- Email: [email protected]
Hematological and Growth Response of Japanese Quail Fed an Iron-Copper Deficient Diet
RAW SOYBEAN DIETS N.R.C. 6th revised edition. National Academy of Sciences, Washington, D.C. Nesheim, M. C , J. D. Garlich and D. T. Hopkins, 1962. Studies on the effect of raw soybean meal on fat absorption in young chicks. J. Nutrition, 78: 89. Nesheim, M. C , 1967. Feeding replacement pullets. Feedstuffs, 39(18): 39. Rogler, J. C , and C. W. Carrick, 1964. Studies on raw and heated unextracted soybeans for layers. Poultry Sci. 43: 605-612. Salman, A. J., and J. McGinnis, 1968. Effect of supplementing raw soybean meal with methionine on performance of layers. Poultry Sci. 47: 247-251. Saxena, H. C , L. S. Jensen and J. McGinnis, 1963a.
2101
Influence of age on utilization of raw soybean meal by chickens. J. Nutrition, 80: 391-396. Saxena, H. C , L. S. Jensen, J. V. Spencer and J. McGinnis, 1963b. Production, interior egg quality and some physiological effects of feeding raw soybean meal to laying hens. Poultry Sci. 42: 291-293. Snedecor, G. W., and W. G. Cochran, 1967. Statistical Methods. 6th edition. Iowa State Univ. Press, Ames, Iowa. Summers, J. D., J. D. McConachie, S. J. Slinger and W. F. Pepper, 1966. The value of raw unextracted soybeans for laying hens. Poultry Sci. 45: 165-169. Waldroup, P. W., D. R. Sloan andR. F. Davenport, 1969. The use of raw and extruded soybeans in layer diets. Poultry Sci. 48: 1481-1486.
K. W. WASHBURN AND R. W. LOWE1
Department of Poultry Science, University of Georgia, Athens, Georgia 30602 (Received for publication February 25, 1974)
ABSTRACT The effects of iron, copper deficient skim-milk diets on hematology and growth of Japanese quail were determined. The feeding of iron, copper deficient diets resulted in rapid and severe reductions in packed cell volume, hemoglobin concentration, mean cell hemoglobin concentration and rate of growth, although livability was only slightly affected. Addition of iron and copper to the skim-milk diet only partially alleviated its adverse effects on hematology and growth. POULTRY SCIENCE 53: 2101-2107, 1974
T
HE Japanese quail (Cotumix coturnix japofiica) is used extensively as a pilot animal in nutrition, genetic and physiology studies. In genetic studies, one use has been in selection experiments for hematological response to iron, copper deficient diets (Stino, 1971). However, information is meager on what constitutes adequate dietary Fe, Cu in quail rations and the extent of the hematological response to Fe, Cu deficiencies. Harland era/. (1973), using an EDTA extracted soya-protein diet, reported that the Fe and Cu requirements of young Japanese
1. Present address: County Office Building, Homer, Georgia 30547
quail were 90-120 and 5 p.p.m., respectively. Atwal et al. (1964), in a study of the hematology from hatching to maturity, found that the erythrocyte counts, hematocrit, and hemoglobin concentration of both sexes increased with age. After 43 days of age, a significant increase in erythrocyte counts, hematocrit, and hemoglobin concentration was noted in the males. The hematocrit values ranged from 22-43% for the 1-43 day age period. Erythrocyte counts ranged from 2.55.2 million /mm. with a range in hemoglobin concentration of 7.6-15.3 gms./lOO cc. blood. The present study is concerned with the effects of a Fe, Cu deficient skim-milk diet on the hematology and growth of immature Japanese quail.
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Hematological and Growth Response of Japanese Quail Fed an Iron-Copper Deficient Diet
2102
K. W. WASHBURN AND R. H. LOWE
The dietary treatments used in Trial 1 consisted of a corn-soya broiler starter (Table 1) which contained 250-200 p.p.m. Fe and 10-15 p.p.m. Cu and a skim-milk diet which contained 7-10 p.p.m. Fe and 2.5-5 p.p.m. Cu. The skim-milk diet was similar to that described by Hill and Matrone (1961) and later used by Washburn (1969) to study genetic differences of chickens in response to Fe, Cu deficiencies. In Trial 2 four dietary treatments were used: 1) a commercially available corn-soya
MATERIALS AND METHODS The hematological and growth response of Japanese quail to an iron, copper deficient diet was studied in two trials. In both trials a population of Japanese quail maintained at the Southern Regional Poultry Genetics Laboratory (Marks, 1967) was used. All quail were maintained in Beacon #B735 quail starting batteries and at 14 days of age were wingbanded and randomized to dietary treatment groups which were fed ad libitum.
