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Effects of Diet Composition on Vanadium Toxicity in Laying Hens1
Effects of Diet Composition on Vanadium Toxicity in Laying Hens1 L. E. OUSTERHOUT and L. R. BERG 2 Department of Animal Sciences, Western Washington Research and Extension Center, Washington State University, Puyallup, Washington 983 71 (Received for publication September 2, 1980)
1981 Poultry Science 60:1152-1159 INTRODUCTION Berg et al. ( 1 9 6 3 ) e x t e n d e d their vanadium toxicity studies from growing chicks t o laying hens and found t h a t vanadium a d d e d t o t h e diet caused a significant d e t e r i o r a t i o n in a l b u m e n quality as measured by Haugh units. Levels between 0 and 6 0 p p m depressed Haugh units in a linear relationship. Levels of 3 0 p p m and above depressed egg p r o d u c t i o n with 1 0 0 p p m almost stopping lay after 3 weeks. Levels of 4 0 p p m and above depressed hatchability and all levels of vanadium caused a loss in b o d y weight. T h e depression in Haugh units was a p p a r e n t after 5 days. By 5 weeks, depressions were as great as 15 units. Effects were similar w h e n vanadium was a d d e d as N H 4 V O 3 or as a p h o s p h a t e c o n t a m i n a n t in t h e form of V 2 O 5 . However, Hafez and Kratzer ( 1 9 7 6 b ) f o u n d t h a t 2 0 0 p p m added vanadium reduced rate of
'Scientific Paper No. 5725. College of Agriculture Research Center, Washington State University, Projects 0500 and 1748. 2 Deceased.
lay b u t 1 0 0 p p m had n o effect. T h e y did n o t r e p o r t on a l b u m e n quality b u t f o u n d t h a t vanadium h a d n o significant effect on egg weight, egg shell thickness, or egg y o l k cholesterol. Laying C o t u r n i x quail fed t h e same diets were n o t affected by 2 0 0 p p m vanadium and showed only a nonsignificant drop in egg p r o d u c t i o n at 3 0 0 p p m . J e n s e n and Maurice ( 1 9 8 0 ) f o u n d t h a t 5 t o 10 p p m a d d e d chrom i u m was partially effective in counteracting t h e d e t e r i o r a t i o n in a l b u m e n quality caused b y 10 or 2 0 p p m a d d e d v a n a d i u m . O t h e r dietary changes h a d been r e p o r t e d t o alleviate t h e g r o w t h depression caused by vanadium in chicks. Berg ( 1 9 6 6 ) reported t h a t certain sources of protein and c a r b o h y d r a t e , dietary levels of magnesium and potassium, and t h e protein level of t h e diet modified t h e g r o w t h depression and a m o u n t of m o r t a l i t y caused b y 2 0 p p m a d d e d v a n a d i u m . Hill ( 1 9 7 9 b ) reported an alleviation of t h e g r o w t h depressing effect of 3 to 12 p p m vanadium w h e n t h e p r o t e i n level of a soy-glucose basal was increased from 10 to 2 0 o r 30%. Berg and Lawrence ( 1 9 7 1 ) r e p o r t e d t h a t additions of 5% c o t t o n s e e d meal, 5% d e h y d r a t e d grass, or . 2 5 % ascorbic
1152
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ABSTRACT Vanadium added to laying rations as NH4 V 0 3 , VOCl2 or VOS0 4 at levels of 20 to 80 ppm resulted in a rapid and substantial reduction in albumen quality as measured by Haugh units. Dietary vanadium also resulted in reduced egg production, egg weight, body weight, feed consumption, and poorer shell quality as measured by specific gravity. Ascorbic acid at .4 to .5% effectively protected the hen from the reduction in albumen quality, egg production, and body weight for up to 40 ppm vanadium, but not the reduction of egg weight Replacement of soybean meal by 20% dietary cottonseed meal also protected the hen from the reduction in albumen quality, egg production, and body weight for up to 40 ppm vanadium. Added at levels of 4 to 8 times the molecular concentration of vanadium, EDTA had no consistent effect on vanadium toxicity. Dehydrated grass, at levels of 6 to 12%, maintained egg production but had no effects on the reduction in albumen quality caused by 40 ppm vanadium. Replacement of soybean meal with herring fish meal and part of the grain with sucrose intensified the depression of albumen quality, egg production, and loss of body weight caused by added vanadium. Neither varying dietary protein levels from 12 to 25% using soybean meal nor the addition of 20 ppm chromium had any effect on the toxicity of added vanadium. It appears that vanadium expresses its toxicity in laying hens by several routes since the protective effects of different dietary changes and additives differentially affected the loss of albumen quality, egg production, body weight, and egg weight. (Key words: vanadium, Haugh units, albumen quality, chromium, cottonseed meal, dehydrated grass, EDTA, ascorbic acid, egg production, egg size, shell quality)
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5.80
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5.65
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3 8.75
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2.5 7.75
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6.00
5.88
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60.274
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72.445
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17.65
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Experiment and diet
25 17.1 25
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studies
1
7
4
Supplied as in 2 except: 2.2 mg calcium pantothenate.
Supplied as in 2 except: 2.2 mg calcium pantothenate, 0 niacin, choline chloride, and vitamin B n .
Supplied as in 1 except: 3300 IU vitamin A, 770 ICU vitamin D, 4.4 mg calcium pantothenate, 8.8 mg niacin, 220
Supplied as in 1 except: 0 ethoxyquin.
'Supplied as in 1 except: 2.2 mg menadione sodium bisulfite complex.
6
5 Supplied (per kg diet): 7900 IU vitamin A, 2000 ICU vitamin D, 2.25 IU vitamin E, 2.3 mg menadione sodium d-pantothenate, 29 mg niacin, 280 mg choline chloride, .1 mg vitamin B 1 2 , 125 mg butylated hydroxytoluene, 60 iron (carbonate), 2 mg copper (oxide), 1.2 mg iodine (iodate), .2 mg cobalt (carbonate).
4
3
2
1 Supplied (per kg of diet): 4400 IU vitamin A, 1100 ICU vitamin D, 3.3 mg riboflavin, 66 mg calcium pantothe 125 mg ethoxyquin.
