Production of chickens with marginal vitamin A deficiency.
ABSTRACT Marginally vitamin A-deficient 1-d-old chickens capable of remaining healthy for at least 6 weeks were produced using a two-generation model. In this model, hens fed on diets with a limited vitamin A content were used to obtain 1-d-old chickens which were marginally deficient in vitamin A. Only hens with a narrow range of plasma retinol values (0.60-0.85 mumol/l) were satisfactory for this purpose. Above this range the 1-d-old chickens were not marginally vitamin A deficient. Below this range egg production and hatchability were affected to some extent depending on the degree of vitamin A deficiency. Even when egg production and hatchability remained at a high level in such birds, the 1-d-old chickens produced were not sufficiently strong to survive the first weeks of life. The advantages of the two-generation model for producing marginally vitamin A-deficient chickens are the increased uniformity and predictability of the chickens with respect to body-weight, general health and vitamin A status. However, it does take about 3 months to produce such chickens.
- SourceAvailable from: nutrition.org[show abstract] [hide abstract]
ABSTRACT: Newcastle disease virus (NDV) infection in chickens differing in vitamin A status has been selected as a model to examine the interrelationship between marginal vitamin A deficiency and the severity of consequences of measles infection in humans. Day-old chickens with limited vitamin A reserves, the progeny of marginally vitamin A-deficient hens, were fed purified diets containing either marginal (120 retinol equivalents/kg diet, ad libitum) or adequate (1200 retinol equivalents/kg diet, ad libitum or pair-fed) levels of vitamin A for a period of 10 wk. At 4 wk of age, half of the chickens in each group were infected intraocularly with the lentogenic, i.e., mildly pathogenic, La Sota strain of NDV. Within 1 wk of infection, plasma retinol levels in the infected, marginally vitamin A-deficient chickens showed a significant and persistent decrease compared to their noninfected counterparts fed the same diet. Moreover, infection with NDV resulted in increased rates of morbidity in the marginally vitamin A-deficient chickens compared with nondeficient chickens. The results of this study indicate that pre-existing marginal vitamin A status increases the severity of disease following NDV infection, and that infection with NDV reduces marginal plasma vitamin A levels to levels which can be regarded as deficient.Journal of Nutrition 07/1989; 119(6):932-9. · 4.20 Impact Factor
- [show abstract] [hide abstract]
ABSTRACT: The effect of Newcastle disease virus (NDV, La Sota strain) infection on vitamin A metabolism was investigated in chickens maintained on normal or marginal vitamin A intake. NDV, a virus of the Paramyxoviridae family that primarily affects epithelial tissue, was administered at 4 wk of age. Plasma levels of retinol, retinol-binding protein and, to a lesser extent, transthyretin were found to be significantly lower during both the acute and postacute phases of infection in chickens fed a diet marginally deficient in vitamin A compared to noninfected birds fed the same diet, while vitamin A content in liver was unaffected. However, in chickens fed adequate vitamin A, NDV infection did not influence the parameters measured. Levels of retinol-binding protein in liver were significantly increased by inadequate vitamin A nutriture, but infection partly reduced this increase. The results suggest that the reduced vitamin A status in marginally vitamin A-deficient chickens infected with NDV can be attributed to a combination of a direct effect of the virus on retinol-binding protein metabolism in liver and an increased rate of utilization and catabolism of retinol and retinol-binding protein by extrahepatic tissues.Journal of Nutrition 07/1989; 119(6):940-7. · 4.20 Impact Factor
- Poultry Science 04/1965; 44:446-52. · 1.52 Impact Factor
British Journal of Nutriiion (1992), 68, 283-291
Production of chickens with marginal vitamin A deficiency
BY CLIVE E. WEST', S. REINDER SIJTSMA'~2~3*,
JAN H. W. M. ROMBOUT* AND AKKE J. VAN DER ZIJPP3$
' Department o f Human Nutrition, Wageningen Agricultural University, PO Box 8129, 6700 EV
Wageningen, * Department of Experimental Animal Morphology and Cell Biology and Department
HARRY P. F. PETERS't,
of Animal Husbandry, Wageningen Agricultural University, PO Box 338, 6700 AH Wageningen,
(Received 19 February 1991 -Accepted 1 August 1991)
Marginally vitamin A-deficient I-d-old chickens capable of remaining healthy for at least 6 weeks were
produced using a two-generation model. In this model, hens fed on diets with a limited vitamin A content
were used to obtain l-d-old chickens which were marginally deficient in vitamin A. Only hens with a
narrow range of plasma retinol values (0-60-0%5 pmol/l) were satisfactory for this purpose. Above this
range the I-d-old chickens were not marginally vitamin A deficient. Below this range egg production and
hatchability were affected to some extent depending on the degree of vitamin A deficiency. Even when
egg production and hatchability remained at a high level in such birds, the l-d-old chickens produced
were not sufficiently strong to survive the first weeks of life. The advantages of the two-generation model
for producing marginally vitamin A-deficient chickens are the increased uniformity and predictability of
the chickens with respect to body-weight, general health and vitamin A status. However, it does take
about 3 months to produce such chickens.