Corn soya Ingredients Ground yellow corn Soybean meal (dehulled) Fish meal Poultry by-product meal Fat Dehydrated alfalfa (17%) Ground limestone Defluorinated phosphate Salt Methionine (DL) Trace mineral mix Vitamin premix
Skim milk
Broiler
Turkey
%
%
55.30 32.25
42.00 40.90 5.00 5.00 2.50 1.50 1.00 1.50 0.40 0.10 0.103
— 5.00 2.50 1.50 0.75 1.85 0.40 0.10 0.10' 0.252 100.00
4
Ingredients Corn starch Dried skim milk Methionine (dl) Arginine Glycine Choline Chloride (35%) Corn oil NaCl KK.PO,, MgC0 3 Na2HP04 CaHP0 4 CaC0 3 Mineral mix5 Vitamin mix 6
% 31.50 59.00 0.49 0.69 0.69 0.59 3.92 0.33 1.08 0.25 1.00 0.28 0.18
100.00
'Trace mineral mix contained (%): Ca, 26.0; Mn, 6.0; Co, 0.04; Fe, 2.5; Cu, 0.2; Zn, 2.0; I, 0.12. 2 Vitamin premix provided (per kg./diet): vitamin A, 4400 I.U.; vit. D3, 880 I.C.U.; vit. E, 11 I.U.; riboflavin, 4.4 mg.; Ca pantothenate, 9.6 mg.; nicotinic acid, 44 mg.; choline CI, 220 mg.; vit. B12, 6.6 meg.; vit. B6, 2.2 mg.; menadione sodium bisulfite, 3.49 mg.; folic acid, 55 mg.; d-biotin, 11 mg.; thiamine mononitrate, 2.2 mg.; ethoxyquin, 125 mg. 3 Trace mineral mix provides (in feed): 600 p.p.m. Mn, 0.4 p.p.m. Co, 25 p.p.m. Fe, 2 p.p.m. Cu, 20 p.p.m. Zn, 1.2 p.p.m. I. 4 Vit. premix provides (per kg. of feed): 11023 IJJ. vitamin A, 1764 I.C.U. vitamin D3, 11 I.U. vitamin E, 6.6 meg. vitamin B12, 4.4 mg. riboflavin, 8.8 mg. calcium pantothenate, 44.1 mg. niacin, 220.5 mg. choline CI, 0.9 mg. folic acid, 2.2 mg. thiamine HC1, .44 mg. biotin, 2.2 mg. menadione S. B., 125.7 mg. ethoxyquin, 4.4 mg. procain penicillin. 5 The mineral mixture supplied the following expressed as milligram per kg. diet: Zn, 100; Mn, 70; I, 0.5; Mo, 5.0; Se, 0.2. 6 The vitamin mixture supplied the following (in units or milligrams per kg. diet): vitamin A, 5300 I.U.; vitamin D3, 400 I.C.U.; thiamine HC1, 3.5; riboflavin, 5.7; Ca pantothenate, 18.5; pyridoxine, HC1, 2.2; niacin, 53; folic acid, 1.5; biotin, .7; B l2 , .018; vitamin K, .8; vitamin E, 22.
Downloaded from http://ps.oxfordjournals.org/ by guest on April 10, 2015
TABLE 1.—Composition of experimental diets used
soya soya + Fe milk milk + Fe, Cu
— —
10.5±2 10.6±.2 9.2±.2 10.1±.2
3 wks.
366±7 330±5
299 ±6 319±7 309±7 302±7
3 wks.
396±6 303 ±9
366±7 368±9 262±12 330±8
9.8 5.5
Trial 1
11.5±.2 10.1 ±.2 6.6±.3 9.2+.2
Trial 2
4 wks.
Age
11.3 4.2
11.8+.4 11.2±.4 5.1±.3 8.9±.4
5 wks.
.05).
Hemoglobin (gms./lOO cc. blood)
352+8 297±9
Trial 1
336±5 332±7 290±9 319+7
Trial 2
Those means with different superscripts were significantly different (P : —data not obtained.
Corn soya Skim milk
Corn Corn Skim Skim
Diet
soya soya + Fe milk milk + Fe, Cu
Corn soya Skim milk
Corn Corn Skim Skim
Diet
RBC counts (x: 10,000) Age 5 wks. 4 wks.
10.5a 4.8 b
10.9s 10.6" 6.9" 9.4C
X
371° 310b
333a 340 s 287" 317c
X
— —
29.4±.4 28.6±.4 27.0±.5 28.5±.5
3 wks.
131 + 1 120+2
120+.9 117+.9 110+1 117+1
3 wks.