Ground corn Ground sorghum Ground barley Sucrose Dehydrated grass Soybean meal (50%) Cottonseed meal (44%) Meat and bone meal Herring fish meal Peruvian fish meal Tri-calcium phosphate Defluorinated phosphate Ground limestone Tallow Salt DL-methione Vitamin and mineral mix
2B
2A
1
TABLE 1. Basal diets used in vanadium toxicity
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OUSTERHOUT AND BERG
The experiments reported here were designed to clarify the effects of vanadium on the performance of laying hens and to study the effects of dietary changes and additions on albumen quality, egg production, egg weight, body weight and shell quality.
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METHODS AND MATERIALS The experimental diets (Table 1) were fed to groups of individually caged White Leghorn hens kept in a windowless fan-ventilated house and given 14 hr of artificial light daily. Except where noted, all hens were fed either the basal
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acid to a fish meal-sucrose basal diet markedly reduced or eliminated the growth depression caused by 20 ppm added vanadium. Other protein sources had little or no effect on vanadium toxicity. Neither gland-free cottonseed meal nor gossypol protected the chicks from vanadium toxicity. Hill (1979a) reported" .1 or .2% ascorbic acid effectively prevented the growth depression caused by 20 ppm added vanadium. Hafez and Kratzer (1976a) reported an increased vanadium toxicity with a semipurified (isolated soy protein-corn starch) diet compared to one containing natural feed ingredients with 100 ppm added vanadium causing 100% mortality of chicks fed the semipurified diet but no mortality and only 17.5% growth depression with the natural diet. The natural diet contained 5.25% cottonseed meal, 5% dehydrated alfalfa meal, and other ingredients that might be expected, on the basis of the work of Berg and Lawrence (1971), to reduce vanadium toxicity. With a different natural diet (no cottonseed meal), they reported a partial alleviation of the growth depression and mortality caused by 5 to 200 ppm added vanadium by the addition of disodium ethylenediaminetetraacetate (EDTA) equivalent to 6 times the molar concentration of 50 ppm vanadium. Addition of 10% lactose to the diet increased the growth depression and caused 80% mortality with 100 ppm vanadium. With a third natural diet (1.5% cottonseed meal and inclusion of several other high fiber ingredients), addition of 1000 ppm chromium reduced the mortality and growth depression caused by 100 ppm vanadium. Hathcock et al. (1964) had previously reported that EDTA at 2 or 6 times the molar concentration of vanadium practically eliminated the toxicity symptoms caused by 25 ppm vanadium added to a skim milk-glucose basal diet.
ormance
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VANADIUM TOXICITY IN LAYING HENS
1155
TABLE 3. Effects of dietary protein level and EDTA1 on albumen quality deterioration caused by vanadium toxicity in laying bens (Experiment 2) Treatment Protein (%)
Haugh units
Vanadium (ppm)
EDTA
12 4X1
40 40 Mean
4X
40 40 Mean
4X
40 40 Mean
4X
15
20
25
4 wk
Change
72.8 78.5 71.3 74.2 78.2 74.1 75.6 76.0 78.5 72.9 75.7 75.7 79.8 75.2 72.3 75.8
74.2 68.8 68.2 70.4 76.3 65.6 66.9 69.6 80.0 62.0 70.0 70.7 79.5 63.1 71.4 71.3
+1.4 -9.7 -3.1 -3.8 -1.9 -8.5 -8.7 -6.4 +1.5 -10.9 -5.7 -5.0 -.3 -12.1 -.9 -^.5
77.3 75.2 73.7
77.5 64.9 69.1
+0.2 a -10.3C -4.6b
Averages 40 40
4X
a ' ' c Groups of means followed by a different superscript are significantly different (P<.01). Protein level had no significant effect. 1
EDTA was added as disodium ethylenediaminetetraacetate at 4 times the molar concentration of vanadium.
diet o r o n e very similar for at least t w o w e e k s prior t o t h e 4 week e x p e r i m e n t a l period. In all trials e x c e p t E x p e r i m e n t 5C, t h e vanadium was a d d e d as a m m o n i u m vanadate ( N H 4 V O 3 ) . Eggs were gathered daily and t h e rate of lay was calculated as a p e r c e n t hen day p r o d u c t i o n for
t h e period given. Haugh units, egg weights, and specific gravity were m e a s u r e d after holding t h e eggs overnight in a refrigerated egg r o o m a n d were normally d e t e r m i n e d o n all eggs laid on t h r e e consecutive days at t h e t i m e indicated.
TABLE 4. Effects of type of diet and EDTA* on vanadium toxicity in laying hens (Experiment 3) Haugh units
Treatment Basal
Vanadium
EDTA
Pretest
1 wk
4 wk
Change
84.5 90.8 88.1 86.1 91.4 84.5 86.4 89.4
81.7 83.2 80.9 79.5 82.0 74.0 84.0 79.6
80.9 74.7 70.2 80.0 73.2 69.2 81.5 67.3
-3.6^ -16.lt>c -17.9cd -6.1» -18.2= d -15.3DC -4.9a -22.1d
(%)
(ppm) Mixed 40 40
4XX
40 40
4X
Corn-fish Corn-sucrose-fish 40 a
Rate of lay
80a 77a 70* 80a 71a 83a 78a 50b
Body weight change (g) +96 a +64ab +60ab +86 a -23 b c +50ab +118 a +105 a
' ' c ' Means in a column not followed by a common superscript are significantly different (P<.05).
1
EDTA was added as disodium ethylenediaminetetraacetate at 4 times the molar concentration of vanadium.