Vitamin A deficiency: Chicken: Animal model
Animal models have often been used in vitamin A research (Scrimshaw et al. 1968; Sporn
et al. 1984). However, many problems can arise in producing vitamin A deficiency in
animals (Underwood, 1984; Beisel, 1988). Sometimes animals have to be fed diets deficient
in vitamin A for a long time before the deficient state is reached and often large variations
in vitamin A status can be observed within the same diet group. In addition, diets free of
vitamin A can produce sudden and uncontrollable vitamin A deficiency with consequent
loss of appetite and concomitant protein%nergy malnutrition. Signs of vitamin A
deficiency are sometimes irreversible, and together with increased sensitivity to infection,
can lead to death. A few models have been described in which many of these problems have
been overcome (Stowe et al. 1980; Davis & Sell, 1983; Nauss et al. 1985; Pueng-
tomwatanakul & Sirisinha, 1986; Smith et al. 1987). One system which can be used is a
two-generation model in which parent animals are made marginally deficient in vitamin A
(Stowe et al. 1980; Nauss et al. 1985; Smith et al. 1987). Their progeny are also marginally
vitamin A deficient and can be used in studies requiring young animals.
The aim of the present study was twofold : first, to produce l-d-old chickens marginally
deficient in vitamin A for a long period without health problems; second, to document the
advantages of such a two-generation model over a one-generation model based on 1-d-old
chickens with an adequate vitamin A status.
Present addresses * Hendrix Voeders, PO Box I, 5830 MA Boxmeer; t Department of Medical Physiology and
Sports Medicine, University of Utrecht, Vondellaan 24, 3521 GG Utrecht; $Research Institute for Animal
Production 'Schoonoord', PO Box 501, 3700 AM Zeist, The Netherlands.
N U T 68
C. E. WEST A N D OTHERS
MATERIALS A N D METHODS
Animals, diets and experimental design
White Leghorn laying hens (Lohmann Selected Leghorn strain), aged approximately 18
weeks, were obtained from a commercial breeder (Verbeek, Barneveld). On arrival, the
hens were housed individually in a room controlled for temperature (20"), relative humidity
(4&50 %) and light-dark cycles (I6 h dimmed light - 8 h darkness). A commercial diet
(Opfokvoer 2, Rijnzate, Wageningen) was fed for 1 week. After this period the birds were
allocated randomly to five dietary groups, each of twelve hens. The first four groups (P-0,
P-300, P-600 and P-1500) were fed on purified diets varying in vitamin A content (0, 300,
600 or 1500 retinol equivalents (RE)/kg respectively) and the fifth group (N- 1500) was fed
on a diet based on non-purified components with the same amount of vitamin A as in the
fourth group (Table 1). This fifth group was added in order to test for possible differences
between purified and non-purified diets. Water and diets were provided ad lib. and the hens
had free access to oyster shell grit. The laying hens were studied for a total period of 19
weeks during which time feed consumption, body-weight, egg production and plasma
retinol concentration were measured. Hatchability and health status of I-d-old chickens
were determined three times during the experiment (weeks 8, 13 and 18).
In this experiment, two groups were formed from progeny of marginally vitamin A-
deficient hens. These hens were fed on diets containing either 300 or 600 RE/kg for at least
13 weeks, resulting in plasma retinol levels of 0.35-0.50 pmol/l and 0.60-0.85 pmol/l
respectively. Both groups of chickens were fed on a purified diet limited in vitamin A
(120 RE/kg; Table I). In addition, female I-d-old chickens (Lohmann Selected Leghorn
strain) were obtained from a commercial hatchery (Verbeek, Barneveld). These chickens
were allocated to a further three groups. One of those groups was also fed on a purified diet
limited in vitamin A (120 RE/kg); another was fed on a purified diet free of vitamin A; a
third group was also fed on the purified diet free of vitamin A for 2 weeks and then fed on
the diet limited in vitamin A (120 RE/kg). The 1-d-old chickens, twelve birds/group and
one group/cage, were housed as described previously (Sijtsma et a/. 1989~). They were
studied for a period of 6 weeks during which health status and plasma retinol levels were
Sampling of blood
Blood from a wing was collected in heparinized tubes; after centrifugation, plasma was
stored at -20".