TABLE 2.—Hematological response of Cotumix quail fed iron, copper defici
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2104
K. W. WASHBURN AND R. H. LOWE 48
/
44
/
40
^ ^ < ee
\
36 32
t-
28 24 20
'
> u
2
3 4 AGE (WEEKS)
'
'
2
3 4 AGE (WEEKS)
5
48 44 40 o
36
<
32
"
28 24 20
FIG. 1. Effect of diets on PCV in Coturnix quail. % % Corn-Soya %—0 Corn-Soya + lOOp.p.m. Fe O O Skim Milk O—O Skim Milk + 100 p.p.m. Fe, 5 p.p.m. Cu
RESULTS AND DISCUSSION The packed erythrocyte volume (PCV) response to the skim-milk diet deficient in Fe, Cu was rapid and severe in both trials (Fig. 1) with statistically significant differences being observed one week after initiation. After one week, the PCV of groups in Trial 1 receiving the Fe, Cu deficient diet was 37% compared to 47% for controls receiving the diets adequate in Fe, Cu. A less rapid reduction was observed in Trial 2. After one week
the PCV of the deficient groups in Trial 2 was reduced to 34% compared to 36% for controls receiving the corn-soya turkey starter. The rapid decline in PCV after initiating the deficient diet indicates that Japanese quail are sensitive to deficiencies of this type. The extremely fast growth rate of quail at this age would rapidly dilute the Fe, Cu reserves
Downloaded from http://ps.oxfordjournals.org/ by guest on April 10, 2015
turkey starter (Table 1) containing 250-300 p.p.m. Fe and 10-15 p.p.m. Cu and similar in composition to the corn-soya diet used in Trial 1, but which appeared to give superior growth; 2) the turkey starter to which 100 p.p.m. Fe (as FeS0 4 ) was added; 3) the skim-milk diet used in Trial 1; 4) the skim milk diet to which 100 p.p.m. Fe (as FeS0 4 ) and 5 p.p.m. Cu (as CuS0 4 ) was added. The basal corn-soya diets in each trial were fed until 2 weeks of age, at which time the quail were placed on experimental dietary treatments for a 3-week period. Body weights and packed erythrocyte volume (by the micro-hematocrit method) were obtained at weekly intervals from 2-5 weeks of age for both trials. Feed consumption was measured in Trial 2. Erythrocyte counts, mean cell volumes, hemoglobin concentration and mean cell hemoglobin concentration were determined at intervals shown in Table 2. Erythrocyte counts and mean cell volume were obtained by a model F Coulter Counter and hemoglobin concentration by the acidhematin method of Dennington and Lucas (1955). Two replicates were used for each dietary treatment in each trial with 20 quail used per replicate in Trial 1 and 12 per replicate in Trial 2. Data of two quail from some subsets of Trial 1 and of one quail in Trial 2 were deleted to equalize subclass numbers for statistical analysis by analysis of variance and Duncan's separation of means (Li, 1957).
2105
IRON-COPPER DEFICIENCY IN QUAIL
Further rapid decreases were noted in PCV of quail receiving the skim-milk diet in both trials so that after three weeks the PCV of the Fe, Cu deficient groups in Trial 1 had decreased to 20% compared to 37% in the control group receiving the corn-soya diet. In Trial 2 the PCV of the deficient group decreased to 22% compared to 42% in the control groups receiving the corn-soya diet. Even with this severe reduction in PCV, mortality was slight. In Trial 1, 3/40 quail on Fe, Cu deficient diets and in Trial 2, 1/24 had died by the end of the 5-week experimental period. The addition of 100 p.p.m. Fe to the turkey corn-soya diet in Trial 2 (Fig. 1) did not result in a change in PCV indicating that the amount of Fe in the diet (250-300 p.p.m. Fe) was adequate for maximum hematopoeisis. The addition of Fe and Cu to the skim-milk resulted in a significant elevation of PCV above that of groups receiving the basal skim-milk diet. However, the PCV did not approach the level obtained when the turkey
corn-soya diets were fed. Although the establishment of Fe requirements was not designed into this experiment, this would suggest that the 90-100 p.p.m. Fe requirement for quail suggested by Harland et al. (1973) may not be adequate for normal hematology. Harland et al. (1973) used EDTA extracted soya to produce an Fe deficient diet while in the
110 100
90 80
I 70 < 50 £ t. &
40h 301-
110 ~ 100 S*
90 80
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tt 60 t-
//
V
// js
J
-°
*' ''\^^^
ft/
// Of
50 40 30 i
'
i
3 4 AGE (WEEKS)
FIG. 2. Effect of diets on growth in Coturnix quail. # % Corn-Soya •----# Corn-Soya +- 100 p.p.m. Fe O O Skim Milk O—O Skim Milk + 100 p.p.m. Fe, 5 p.p.m. Cu
Downloaded from http://ps.oxfordjournals.org/ by guest on April 10, 2015
so that any deficiencies of nutrients necessary for hematopoeisis would quickly be evident. However, the hematology of the quail apparently is not as responsive to chemicals that may have a detrimental effect. Ernst and Ringer (1968) in an evaluation of the effects of pesticide chemicals on erythrocyte of Japanese quail reported that continuous dietary administration of DDT, Zectron or Zytron significantly reduced the PCV and total erythrocyte counts of blood from mature quail but not in immature quail. They concluded that PCV was not a useful criterion of pesticide toxicity since near lethal levels were necessary to cause a significant change in PCV. In contrast to the rapid response observed in quail in this study, Washburn (1969) found that chickens placed on an Fe, Cu deficient diet following feeding of a normal diet for their first two weeks required at least three weeks on the deficient diet to show even a small reduction in PCV.
2106
K. W. WASHBURN AND R. H. LOWE
These results show evidence of the typical hypochromic, microcytic anemia expected from a severe Fe, Cu deficiency. The relative differences in RBC counts, MCV, hemoglo-
TABLE 3.- -Feed conversions (gms. feed/gm.