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40 40 Mean
Pretest
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OUSTERHOUT AND BERG
TABLE 5. The effects of dietary ascorbic acid (AA) on vanadium toxicity in laying hens (Experiment 4)
0% AA
0 40 80
75.8bc 71.lb 63.la
.2% AA
b
73.3 72.3 b
Egg weight (g)
Rate of lay (%)
Haugh units (ppm)
.4% AA
0% AA
.2% AA
.4% AA
0% AA
79.7C 76.3bc 72.2b
82.5b 77.lb 47.1*
84.3 b 85.7 b
84.3 b 88.6 b 85.7 b
64.6 a 60.1= 60.5 C
.2% AA
.4% AA 62.8 b 60.4 C 61.0 C
C
60.8 61.6bc
a,b,c Means for a parameter not followed by a common superscript are significantly different (P<.05).
weeks of age were assigned to each treatment. Diet 5 A (Table 1) was the basal used for Experiments 5A and 5C and was the soy basal for Experiment 5B. Diets 5B and 5C were the 10 and 20% cottonseed meal basals for Experiment 5B. Experiment 6. One group of 10 hens 50 weeks of age was assigned to each treatment. Chromium was added as OCI3 • 6 H 2 0 . Data are for the 6 days following a 4 day adjustment period on the experimental diets. Experiment 7. Two groups of 10 hens 35 weeks of age were assigned to each treatment except 4 groups were fed the control diet. The dehydrated grass was added to the basal diet at the expense of ground corn. Where statistical differences in performance are indicated, significance was determined using Duncan's new multiple range test following an analysis of variance that indicated a factor to have significant effects. Where pretest data are given and more than a single pen is assigned a treatment, the analysis was made on the change in value of the factor measured for each
TABLE 6. The effects of ascorbic acid and EDTA1 on vanadium toxicity in laying hens (Experiment 5A) Treatment Vanadium (ppm) 0 40 40 40 40 40
Haugh units
Ascorbic acid
Rate lay
Body weight change
(%)
EDTA
Pretest
4 wk
Change
(%)
(g)
0 0
0 0 0 6X 1 8X 6X
78.6 81.0 80.5 80.7 80.4 81.6
76.5 64.2 78.7 69.1 68.6 81.5
-2.ia -16.8b -1.8a -11.6b -11.8b -.ia
65a 62a 75a 73a 70 a 77a
+17a -15a +4 a -29a _9a +6a
.5 0 0 .5
a' bMeans in a column followed by a common superscript are not significantly different (P>.05). 1 EDTA was added as disodium ethylenediaminetetraacetate at 6 and 8 times the molar concentration of vanadium.
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Specific differences in the various experiments are: Experiment 1, Two groups of 10 hens 50 weeks of age were assigned to each treatment with the experimental diets being fed for 7 weeks. Experiment 2. One group of 10 hens 60 weeks of age was assigned to each treatment. Ration 2A (Table 1) was calculated to be 12%; 2B, 15%; 2C, 20%; and 2D, 25% protein. Experiment 3. Two groups of 10 hens 50 weeks of age were assigned to each treatment. Ration 3 A (Table 1) was the mixed grain basal, 3B the corn-fish basal, and 3C the corn-sucrosefish basal diet. Haugh units were determined on two consecutive eggs per hen per treatment prior to feeding the experimental diets and again after 4 weeks on trial. Experiment 4. One group of 10 hens 48 weeks of age was assigned to each treatment. Haugh units and rate of lay were determined using all eggs laid during the second week of the trial. Experiment 5. Two groups of 10 hens 40
VANADIUM TOXICITY IN LAYING HENS
1157
TABLE 7. The effects of cottonseed meal (CSM) on the vanadium toxicity in laying hens (Experiment 5B) Haugh units
Treatment Vanadium (ppm) Pretest
Basal Soy
40 10% CSM 40 20% CSM 40
78.6 81.0 80.2 81.4 80.4 82.6
4 wk
Change
Rate of lay (%)
Body weight change (g)
76.5 64.2 77.5 71.4 80.4 76.2
-2.1* -16.8b -2.7* -10.0ab 0» -4.4a
65bc 62= 74ab 7jab 77a 77a
+17*b -ISa +60 b +60b +72b +5lb
pen during the test. Where only a single pen is assigned a treatment, the analysis was made using the mean of data for each individual hen for the period indicated. RESULTS AND DISCUSSION Tabulated results for each experiment are given in Tables 2 through 10. In these experiments and in others not reported here, the addition of vanadium at levels of 20 ppm and above quickly (within 3 to 4 days) caused a substantial deterioration in albumen quality. The albumen quality continued to decline for at least 7 weeks. Levels of 80 ppm and above caused near cessation of egg production and considerable molting. Vanadium at 40 ppm usually reduced egg production. Removal of vanadium from the diets allowed albumen quality and egg production to return to normal, the time required depending on the length of the feeding period and the level at which vanadium was fed. Only slight differences in effects were found when the different vana-
dium compounds were compared (Table 8). VOCI2 appeared to be less toxic than VOSO4 orNH4V03. Dietary vanadium at 40 ppm fed for 4 weeks or more also significantly reduced feed intake, body weight, egg weight, and shell quality (Table 2). The effect on egg size and shell quality is contrary to other reports (Berg et al., 1963; Hafez and Kratzer, 1976b). The unique effect is the rapid deterioration in albumen viscosity as measured by Haugh units. The degree of depression of Haugh units and the other effects caused by vanadium depends upon the type of diet and upon certain dietary additives. The use of herring fish meal rather than soybean meal as the supplemental protein source coupled with partial replacement of the grain with sucrose intensified the depression of Haugh units, body weight, and rate of lay (Table 4). This agrees with the effects on growing chicks (Berg, 1966; Hafez and Kratzer, 1976a). Dehydrated grass added at levels from 6 to 12% of the diet prevented the depression of egg production but had little effect on the
TABLE 8. Effect of different vanadium compounds on the performance of laying hens (Experiment 5C) Vanadium Source Control NH„VO., VOCl2
voso4
Haugh units
leve' (ppm)
Pretest
2wk
4 wk
Change
Rate of lay (%)
Body weight change (g)
20 40 20 40 20 40
77.6 77.4 79.6 76.5 78.6 78.4 79.0
78.5 69.6 68.2 71.1 69.7 71.7 68.5
78.7 67.3 65.7 69.9 68.8 72.2 68.3
+l.la -10. l b -13.9b -6.6ab -9.8b -6.2*b -10.7b
74a 68a 57b 69a 71a 70 a 65 a b
+20 a +iab -46b +18 a 0ab +32 a +2ab
Means in a column not followed by a common superscript are significantly different (P<.05).