Vitamin A in plusma
In Expt 1, retinol levels were determined in the plasma of hens at weeks 0, 5, 8, 1 I, 13, 15,
17 and 19, and in plasma from their I-d-old progeny. In Expt 2, retinol analyses in the
plasma of chickens were carried out at 0, 2, 3.54, and 5-6 weeks. A reversed-phase high-
performance liquid chromatographic (HPLC) method modified from that of Driskell et al.
(1982) was used, as described previously (Sijtsma et al. 1989a, b).
Vitamin A content of diets
In order to determine the content and stability of vitamin A in diets used in Expt 1, analyses
were carried out at 1 and 10 weeks after manufacture as described by Manz & Philipp
MARGINAL VITAMIN A DEFICIENCY IN CHICKENS
Table 1. Composition (g/kg) of the purified and non-purijied diets*
Diet . . .
Vitamin-mineral premix§, 7
- - _ _ _ ~ -
* Diets were manufactured in pelleted form following the guidelines of the National Research Council (1984)
by the Institute of Animal Nutrition and Physiology (IGMB-TNO, Wageningen). The metabolizable energy of the
purified hen and chicken diets, and the non-purified hen diet was 12.54 (3000), 13.38 (3200) and 12.44 (2975) MJ
(kcal)/kg feed, respectively.
t Soya-bean isolate: Purina Protein 500 E, Ralston Purina, St Louis, MO, USA, containing 880 g protein/kg
$ The laying hens had free access to oyster shell grit.
9 The vitamin and mineral premix (10 g/kg purified diet) was prepared with dextrose and contained (mg/kg
diet): thiamin 2.5, riboflavin 5.5, pantothenic acid 15.0, nicotinic acid 50.0, pyridoxine 5.0, biotin in hens 0.10 and
in chickens 015, folic acid in hens 0.45, and in chickens 075, choline chloride in hens 1000 and in chickens 1850,
cyanocobalamin 0.01 5, inositol 100, p-aminobenzoic acid 50.0, cholecalciferol 0.075, m-a-tocopherol 30.0,
menadione 50, L-ascorbic acid 50.0, FeSO,. 7H,O 400, MnO, 150, CuSO,. 5H,O 100, ZnSO,. H,O 200,
Na,SeO,.SH,O 0.3, KI 5, ethoxyquin 100. Vitamin A was added as retinyl acetate and retinyl palmitate, Rovimix
A 500 (156.6 mg retinol/g; F. Hoffmann-La Roche, Basel, Switzerland) in hens: 0, 300, 600 or 1500 retinol
equivalents/kg diet, and in chickens: 0 or 120 retinol equivalents/kg diet.
11 The vitamin and mineral premix (10 g/kg non-purified diet) was prepared with soya-bean flour and contained
(mg/kg diet): riboflavin 5.5, pantothenic acid 15.0, nicotinic acid 30.0, pyridoxine 2.0, biotin 0.05, folic acid 0.1,
choline chloride 500, cyanocobalamin 0.01 5, cholecalciferol 0,075, DL-a-tocopherol 30.0, menadione 5.0, L-
ascorbic acid 50.0, FeSO, .7H,O 400, MnO, 150, CuSO, .5H,O 100, ZnSO,. H,20 200, Na,SeO,. 5H,O 0.3, KI 2,
ethoxyquin 100. Vitamin A was added as retinyl acetate and retinyl palmitate, Rovimix A 500, 1500 retinol
7 Calculation of the retinol and carotene content was based on the amount of vitamin A added to the diet and
on feedstuff analysis tables respectively (Agricultural Research Council, 1975; Allen, 1984; National Research
Council, 1984). All the carotene present was assumed to be p-carotene and the values were divided by 6 to convert
to retinol equivalents. In the N-1500 diet, the p-carotene content was calculated to be 60 /Lg RE/kg, while in the
other diets no p-carotene was present. Analysis of the hens’ diets 1 week after preparation showed that vitamin
A content was more than expected: 420, 720, 1830 and 2040,ug RE/kg for P-300, P-600, P-1500, and N-1500
respectively. After 10 weeks of storage the content decreased to 240, 570, 1790 and 1620 pg RE/kg for P-300, P-
600, P-1500 and N-I500 respectively. No vitamin A was detected in the P-O diet, 1 or 10 weeks after manufacture.