Age (days) days on diet
bin and MCHC of the groups in Trial 1 and Trial 2 receiving the skim milk diet and the corn-soya diets paralleled differences observed in PCV. The addition of 100 p.p.m. Fe to the corn-soya diet in Trial 2 did not significantly alter the hematological picture. The groups in Trial 2 which were fed the skim-milk diet to which 100 p.p.m. Fe and 5 p.p.m. Cu was added also exhibited a hypochromic, microcytic anemia but the depression in RBC counts, MCV, hemoglobin content and MCHC were much less severe than the groups receiving the skim-milk diet without additional Fe, Cu. The effect of the various diets on growth is presented in Figure 2. The feeding of the Fe, Cu deficient skim-milk diet resulted in approximately 30% reduction in body weight by 5 weeks. This depression in growth may not be primarily attributed to the Fe, Cu deficiency since when quail in Trial 2 were given the skim milk diet with 100 p.p.m. Fe, 5 p.p.m. Cu growth did not approach that of the groups fed the corn-soya diet. Feed conversion ratio (Table 3) was significantly
gain) of Coturnix fed iron-copper deficient or adequate diets (Trial 2) 28-35 14-21
14-35 0-21
3.1 3.2 3.1
21- •28 7- 14 4.0 3.9 4.0
6.6 7.2 6.9
(292) (273) 4.7 a
X
2.7 3.1 2.9
4.1 4.1 4.1
6.5 6.6 6.6
(262) (271) 4.5 a
Skim milk
R, R7 X
3.8 3.8 3.8
5.3 5.0 5.2
9.6 11.1 10.4
(198) (199) 6.5 b
Skim milk + Fe, Cu
R, R, X
3.7 3.9 3.8
5.3 4.4 4.8
5.6 5.4 5.5
(223) (230) 4.7 a
Corn-soya
14-21 0-7 R, R?
X Corn-soya + Fe
R, R?
a
R, = replicate 1; Rj = replicate 2. ( ) = feed consumption. Those means with different superscripts were significantly different (P :
.05).
Downloaded from http://ps.oxfordjournals.org/ by guest on April 10, 2015
present study a skim-milk diet was used. The effects of the various diets on blood measurements other than PCV are presented in Table 2. In Trial 1 after 5 weeks the RBC counts of the groups receiving the skim milk diet were 3.03 million compared to 3.96 million for the groups receiving the corn-soya diet. The MCV was 66 jx-3 compared to 96 (x.3, the hemoglobin content was 4.2 gms. compared to 11.3 gms. and the MCHC was 20.4% compared to 29.1%. Similar changes were observed in Trial 2, the RBC counts of the groups receiving the skim-milk diet were 2.62 million compared to 3.66 million; the MCV was 85JA,.3 compared to 116u.. 3 ; the hemoglobin content was 5.1 gms. compared to 11.8 gms., and the MCHC was 22.9% compared to 27.6%. All these differences were statistically significant as were the differences in mean values for the experimental period.
IRON-COPPER DEFICIENCY IN Q U A I L
1964. Hematology of Coturnix from birth to maturity. Poultry Sci. 43: 1392-1401. Blair, R., M. L. Scott and R. J. Young, 1972. Unidentified factor activities in whole soybeans required for optimum growth of Coturnix. J. Nutr. 102: 1529-1541. Dennington, E. M., and A. M. Lucas, 1955. Blood techniques for chickens. Poultry Sci. 34: 360-368. Ernst, R. A., and R. K. Ringer, 1968. The effect of DDT, Zectran and Zytron on the packed cell volume, total erythrocyte count and mean corpuscular volume of Japanese quail. Poultry Sci. 47: 639-643. Harland, B. F., B. E. Fry, R. M. Jacobs and M. R. S. Fox, 1973. Mineral requirements of young Japanese quail. Federation Proc. 32: 930. Hill, C. H., and C. Matrone, 1961. Studies on copper and iron deficiencies of growing chickens. J. Nutr. 73: 425-431. Jacobs, R. M., M. R. Spivey Foxand M. H. Aldridge, 1969. Changes in plasma proteins associated with the anemia produced by dietary cadmium in Japanese quail. J. Nutr. 99: 119-128. Li,C. R. J., 1957. Introduction to Statistical Inference. Edwards Bros., Inc., Ann Arbor, Michigan. Marks, H. L., 1967. A Japanese quail control population. The Quail Quarterly, 4(1): 2-4. Stino, F. K. R., 1971. Divergent selection for packed erythrocyte volume in the Japanese quail under two environments. Ph.D. Thesis, Univ. of Georgia, Athens. Washburn, K. W., 1969. Hematological response of different stocks of chickens to iron, copper deficient diet. Poultry Sci. 48: 204-209. Wentworth, B. C , 1970. Sterility and reproductive inhibition of Japanese quail induced by mestranol ingestion. Poultry Sci. 49: 1477-1484.
REFERENCES Atwal, O. S., L. Z. McFarland and W. O. WUson,
NEWS AND NOTES (Continued from page 2085) received a Ph.D. from Louisiana State University, majoring in poultry nutrition. He was winner of the Charles Upp Award for outstanding graduate work in Poultry Science at L.S.U., and will continue to serve as Valmac's chief nutritionist.
Before joining Valmac, he was Poultry Nutritionist with Pillsbury Farms and Manager of the poultry research farm at L.S.U. Valmac Industries is one of the nation's large poultry processors with four plants in Arkansas at Russellville, Dardanelle, Pine Bluff and Waldrop.