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' ' Means in a column followed by a common superscript are not significantly different (P>.05).
1158
OUSTERHOUT AND BERG
TABLE 9. Effect of dietary chromium on albumen quality deterioration caused by dietary vanadium in laying hens (Experiment 6) Chromium 0
20 Haugh units
Vanadium
/
81.3 b c go.iabc 76.5 a 77.7ab
^ 83 .OC 79.3abc 76.5a 76.3 a
' ' Means for a parameter not followed by a common superscript are significantly different (P<.05).
maintenance of albumen quality caused by 20 to 40 ppm vanadium (Table 10). In chicks, 5% dehydrated grass almost prevented the growth depression (Berg and Lawrence, 1971). It appears that vanadium acts through different metabolic routes in its effects on albumen quality and on egg production. Dietary protein levels between 12 and 25% obtained by increasing the soybean meal in the diet had no observable effect on the reduction
TABLE 10. Effects of dehydrated grass on vanadium toxicity in laying hens (Experiment 7) Haugh units
Treatment 1
Grass (%) 0
3 6 12 Averages 0 3 6 12
Vanadium (ppm)
Pretest
4 wk
Change
Rate of lay ( 2 - 4 wk) (%)
0 20 40 20 40 20 40 20 40
90.0 88.4 89.5 89.8 90.0 90.2 91.4 91.0 89.8
82.6 74.1 74.2 74.4 68.0 77.6 72.8 78.4 73.4
-7.4a -14.3b -14.3b -15.4b -22.0C -12.6ab -18.6bc -12.6ab -16.4bc
76ab 5 gbcd 40d 76ab 50cd 84a 70abc 7^abc 76ab
20-40 20-40 20-40 20-40 0 20 40
89.0 89.9 90.8 90.4 90.0 89.8 90.2
74.7 71.2 75.2 75.9 82.6 76.1 72.3
-14.3 -17.7 -15.6 -14.5 -7.4a -13.7b -17.9C
49a 63b 77b 73b 76b 70b 59 a
' ' ' Means in a column or group of averages not followed by a common superscript are significandy different (P<.05). 1 Dehydrated grass was added at the expense of ground corn.
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(ppm) 0 20 40 80
in albumen quality caused by 40 ppm vanadium (Table 3). Hill (1979b) had reported a protective effect for higher protein levels with chicks. However, replacing all or at least half of the soybean and meat and bone meal with cottonseed meal almost prevented Haugh unit depression caused by 40 ppm vanadium and increased egg production and body weight over that of the controls (Table 7). Effects of lower levels of cottonseed meal, similar to those that prevented growth depression with chicks (Berg and Lawrence, 1971), had no discernible effects and are not reported here. Because EDTA had been reported by Hathcock et al. (1964) and Hafez and Kratzer (1976a) to alleviate vanadium toxicity in chicks, it was tested singly and in combination with other factors to determine its effects on laying hens. With 20 or 25% protein diets, it protected against a loss of albumen quality. It had no significant effect on egg production or body weight (Tables 3, 4, and 6). Chromium added to the diets at 20 ppm had no detectible effect in preventing the albumen quality deterioration caused by 20 to 80 ppm vanadium (Table 9); this is contrary to the report of Jensen and Maurice (1980). Perhaps the protective effect of chromium takes longer to develop than the 10 days of this trial. Their
VANADIUM TOXICITY IN LAYING HENS trials ran 4 to 6 weeks. Regression coefficients for Haugh units and vanadium levels were: No chromium: Haugh units = 80.6 - .048 x V (ppm), r = - . 7 4 8 20 ppm chromium: Haugh units = 81.6 - .080 X V (ppm), r = - . 8 6 8
the diet and the age and strain of the hens. REFERENCES Berg, L. R., 1966. Effect of diet composition on vanadium toxicity for the chick. Poultry Sci. 45:1346-1352. Berg, L. R., G. E. Bearse, and L. H. Merrill, 1963. Vanadium toxicity in laying hens. Poultry Sci. 42:1407-1411. Berg, L. R., and W. W. Lawrence, 1971. Cottonseed meal, dehydrated grass and ascorbic acid as dietary factors preventing toxicity of vanadium for the chick. Poultry Sci. 50:13991404. Chen, A. A-T., and C. F. Nockels, 1973. The effects of dietary vitamin C, protein, strain and age on egg quality production and serum and albumen protein of chickens. Poultry Sci. 52:1862—1867. Hafez, Y.S.M., and F. H. Kratzer, 1976a. The effect of diet on the toxicity of vanadium. Poultry Sci. 55:918-922. Hafez, Y.S.M., and F. H. Kratzer, 1976b. The effect of pharmacological levels of dietary vanadium on the egg production, shell thickness and egg yolk cholesterol in laying hens and coturnix. Poultry Sci. 55:923-926. Hathcock, J. N., C. H. Hill, and G. Matrone, 1964. Vanadium toxicity and distribution in chicks and rats. J. Nutr. 82:106-110. Hill, C. H., 1979a. Studies on the ameliorating effect of ascorbic acid on mineral toxicities in the chick. J. Nutr. 109:84-90. Hill, C. H., 1979b. The effect of dietary protein levels on mineral toxicity in the chick. J. Nutr. 109: 501-507. Jensen, L. S., and D. V. Maurice, 1980. Dietary chromium and interior egg quality. Poultry Sci. 59:341-346.
