-86. Temperton, H., and J. Cassidy, 1964c. III. The effect of feeding a vegetable-type diet without supplementary phosphorus to turkey poults. British Poultry Sci. 5: 87-88. Temperton, H., F. J. Dudley and G. J. Pickering, 1965a. Phosphorus requirements of poultry. IV. The effects on growing pullets of feeding diets containing no animal protein or supplementary phosphorus. British Poultry Sci. 6: 125-133. Temperton, H., F. J. Dudley and G. J. Pickering, 1965b. The phosphorus requirements of growing pullets between 8 and 18 weeks of age. British Poultry Sci. 6: 143-152. Vandepopuliere, J. M., C. B. Ammermon and R. H. Harms, 1961. The relationship of calciumphosphorus ratios on the utilization of plant and inorganic phosphorus by the chick. Poultry Sci. 40: 951-957. Wilson, W. O., U. K. Abbott and H. Abplanalp, 1961. Evaluation of Coturnix (Japanese quail) as pilot animal for poultry. Poultry Sci. 40: 651-657.
NEWS AND NOTES (continued from page 650) sociation are: President—E. E. Penionzhkevich (U.S.S.R.); Hon. Past Presidents—H. H. Alp (U.S.A.), R. C. Blake (Australia), R. Coles (England), A. Ghigi (Italy), W. D. Termohlen (U.S.A.), A. Wiltzer (France); Vice-Presidents—Z. BenAdam (Israel), W. P. Blount (England), R. Pero (France); Hon. Past Vice-Presidents—H. Engler (Switzerland), C. S. Th. Van Gink (Netherlands); W. P. S. A. NOTES Treasurer and Assistant Secretary—R. G. Jaap The officers of the World's Poultry Science As(U.S.A.); Secretary and Assistant Treasurer—I. (continued < n page 754)
stories of how several companies who are now leaders in the production of further processed poultry and egg products got that way. Further information can be obtained by writing to Dr. George Mountney, Department of Poultry Science, Ohio State University, 674 West Lane Avenue, Columbus, Ohio 43210.