(Continued on page 2136)
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increased in the groups fed the milk diet without supplemental iron, but conversion was not different in the groups receiving the skim-milk diet supplemented with Fe and Cu from groups receiving the corn-soya diet. The results of Blair et al. (1972) suggest that quail chicks fed an amino acid deficient diet require certain peptides for maximizing growth. When isolated soy protein was substituted for a portion of the amino acids in a basal amino acid diet, growth was increased by 35% and by 54% when heated ground soybeans were used. This apparent requirement for peptides obtainable from soy protein is a logical explanation for the growth depression of the quail given the semi-purified skim-milk diet. A number of factors have been shown to affect the hematology of quail; however, there is no evidence that rate of growth per se has any relationship. Wentworth (1970), in a study of the effects of mestranol ingestion on sterility and reproduction in Japanese quail, reported that both castration and mestranol ingestion significantly depressed PCV in sexually mature males. Jacobs et al. (1969) produced an anemia in Japanese quail by use of dietary cadmium. This anemia was associated with increased levels of plasma transferrins.
2107
2101
Influence of age on utilization of raw soybean meal by chickens. J. Nutrition, 80: 391-396. Saxena, H. C , L. S. Jensen, J. V. Spencer and J. McGinnis, 1963b. Production, interior egg quality and some physiological effects of feeding raw soybean meal to laying hens. Poultry Sci. 42: 291-293. Snedecor, G. W., and W. G. Cochran, 1967. Statistical Methods. 6th edition. Iowa State Univ. Press, Ames, Iowa. Summers, J. D., J. D. McConachie, S. J. Slinger and W. F. Pepper, 1966. The value of raw unextracted soybeans for laying hens. Poultry Sci. 45: 165-169. Waldroup, P. W., D. R. Sloan andR. F. Davenport, 1969. The use of raw and extruded soybeans in layer diets. Poultry Sci. 48: 1481-1486.
K. W. WASHBURN AND R. W. LOWE1
Department of Poultry Science, University of Georgia, Athens, Georgia 30602 (Received for publication February 25, 1974)
ABSTRACT The effects of iron, copper deficient skim-milk diets on hematology and growth of Japanese quail were determined. The feeding of iron, copper deficient diets resulted in rapid and severe reductions in packed cell volume, hemoglobin concentration, mean cell hemoglobin concentration and rate of growth, although livability was only slightly affected. Addition of iron and copper to the skim-milk diet only partially alleviated its adverse effects on hematology and growth. POULTRY SCIENCE 53: 2101-2107, 1974
T
HE Japanese quail (Cotumix coturnix japofiica) is used extensively as a pilot animal in nutrition, genetic and physiology studies. In genetic studies, one use has been in selection experiments for hematological response to iron, copper deficient diets (Stino, 1971). However, information is meager on what constitutes adequate dietary Fe, Cu in quail rations and the extent of the hematological response to Fe, Cu deficiencies. Harland era/. (1973), using an EDTA extracted soya-protein diet, reported that the Fe and Cu requirements of young Japanese
1. Present address: County Office Building, Homer, Georgia 30547
quail were 90-120 and 5 p.p.m., respectively. Atwal et al. (1964), in a study of the hematology from hatching to maturity, found that the erythrocyte counts, hematocrit, and hemoglobin concentration of both sexes increased with age. After 43 days of age, a significant increase in erythrocyte counts, hematocrit, and hemoglobin concentration was noted in the males. The hematocrit values ranged from 22-43% for the 1-43 day age period. Erythrocyte counts ranged from 2.55.2 million /mm. with a range in hemoglobin concentration of 7.6-15.3 gms./lOO cc. blood. The present study is concerned with the effects of a Fe, Cu deficient skim-milk diet on the hematology and growth of immature Japanese quail.
Downloaded from http://ps.oxfordjournals.org/ by guest on April 10, 2015
Hematological and Growth Response of Japanese Quail Fed an Iron-Copper Deficient Diet
2102
K. W. WASHBURN AND R. H. LOWE
The dietary treatments used in Trial 1 consisted of a corn-soya broiler starter (Table 1) which contained 250-200 p.p.m. Fe and 10-15 p.p.m. Cu and a skim-milk diet which contained 7-10 p.p.m. Fe and 2.5-5 p.p.m. Cu. The skim-milk diet was similar to that described by Hill and Matrone (1961) and later used by Washburn (1969) to study genetic differences of chickens in response to Fe, Cu deficiencies. In Trial 2 four dietary treatments were used: 1) a commercially available corn-soya
MATERIALS AND METHODS The hematological and growth response of Japanese quail to an iron, copper deficient diet was studied in two trials. In both trials a population of Japanese quail maintained at the Southern Regional Poultry Genetics Laboratory (Marks, 1967) was used. All quail were maintained in Beacon #B735 quail starting batteries and at 14 days of age were wingbanded and randomized to dietary treatment groups which were fed ad libitum.