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Although the coefficients were not significantly different, there was an indication that the chromium addition improved the Haugh units in the basal diet, but the improvement was quickly lost when vanadium was added. This trial was too short for the expected loss of egg production normally caused by 80 ppm vanadium to be significant. The addition of ascorbic acid at .4 or .5% completely prevented the depression of albumen quality and egg production caused by up to 40 ppm vanadium (Tables 5 and 6). However, dietary ascorbic acid did not prevent the egg weight depression caused by vanadium and appeared to depress the egg weight from the diet without added vanadium. Berg and Lawrence (1971) and Hill (1979a) have reported that .1 to .25% ascorbic acid prevented vanadium toxicity with chicks caused by 20 ppm vanadium. Chen and Nockels (1973) have reported that the effect of ascorbic acid on egg production, egg weight, and Haugh units depends upon the protein content of
1159
1981 Poultry Science 60:1152-1159 INTRODUCTION Berg et al. ( 1 9 6 3 ) e x t e n d e d their vanadium toxicity studies from growing chicks t o laying hens and found t h a t vanadium a d d e d t o t h e diet caused a significant d e t e r i o r a t i o n in a l b u m e n quality as measured by Haugh units. Levels between 0 and 6 0 p p m depressed Haugh units in a linear relationship. Levels of 3 0 p p m and above depressed egg p r o d u c t i o n with 1 0 0 p p m almost stopping lay after 3 weeks. Levels of 4 0 p p m and above depressed hatchability and all levels of vanadium caused a loss in b o d y weight. T h e depression in Haugh units was a p p a r e n t after 5 days. By 5 weeks, depressions were as great as 15 units. Effects were similar w h e n vanadium was a d d e d as N H 4 V O 3 or as a p h o s p h a t e c o n t a m i n a n t in t h e form of V 2 O 5 . However, Hafez and Kratzer ( 1 9 7 6 b ) f o u n d t h a t 2 0 0 p p m added vanadium reduced rate of
'Scientific Paper No. 5725. College of Agriculture Research Center, Washington State University, Projects 0500 and 1748. 2 Deceased.
lay b u t 1 0 0 p p m had n o effect. T h e y did n o t r e p o r t on a l b u m e n quality b u t f o u n d t h a t vanadium h a d n o significant effect on egg weight, egg shell thickness, or egg y o l k cholesterol. Laying C o t u r n i x quail fed t h e same diets were n o t affected by 2 0 0 p p m vanadium and showed only a nonsignificant drop in egg p r o d u c t i o n at 3 0 0 p p m . J e n s e n and Maurice ( 1 9 8 0 ) f o u n d t h a t 5 t o 10 p p m a d d e d chrom i u m was partially effective in counteracting t h e d e t e r i o r a t i o n in a l b u m e n quality caused b y 10 or 2 0 p p m a d d e d v a n a d i u m . O t h e r dietary changes h a d been r e p o r t e d t o alleviate t h e g r o w t h depression caused by vanadium in chicks. Berg ( 1 9 6 6 ) reported t h a t certain sources of protein and c a r b o h y d r a t e , dietary levels of magnesium and potassium, and t h e protein level of t h e diet modified t h e g r o w t h depression and a m o u n t of m o r t a l i t y caused b y 2 0 p p m a d d e d v a n a d i u m . Hill ( 1 9 7 9 b ) reported an alleviation of t h e g r o w t h depressing effect of 3 to 12 p p m vanadium w h e n t h e p r o t e i n level of a soy-glucose basal was increased from 10 to 2 0 o r 30%. Berg and Lawrence ( 1 9 7 1 ) r e p o r t e d t h a t additions of 5% c o t t o n s e e d meal, 5% d e h y d r a t e d grass, or . 2 5 % ascorbic
1152
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ABSTRACT Vanadium added to laying rations as NH4 V 0 3 , VOCl2 or VOS0 4 at levels of 20 to 80 ppm resulted in a rapid and substantial reduction in albumen quality as measured by Haugh units. Dietary vanadium also resulted in reduced egg production, egg weight, body weight, feed consumption, and poorer shell quality as measured by specific gravity. Ascorbic acid at .4 to .5% effectively protected the hen from the reduction in albumen quality, egg production, and body weight for up to 40 ppm vanadium, but not the reduction of egg weight Replacement of soybean meal by 20% dietary cottonseed meal also protected the hen from the reduction in albumen quality, egg production, and body weight for up to 40 ppm vanadium. Added at levels of 4 to 8 times the molecular concentration of vanadium, EDTA had no consistent effect on vanadium toxicity. Dehydrated grass, at levels of 6 to 12%, maintained egg production but had no effects on the reduction in albumen quality caused by 40 ppm vanadium. Replacement of soybean meal with herring fish meal and part of the grain with sucrose intensified the depression of albumen quality, egg production, and loss of body weight caused by added vanadium. Neither varying dietary protein levels from 12 to 25% using soybean meal nor the addition of 20 ppm chromium had any effect on the toxicity of added vanadium. It appears that vanadium expresses its toxicity in laying hens by several routes since the protective effects of different dietary changes and additives differentially affected the loss of albumen quality, egg production, body weight, and egg weight. (Key words: vanadium, Haugh units, albumen quality, chromium, cottonseed meal, dehydrated grass, EDTA, ascorbic acid, egg production, egg size, shell quality)
2.00
5.80
.3
.25'
5.65
.3
.25'
3 8.75
2.5 .35
5
2.5 7.75
.3 .046 .25'
6.00
5.88
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1.63
3 28.5
60.274
1.86
3 16.25
72.445
79.9
75.7
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48.281
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1.08
17.65
35 3
37.47
3C
Experiment and diet
25 17.1 25
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studies
1
7
4
Supplied as in 2 except: 2.2 mg calcium pantothenate.
Supplied as in 2 except: 2.2 mg calcium pantothenate, 0 niacin, choline chloride, and vitamin B n .
Supplied as in 1 except: 3300 IU vitamin A, 770 ICU vitamin D, 4.4 mg calcium pantothenate, 8.8 mg niacin, 220
Supplied as in 1 except: 0 ethoxyquin.
'Supplied as in 1 except: 2.2 mg menadione sodium bisulfite complex.
6
5 Supplied (per kg diet): 7900 IU vitamin A, 2000 ICU vitamin D, 2.25 IU vitamin E, 2.3 mg menadione sodium d-pantothenate, 29 mg niacin, 280 mg choline chloride, .1 mg vitamin B 1 2 , 125 mg butylated hydroxytoluene, 60 iron (carbonate), 2 mg copper (oxide), 1.2 mg iodine (iodate), .2 mg cobalt (carbonate).
4
3
2
1 Supplied (per kg of diet): 4400 IU vitamin A, 1100 ICU vitamin D, 3.3 mg riboflavin, 66 mg calcium pantothe 125 mg ethoxyquin.