Corn soya Ingredients Ground yellow corn Soybean meal (dehulled) Fish meal Poultry by-product meal Fat Dehydrated alfalfa (17%) Ground limestone Defluorinated phosphate Salt Methionine (DL) Trace mineral mix Vitamin premix
Skim milk
Broiler
Turkey
%
%
55.30 32.25
42.00 40.90 5.00 5.00 2.50 1.50 1.00 1.50 0.40 0.10 0.103
— 5.00 2.50 1.50 0.75 1.85 0.40 0.10 0.10' 0.252 100.00
4
Ingredients Corn starch Dried skim milk Methionine (dl) Arginine Glycine Choline Chloride (35%) Corn oil NaCl KK.PO,, MgC0 3 Na2HP04 CaHP0 4 CaC0 3 Mineral mix5 Vitamin mix 6
% 31.50 59.00 0.49 0.69 0.69 0.59 3.92 0.33 1.08 0.25 1.00 0.28 0.18
100.00
'Trace mineral mix contained (%): Ca, 26.0; Mn, 6.0; Co, 0.04; Fe, 2.5; Cu, 0.2; Zn, 2.0; I, 0.12. 2 Vitamin premix provided (per kg./diet): vitamin A, 4400 I.U.; vit. D3, 880 I.C.U.; vit. E, 11 I.U.; riboflavin, 4.4 mg.; Ca pantothenate, 9.6 mg.; nicotinic acid, 44 mg.; choline CI, 220 mg.; vit. B12, 6.6 meg.; vit. B6, 2.2 mg.; menadione sodium bisulfite, 3.49 mg.; folic acid, 55 mg.; d-biotin, 11 mg.; thiamine mononitrate, 2.2 mg.; ethoxyquin, 125 mg. 3 Trace mineral mix provides (in feed): 600 p.p.m. Mn, 0.4 p.p.m. Co, 25 p.p.m. Fe, 2 p.p.m. Cu, 20 p.p.m. Zn, 1.2 p.p.m. I. 4 Vit. premix provides (per kg. of feed): 11023 IJJ. vitamin A, 1764 I.C.U. vitamin D3, 11 I.U. vitamin E, 6.6 meg. vitamin B12, 4.4 mg. riboflavin, 8.8 mg. calcium pantothenate, 44.1 mg. niacin, 220.5 mg. choline CI, 0.9 mg. folic acid, 2.2 mg. thiamine HC1, .44 mg. biotin, 2.2 mg. menadione S. B., 125.7 mg. ethoxyquin, 4.4 mg. procain penicillin. 5 The mineral mixture supplied the following expressed as milligram per kg. diet: Zn, 100; Mn, 70; I, 0.5; Mo, 5.0; Se, 0.2. 6 The vitamin mixture supplied the following (in units or milligrams per kg. diet): vitamin A, 5300 I.U.; vitamin D3, 400 I.C.U.; thiamine HC1, 3.5; riboflavin, 5.7; Ca pantothenate, 18.5; pyridoxine, HC1, 2.2; niacin, 53; folic acid, 1.5; biotin, .7; B l2 , .018; vitamin K, .8; vitamin E, 22.
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TABLE 1.—Composition of experimental diets used
soya soya + Fe milk milk + Fe, Cu
— —
10.5±2 10.6±.2 9.2±.2 10.1±.2
3 wks.
366±7 330±5
299 ±6 319±7 309±7 302±7
3 wks.
396±6 303 ±9
366±7 368±9 262±12 330±8
9.8 5.5
Trial 1
11.5±.2 10.1 ±.2 6.6±.3 9.2+.2
Trial 2
4 wks.
Age
11.3 4.2
11.8+.4 11.2±.4 5.1±.3 8.9±.4
5 wks.
.05).
Hemoglobin (gms./lOO cc. blood)
352+8 297±9
Trial 1
336±5 332±7 290±9 319+7
Trial 2
Those means with different superscripts were significantly different (P : —data not obtained.
Corn soya Skim milk
Corn Corn Skim Skim
Diet
soya soya + Fe milk milk + Fe, Cu
Corn soya Skim milk
Corn Corn Skim Skim
Diet
RBC counts (x: 10,000) Age 5 wks. 4 wks.
10.5a 4.8 b
10.9s 10.6" 6.9" 9.4C
X
371° 310b
333a 340 s 287" 317c
X
— —
29.4±.4 28.6±.4 27.0±.5 28.5±.5
3 wks.
131 + 1 120+2
120+.9 117+.9 110+1 117+1
3 wks.