Ground corn Ground sorghum Ground barley Sucrose Dehydrated grass Soybean meal (50%) Cottonseed meal (44%) Meat and bone meal Herring fish meal Peruvian fish meal Tri-calcium phosphate Defluorinated phosphate Ground limestone Tallow Salt DL-methione Vitamin and mineral mix
2B
2A
1
TABLE 1. Basal diets used in vanadium toxicity
ttp://ps.oxfordjournals.org/ at Universitat Autònoma de Barcelona on October 29, 2014
OUSTERHOUT AND BERG
The experiments reported here were designed to clarify the effects of vanadium on the performance of laying hens and to study the effects of dietary changes and additions on albumen quality, egg production, egg weight, body weight and shell quality.
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METHODS AND MATERIALS The experimental diets (Table 1) were fed to groups of individually caged White Leghorn hens kept in a windowless fan-ventilated house and given 14 hr of artificial light daily. Except where noted, all hens were fed either the basal
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acid to a fish meal-sucrose basal diet markedly reduced or eliminated the growth depression caused by 20 ppm added vanadium. Other protein sources had little or no effect on vanadium toxicity. Neither gland-free cottonseed meal nor gossypol protected the chicks from vanadium toxicity. Hill (1979a) reported" .1 or .2% ascorbic acid effectively prevented the growth depression caused by 20 ppm added vanadium. Hafez and Kratzer (1976a) reported an increased vanadium toxicity with a semipurified (isolated soy protein-corn starch) diet compared to one containing natural feed ingredients with 100 ppm added vanadium causing 100% mortality of chicks fed the semipurified diet but no mortality and only 17.5% growth depression with the natural diet. The natural diet contained 5.25% cottonseed meal, 5% dehydrated alfalfa meal, and other ingredients that might be expected, on the basis of the work of Berg and Lawrence (1971), to reduce vanadium toxicity. With a different natural diet (no cottonseed meal), they reported a partial alleviation of the growth depression and mortality caused by 5 to 200 ppm added vanadium by the addition of disodium ethylenediaminetetraacetate (EDTA) equivalent to 6 times the molar concentration of 50 ppm vanadium. Addition of 10% lactose to the diet increased the growth depression and caused 80% mortality with 100 ppm vanadium. With a third natural diet (1.5% cottonseed meal and inclusion of several other high fiber ingredients), addition of 1000 ppm chromium reduced the mortality and growth depression caused by 100 ppm vanadium. Hathcock et al. (1964) had previously reported that EDTA at 2 or 6 times the molar concentration of vanadium practically eliminated the toxicity symptoms caused by 25 ppm vanadium added to a skim milk-glucose basal diet.
ormance
1154
VANADIUM TOXICITY IN LAYING HENS
1155
TABLE 3. Effects of dietary protein level and EDTA1 on albumen quality deterioration caused by vanadium toxicity in laying bens (Experiment 2) Treatment Protein (%)
Haugh units
Vanadium (ppm)
EDTA
12 4X1
40 40 Mean
4X
40 40 Mean
4X
40 40 Mean
4X
15
20
25
4 wk
Change
72.8 78.5 71.3 74.2 78.2 74.1 75.6 76.0 78.5 72.9 75.7 75.7 79.8 75.2 72.3 75.8
74.2 68.8 68.2 70.4 76.3 65.6 66.9 69.6 80.0 62.0 70.0 70.7 79.5 63.1 71.4 71.3
+1.4 -9.7 -3.1 -3.8 -1.9 -8.5 -8.7 -6.4 +1.5 -10.9 -5.7 -5.0 -.3 -12.1 -.9 -^.5
77.3 75.2 73.7
77.5 64.9 69.1
+0.2 a -10.3C -4.6b
Averages 40 40
4X
a ' ' c Groups of means followed by a different superscript are significantly different (P<.01). Protein level had no significant effect. 1
EDTA was added as disodium ethylenediaminetetraacetate at 4 times the molar concentration of vanadium.
diet o r o n e very similar for at least t w o w e e k s prior t o t h e 4 week e x p e r i m e n t a l period. In all trials e x c e p t E x p e r i m e n t 5C, t h e vanadium was a d d e d as a m m o n i u m vanadate ( N H 4 V O 3 ) . Eggs were gathered daily and t h e rate of lay was calculated as a p e r c e n t hen day p r o d u c t i o n for
t h e period given. Haugh units, egg weights, and specific gravity were m e a s u r e d after holding t h e eggs overnight in a refrigerated egg r o o m a n d were normally d e t e r m i n e d o n all eggs laid on t h r e e consecutive days at t h e t i m e indicated.
TABLE 4. Effects of type of diet and EDTA* on vanadium toxicity in laying hens (Experiment 3) Haugh units
Treatment Basal
Vanadium
EDTA
Pretest
1 wk
4 wk
Change
84.5 90.8 88.1 86.1 91.4 84.5 86.4 89.4
81.7 83.2 80.9 79.5 82.0 74.0 84.0 79.6
80.9 74.7 70.2 80.0 73.2 69.2 81.5 67.3
-3.6^ -16.lt>c -17.9cd -6.1» -18.2= d -15.3DC -4.9a -22.1d
(%)
(ppm) Mixed 40 40
4XX
40 40
4X
Corn-fish Corn-sucrose-fish 40 a
Rate of lay
80a 77a 70* 80a 71a 83a 78a 50b
Body weight change (g) +96 a +64ab +60ab +86 a -23 b c +50ab +118 a +105 a
' ' c ' Means in a column not followed by a common superscript are significantly different (P<.05).
1
EDTA was added as disodium ethylenediaminetetraacetate at 4 times the molar concentration of vanadium.
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40 40 Mean
Pretest
1156
OUSTERHOUT AND BERG
TABLE 5. The effects of dietary ascorbic acid (AA) on vanadium toxicity in laying hens (Experiment 4)
0% AA
0 40 80
75.8bc 71.lb 63.la
.2% AA
b
73.3 72.3 b
Egg weight (g)
Rate of lay (%)
Haugh units (ppm)
.4% AA
0% AA
.2% AA
.4% AA
0% AA
79.7C 76.3bc 72.2b
82.5b 77.lb 47.1*
84.3 b 85.7 b
84.3 b 88.6 b 85.7 b
64.6 a 60.1= 60.5 C
.2% AA
.4% AA 62.8 b 60.4 C 61.0 C
C
60.8 61.6bc
a,b,c Means for a parameter not followed by a common superscript are significantly different (P<.05).
weeks of age were assigned to each treatment. Diet 5 A (Table 1) was the basal used for Experiments 5A and 5C and was the soy basal for Experiment 5B. Diets 5B and 5C were the 10 and 20% cottonseed meal basals for Experiment 5B. Experiment 6. One group of 10 hens 50 weeks of age was assigned to each treatment. Chromium was added as OCI3 • 6 H 2 0 . Data are for the 6 days following a 4 day adjustment period on the experimental diets. Experiment 7. Two groups of 10 hens 35 weeks of age were assigned to each treatment except 4 groups were fed the control diet. The dehydrated grass was added to the basal diet at the expense of ground corn. Where statistical differences in performance are indicated, significance was determined using Duncan's new multiple range test following an analysis of variance that indicated a factor to have significant effects. Where pretest data are given and more than a single pen is assigned a treatment, the analysis was made on the change in value of the factor measured for each
TABLE 6. The effects of ascorbic acid and EDTA1 on vanadium toxicity in laying hens (Experiment 5A) Treatment Vanadium (ppm) 0 40 40 40 40 40
Haugh units
Ascorbic acid
Rate lay
Body weight change
(%)
EDTA
Pretest
4 wk
Change
(%)
(g)
0 0
0 0 0 6X 1 8X 6X
78.6 81.0 80.5 80.7 80.4 81.6
76.5 64.2 78.7 69.1 68.6 81.5
-2.ia -16.8b -1.8a -11.6b -11.8b -.ia
65a 62a 75a 73a 70 a 77a
+17a -15a +4 a -29a _9a +6a
.5 0 0 .5
a' bMeans in a column followed by a common superscript are not significantly different (P>.05). 1 EDTA was added as disodium ethylenediaminetetraacetate at 6 and 8 times the molar concentration of vanadium.
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Specific differences in the various experiments are: Experiment 1, Two groups of 10 hens 50 weeks of age were assigned to each treatment with the experimental diets being fed for 7 weeks. Experiment 2. One group of 10 hens 60 weeks of age was assigned to each treatment. Ration 2A (Table 1) was calculated to be 12%; 2B, 15%; 2C, 20%; and 2D, 25% protein. Experiment 3. Two groups of 10 hens 50 weeks of age were assigned to each treatment. Ration 3 A (Table 1) was the mixed grain basal, 3B the corn-fish basal, and 3C the corn-sucrosefish basal diet. Haugh units were determined on two consecutive eggs per hen per treatment prior to feeding the experimental diets and again after 4 weeks on trial. Experiment 4. One group of 10 hens 48 weeks of age was assigned to each treatment. Haugh units and rate of lay were determined using all eggs laid during the second week of the trial. Experiment 5. Two groups of 10 hens 40
VANADIUM TOXICITY IN LAYING HENS
1157
TABLE 7. The effects of cottonseed meal (CSM) on the vanadium toxicity in laying hens (Experiment 5B) Haugh units
Treatment Vanadium (ppm) Pretest
Basal Soy
40 10% CSM 40 20% CSM 40
78.6 81.0 80.2 81.4 80.4 82.6
4 wk
Change
Rate of lay (%)
Body weight change (g)
76.5 64.2 77.5 71.4 80.4 76.2
-2.1* -16.8b -2.7* -10.0ab 0» -4.4a
65bc 62= 74ab 7jab 77a 77a
+17*b -ISa +60 b +60b +72b +5lb
pen during the test. Where only a single pen is assigned a treatment, the analysis was made using the mean of data for each individual hen for the period indicated. RESULTS AND DISCUSSION Tabulated results for each experiment are given in Tables 2 through 10. In these experiments and in others not reported here, the addition of vanadium at levels of 20 ppm and above quickly (within 3 to 4 days) caused a substantial deterioration in albumen quality. The albumen quality continued to decline for at least 7 weeks. Levels of 80 ppm and above caused near cessation of egg production and considerable molting. Vanadium at 40 ppm usually reduced egg production. Removal of vanadium from the diets allowed albumen quality and egg production to return to normal, the time required depending on the length of the feeding period and the level at which vanadium was fed. Only slight differences in effects were found when the different vana-
dium compounds were compared (Table 8). VOCI2 appeared to be less toxic than VOSO4 orNH4V03. Dietary vanadium at 40 ppm fed for 4 weeks or more also significantly reduced feed intake, body weight, egg weight, and shell quality (Table 2). The effect on egg size and shell quality is contrary to other reports (Berg et al., 1963; Hafez and Kratzer, 1976b). The unique effect is the rapid deterioration in albumen viscosity as measured by Haugh units. The degree of depression of Haugh units and the other effects caused by vanadium depends upon the type of diet and upon certain dietary additives. The use of herring fish meal rather than soybean meal as the supplemental protein source coupled with partial replacement of the grain with sucrose intensified the depression of Haugh units, body weight, and rate of lay (Table 4). This agrees with the effects on growing chicks (Berg, 1966; Hafez and Kratzer, 1976a). Dehydrated grass added at levels from 6 to 12% of the diet prevented the depression of egg production but had little effect on the
TABLE 8. Effect of different vanadium compounds on the performance of laying hens (Experiment 5C) Vanadium Source Control NH„VO., VOCl2
voso4
Haugh units
leve' (ppm)
Pretest
2wk
4 wk
Change
Rate of lay (%)
Body weight change (g)
20 40 20 40 20 40
77.6 77.4 79.6 76.5 78.6 78.4 79.0
78.5 69.6 68.2 71.1 69.7 71.7 68.5
78.7 67.3 65.7 69.9 68.8 72.2 68.3
+l.la -10. l b -13.9b -6.6ab -9.8b -6.2*b -10.7b
74a 68a 57b 69a 71a 70 a 65 a b
+20 a +iab -46b +18 a 0ab +32 a +2ab
Means in a column not followed by a common superscript are significantly different (P<.05).
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' ' Means in a column followed by a common superscript are not significantly different (P>.05).