TABLE 2.—Hematological response of Cotumix quail fed iron, copper defici
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2104
K. W. WASHBURN AND R. H. LOWE 48
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FIG. 1. Effect of diets on PCV in Coturnix quail. % % Corn-Soya %—0 Corn-Soya + lOOp.p.m. Fe O O Skim Milk O—O Skim Milk + 100 p.p.m. Fe, 5 p.p.m. Cu
RESULTS AND DISCUSSION The packed erythrocyte volume (PCV) response to the skim-milk diet deficient in Fe, Cu was rapid and severe in both trials (Fig. 1) with statistically significant differences being observed one week after initiation. After one week, the PCV of groups in Trial 1 receiving the Fe, Cu deficient diet was 37% compared to 47% for controls receiving the diets adequate in Fe, Cu. A less rapid reduction was observed in Trial 2. After one week
the PCV of the deficient groups in Trial 2 was reduced to 34% compared to 36% for controls receiving the corn-soya turkey starter. The rapid decline in PCV after initiating the deficient diet indicates that Japanese quail are sensitive to deficiencies of this type. The extremely fast growth rate of quail at this age would rapidly dilute the Fe, Cu reserves
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turkey starter (Table 1) containing 250-300 p.p.m. Fe and 10-15 p.p.m. Cu and similar in composition to the corn-soya diet used in Trial 1, but which appeared to give superior growth; 2) the turkey starter to which 100 p.p.m. Fe (as FeS0 4 ) was added; 3) the skim-milk diet used in Trial 1; 4) the skim milk diet to which 100 p.p.m. Fe (as FeS0 4 ) and 5 p.p.m. Cu (as CuS0 4 ) was added. The basal corn-soya diets in each trial were fed until 2 weeks of age, at which time the quail were placed on experimental dietary treatments for a 3-week period. Body weights and packed erythrocyte volume (by the micro-hematocrit method) were obtained at weekly intervals from 2-5 weeks of age for both trials. Feed consumption was measured in Trial 2. Erythrocyte counts, mean cell volumes, hemoglobin concentration and mean cell hemoglobin concentration were determined at intervals shown in Table 2. Erythrocyte counts and mean cell volume were obtained by a model F Coulter Counter and hemoglobin concentration by the acidhematin method of Dennington and Lucas (1955). Two replicates were used for each dietary treatment in each trial with 20 quail used per replicate in Trial 1 and 12 per replicate in Trial 2. Data of two quail from some subsets of Trial 1 and of one quail in Trial 2 were deleted to equalize subclass numbers for statistical analysis by analysis of variance and Duncan's separation of means (Li, 1957).
2105
IRON-COPPER DEFICIENCY IN QUAIL
Further rapid decreases were noted in PCV of quail receiving the skim-milk diet in both trials so that after three weeks the PCV of the Fe, Cu deficient groups in Trial 1 had decreased to 20% compared to 37% in the control group receiving the corn-soya diet. In Trial 2 the PCV of the deficient group decreased to 22% compared to 42% in the control groups receiving the corn-soya diet. Even with this severe reduction in PCV, mortality was slight. In Trial 1, 3/40 quail on Fe, Cu deficient diets and in Trial 2, 1/24 had died by the end of the 5-week experimental period. The addition of 100 p.p.m. Fe to the turkey corn-soya diet in Trial 2 (Fig. 1) did not result in a change in PCV indicating that the amount of Fe in the diet (250-300 p.p.m. Fe) was adequate for maximum hematopoeisis. The addition of Fe and Cu to the skim-milk resulted in a significant elevation of PCV above that of groups receiving the basal skim-milk diet. However, the PCV did not approach the level obtained when the turkey
corn-soya diets were fed. Although the establishment of Fe requirements was not designed into this experiment, this would suggest that the 90-100 p.p.m. Fe requirement for quail suggested by Harland et al. (1973) may not be adequate for normal hematology. Harland et al. (1973) used EDTA extracted soya to produce an Fe deficient diet while in the
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FIG. 2. Effect of diets on growth in Coturnix quail. # % Corn-Soya •----# Corn-Soya +- 100 p.p.m. Fe O O Skim Milk O—O Skim Milk + 100 p.p.m. Fe, 5 p.p.m. Cu
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so that any deficiencies of nutrients necessary for hematopoeisis would quickly be evident. However, the hematology of the quail apparently is not as responsive to chemicals that may have a detrimental effect. Ernst and Ringer (1968) in an evaluation of the effects of pesticide chemicals on erythrocyte of Japanese quail reported that continuous dietary administration of DDT, Zectron or Zytron significantly reduced the PCV and total erythrocyte counts of blood from mature quail but not in immature quail. They concluded that PCV was not a useful criterion of pesticide toxicity since near lethal levels were necessary to cause a significant change in PCV. In contrast to the rapid response observed in quail in this study, Washburn (1969) found that chickens placed on an Fe, Cu deficient diet following feeding of a normal diet for their first two weeks required at least three weeks on the deficient diet to show even a small reduction in PCV.
2106
K. W. WASHBURN AND R. H. LOWE
These results show evidence of the typical hypochromic, microcytic anemia expected from a severe Fe, Cu deficiency. The relative differences in RBC counts, MCV, hemoglo-
TABLE 3.- -Feed conversions (gms. feed/gm.
Age (days) days on diet
bin and MCHC of the groups in Trial 1 and Trial 2 receiving the skim milk diet and the corn-soya diets paralleled differences observed in PCV. The addition of 100 p.p.m. Fe to the corn-soya diet in Trial 2 did not significantly alter the hematological picture. The groups in Trial 2 which were fed the skim-milk diet to which 100 p.p.m. Fe and 5 p.p.m. Cu was added also exhibited a hypochromic, microcytic anemia but the depression in RBC counts, MCV, hemoglobin content and MCHC were much less severe than the groups receiving the skim-milk diet without additional Fe, Cu. The effect of the various diets on growth is presented in Figure 2. The feeding of the Fe, Cu deficient skim-milk diet resulted in approximately 30% reduction in body weight by 5 weeks. This depression in growth may not be primarily attributed to the Fe, Cu deficiency since when quail in Trial 2 were given the skim milk diet with 100 p.p.m. Fe, 5 p.p.m. Cu growth did not approach that of the groups fed the corn-soya diet. Feed conversion ratio (Table 3) was significantly
gain) of Coturnix fed iron-copper deficient or adequate diets (Trial 2) 28-35 14-21
14-35 0-21
3.1 3.2 3.1
21- •28 7- 14 4.0 3.9 4.0
6.6 7.2 6.9
(292) (273) 4.7 a
X
2.7 3.1 2.9
4.1 4.1 4.1
6.5 6.6 6.6
(262) (271) 4.5 a
Skim milk
R, R7 X
3.8 3.8 3.8
5.3 5.0 5.2
9.6 11.1 10.4
(198) (199) 6.5 b
Skim milk + Fe, Cu
R, R, X
3.7 3.9 3.8
5.3 4.4 4.8
5.6 5.4 5.5
(223) (230) 4.7 a
Corn-soya
14-21 0-7 R, R?