1158
OUSTERHOUT AND BERG
TABLE 9. Effect of dietary chromium on albumen quality deterioration caused by dietary vanadium in laying hens (Experiment 6) Chromium 0
20 Haugh units
Vanadium
/
81.3 b c go.iabc 76.5 a 77.7ab
^ 83 .OC 79.3abc 76.5a 76.3 a
' ' Means for a parameter not followed by a common superscript are significantly different (P<.05).
maintenance of albumen quality caused by 20 to 40 ppm vanadium (Table 10). In chicks, 5% dehydrated grass almost prevented the growth depression (Berg and Lawrence, 1971). It appears that vanadium acts through different metabolic routes in its effects on albumen quality and on egg production. Dietary protein levels between 12 and 25% obtained by increasing the soybean meal in the diet had no observable effect on the reduction
TABLE 10. Effects of dehydrated grass on vanadium toxicity in laying hens (Experiment 7) Haugh units
Treatment 1
Grass (%) 0
3 6 12 Averages 0 3 6 12
Vanadium (ppm)
Pretest
4 wk
Change
Rate of lay ( 2 - 4 wk) (%)
0 20 40 20 40 20 40 20 40
90.0 88.4 89.5 89.8 90.0 90.2 91.4 91.0 89.8
82.6 74.1 74.2 74.4 68.0 77.6 72.8 78.4 73.4
-7.4a -14.3b -14.3b -15.4b -22.0C -12.6ab -18.6bc -12.6ab -16.4bc
76ab 5 gbcd 40d 76ab 50cd 84a 70abc 7^abc 76ab
20-40 20-40 20-40 20-40 0 20 40
89.0 89.9 90.8 90.4 90.0 89.8 90.2
74.7 71.2 75.2 75.9 82.6 76.1 72.3
-14.3 -17.7 -15.6 -14.5 -7.4a -13.7b -17.9C
49a 63b 77b 73b 76b 70b 59 a
' ' ' Means in a column or group of averages not followed by a common superscript are significandy different (P<.05). 1 Dehydrated grass was added at the expense of ground corn.
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(ppm) 0 20 40 80
in albumen quality caused by 40 ppm vanadium (Table 3). Hill (1979b) had reported a protective effect for higher protein levels with chicks. However, replacing all or at least half of the soybean and meat and bone meal with cottonseed meal almost prevented Haugh unit depression caused by 40 ppm vanadium and increased egg production and body weight over that of the controls (Table 7). Effects of lower levels of cottonseed meal, similar to those that prevented growth depression with chicks (Berg and Lawrence, 1971), had no discernible effects and are not reported here. Because EDTA had been reported by Hathcock et al. (1964) and Hafez and Kratzer (1976a) to alleviate vanadium toxicity in chicks, it was tested singly and in combination with other factors to determine its effects on laying hens. With 20 or 25% protein diets, it protected against a loss of albumen quality. It had no significant effect on egg production or body weight (Tables 3, 4, and 6). Chromium added to the diets at 20 ppm had no detectible effect in preventing the albumen quality deterioration caused by 20 to 80 ppm vanadium (Table 9); this is contrary to the report of Jensen and Maurice (1980). Perhaps the protective effect of chromium takes longer to develop than the 10 days of this trial. Their
VANADIUM TOXICITY IN LAYING HENS trials ran 4 to 6 weeks. Regression coefficients for Haugh units and vanadium levels were: No chromium: Haugh units = 80.6 - .048 x V (ppm), r = - . 7 4 8 20 ppm chromium: Haugh units = 81.6 - .080 X V (ppm), r = - . 8 6 8
the diet and the age and strain of the hens. REFERENCES Berg, L. R., 1966. Effect of diet composition on vanadium toxicity for the chick. Poultry Sci. 45:1346-1352. Berg, L. R., G. E. Bearse, and L. H. Merrill, 1963. Vanadium toxicity in laying hens. Poultry Sci. 42:1407-1411. Berg, L. R., and W. W. Lawrence, 1971. Cottonseed meal, dehydrated grass and ascorbic acid as dietary factors preventing toxicity of vanadium for the chick. Poultry Sci. 50:13991404. Chen, A. A-T., and C. F. Nockels, 1973. The effects of dietary vitamin C, protein, strain and age on egg quality production and serum and albumen protein of chickens. Poultry Sci. 52:1862—1867. Hafez, Y.S.M., and F. H. Kratzer, 1976a. The effect of diet on the toxicity of vanadium. Poultry Sci. 55:918-922. Hafez, Y.S.M., and F. H. Kratzer, 1976b. The effect of pharmacological levels of dietary vanadium on the egg production, shell thickness and egg yolk cholesterol in laying hens and coturnix. Poultry Sci. 55:923-926. Hathcock, J. N., C. H. Hill, and G. Matrone, 1964. Vanadium toxicity and distribution in chicks and rats. J. Nutr. 82:106-110. Hill, C. H., 1979a. Studies on the ameliorating effect of ascorbic acid on mineral toxicities in the chick. J. Nutr. 109:84-90. Hill, C. H., 1979b. The effect of dietary protein levels on mineral toxicity in the chick. J. Nutr. 109: 501-507. Jensen, L. S., and D. V. Maurice, 1980. Dietary chromium and interior egg quality. Poultry Sci. 59:341-346.
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Although the coefficients were not significantly different, there was an indication that the chromium addition improved the Haugh units in the basal diet, but the improvement was quickly lost when vanadium was added. This trial was too short for the expected loss of egg production normally caused by 80 ppm vanadium to be significant. The addition of ascorbic acid at .4 or .5% completely prevented the depression of albumen quality and egg production caused by up to 40 ppm vanadium (Tables 5 and 6). However, dietary ascorbic acid did not prevent the egg weight depression caused by vanadium and appeared to depress the egg weight from the diet without added vanadium. Berg and Lawrence (1971) and Hill (1979a) have reported that .1 to .25% ascorbic acid prevented vanadium toxicity with chicks caused by 20 ppm vanadium. Chen and Nockels (1973) have reported that the effect of ascorbic acid on egg production, egg weight, and Haugh units depends upon the protein content of
1159