X Corn-soya + Fe
R, R?
a
R, = replicate 1; Rj = replicate 2. ( ) = feed consumption. Those means with different superscripts were significantly different (P :
.05).
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present study a skim-milk diet was used. The effects of the various diets on blood measurements other than PCV are presented in Table 2. In Trial 1 after 5 weeks the RBC counts of the groups receiving the skim milk diet were 3.03 million compared to 3.96 million for the groups receiving the corn-soya diet. The MCV was 66 jx-3 compared to 96 (x.3, the hemoglobin content was 4.2 gms. compared to 11.3 gms. and the MCHC was 20.4% compared to 29.1%. Similar changes were observed in Trial 2, the RBC counts of the groups receiving the skim-milk diet were 2.62 million compared to 3.66 million; the MCV was 85JA,.3 compared to 116u.. 3 ; the hemoglobin content was 5.1 gms. compared to 11.8 gms., and the MCHC was 22.9% compared to 27.6%. All these differences were statistically significant as were the differences in mean values for the experimental period.
IRON-COPPER DEFICIENCY IN Q U A I L
1964. Hematology of Coturnix from birth to maturity. Poultry Sci. 43: 1392-1401. Blair, R., M. L. Scott and R. J. Young, 1972. Unidentified factor activities in whole soybeans required for optimum growth of Coturnix. J. Nutr. 102: 1529-1541. Dennington, E. M., and A. M. Lucas, 1955. Blood techniques for chickens. Poultry Sci. 34: 360-368. Ernst, R. A., and R. K. Ringer, 1968. The effect of DDT, Zectran and Zytron on the packed cell volume, total erythrocyte count and mean corpuscular volume of Japanese quail. Poultry Sci. 47: 639-643. Harland, B. F., B. E. Fry, R. M. Jacobs and M. R. S. Fox, 1973. Mineral requirements of young Japanese quail. Federation Proc. 32: 930. Hill, C. H., and C. Matrone, 1961. Studies on copper and iron deficiencies of growing chickens. J. Nutr. 73: 425-431. Jacobs, R. M., M. R. Spivey Foxand M. H. Aldridge, 1969. Changes in plasma proteins associated with the anemia produced by dietary cadmium in Japanese quail. J. Nutr. 99: 119-128. Li,C. R. J., 1957. Introduction to Statistical Inference. Edwards Bros., Inc., Ann Arbor, Michigan. Marks, H. L., 1967. A Japanese quail control population. The Quail Quarterly, 4(1): 2-4. Stino, F. K. R., 1971. Divergent selection for packed erythrocyte volume in the Japanese quail under two environments. Ph.D. Thesis, Univ. of Georgia, Athens. Washburn, K. W., 1969. Hematological response of different stocks of chickens to iron, copper deficient diet. Poultry Sci. 48: 204-209. Wentworth, B. C , 1970. Sterility and reproductive inhibition of Japanese quail induced by mestranol ingestion. Poultry Sci. 49: 1477-1484.
REFERENCES Atwal, O. S., L. Z. McFarland and W. O. WUson,
NEWS AND NOTES (Continued from page 2085) received a Ph.D. from Louisiana State University, majoring in poultry nutrition. He was winner of the Charles Upp Award for outstanding graduate work in Poultry Science at L.S.U., and will continue to serve as Valmac's chief nutritionist.
Before joining Valmac, he was Poultry Nutritionist with Pillsbury Farms and Manager of the poultry research farm at L.S.U. Valmac Industries is one of the nation's large poultry processors with four plants in Arkansas at Russellville, Dardanelle, Pine Bluff and Waldrop.
(Continued on page 2136)
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increased in the groups fed the milk diet without supplemental iron, but conversion was not different in the groups receiving the skim-milk diet supplemented with Fe and Cu from groups receiving the corn-soya diet. The results of Blair et al. (1972) suggest that quail chicks fed an amino acid deficient diet require certain peptides for maximizing growth. When isolated soy protein was substituted for a portion of the amino acids in a basal amino acid diet, growth was increased by 35% and by 54% when heated ground soybeans were used. This apparent requirement for peptides obtainable from soy protein is a logical explanation for the growth depression of the quail given the semi-purified skim-milk diet. A number of factors have been shown to affect the hematology of quail; however, there is no evidence that rate of growth per se has any relationship. Wentworth (1970), in a study of the effects of mestranol ingestion on sterility and reproduction in Japanese quail, reported that both castration and mestranol ingestion significantly depressed PCV in sexually mature males. Jacobs et al. (1969) produced an anemia in Japanese quail by use of dietary cadmium. This anemia was associated with increased levels of plasma transferrins.
2107