Tapioca (650 g/kg)
Soya-bean flour (50 g/kg)
Vitamin-mineral premix jl,l
All statistical comparisons among treatment groups were performed by one-way analysis
of variance after testing for normality. Differences between group means were evaluated by
Tukey’s range test (Winer, 1971).
C. E. WEST A N D OTHERS
Table 2. Expt 1. Hatchability o f eggs laid by hens fed on diets varying in viramin A
Proportion of eggs laid
Dietary group.. .
P-300 P-600 P- 1 500
P- 1 500
Embryos surviving, 8 d
Embryos surviving, 17 d
Embryos surviving, 8 d
Embryos surviving, 17 d
Embryos surviving, 8 d
Embryos surviving, 17 d
~ _ _ _ _ _
100 (1 10)
P-0, P-300, P-600, P-1500, purified diets containing 0, 300, 600 and 1500 retinol equivalents (RE)/kg
respectively; N-l 500, non-purified diet containing I 500 RE/kg.
* Eggs collected over 11 d were hatched in three successive periods starting in weeks 8, 13 and 18. For
t The numbers of eggs incubated are shown in parentheses. Numbers exclude eggs that were cracked or
contained double yolks.
$ Hens were removed before the third egg collection period.
description of diets, see Table 1.
GeneraI health o f laying hens
Health status clearly deteriorated in the P-0 and P-300 groups. Morphological signs of
vitamin A deficiency were seen in three hens that died in these groups. After 14 weeks it was
decided, for ethical reasons, to remove birds from the P-0 group. Significant differences in
body-weight among the groups could not be observed throughout the experiment (values
not shown). From weeks 11-12, feed consumption and egg production of the P-0 group
were significantly lower than those of the P-1500 group. Hens fed on a purified diet
adequate in vitamin A and their counterparts fed on a non-purified diet with the same
vitamin A level were similar with respect to body-weight, feed consumption and egg
production (values not shown).
Hatchability was not affected in the first period (Table 2) and the I-d-old chickens
produced were in a healthy condition. However, in the second period, hatchability was
poor in the P-0 group. Many embryos died during the first 8 d of incubation, while others
were too weak to break out of the shell. In the third period, hatchability tended to be lower
in the P-300 group than in the other groups. Differences in weight of eggs or body-weight
of 1-d-old chickens could not be observed among the diet groups in all periods measured,
while weight of eggs and body-weight of 1-d-old chickens increased slightly from the first
to the third period (values not shown).
MARGINAL VITAMIN A DEFICIENCY IN CHICKENS
0 2 4
Duration of dietary treatment (weeks)
10 12 14 16 18
Fig. I. Expt I. Plasma retiiiol concentration of laying hens fed on diets varying in vitamin A content. For
description of diets, see Table 1. Points represent ineans with their standard errors varying from 1 to 10% for ten
to twelve hens. (O--. O), P-0; (+), P-300; (0-O), P-600; ( . - .
Marginal vitamin A status (0.35-0.70 /tmol/l); above this area, vitamin A status is normal and below this area it
is deficient (< 0.35 pol/l). P-0, P-300. P-600, P-1500, purified diets containing 0, 300, 600 and 1500 retinol
equivalents (RE)/kg respectively; N- 1500, non-purified diet containing 1500 RE/kg.
.-a). P-1500; (*-*),
Table 3. Evpt 1. Plasma retinol concentration in I-d-old chickens derived as progeny from
hens fed on diets varying in vitamin A content
(Mean values with their standard errors for the progeny of six hens/group Plasma samples from all
1-d-old chickens from one hen were pooled)
_ _ _ _ _ _ _ _ _ - _ _ _ - _ _ _ _
Retinol concentration @mol/l)
- _ _ _ -
_ - - - - - - - -
_ - - - - ___ -
P-0, P-300. P-600, P-1500, purified diets containing 0, 300, 600 and I500 retinol equivalents (RE)/kg
respectively, N- 1500. non-purified diet containing 1500 RE/kg , ND, not determined
Means within a column with unlike superscript letters were significdntly different (P < 005, Tukey)
* For description of diets, see Tdble 1
_ _ _ _ _ _ _ _ _ _ - _ _ _ _ _ _ _ _ _ _
_ _ _ - _
Plasma retinol concentration
At the start of the experiment mean plasma retinol levels of the 18-week-old hens ranged
from 2.4 to 3.0 pmol/l (Fig. 1). These levels decreased in all groups, including the P- 1500
and N-1500 groups throughout the experiment. The fall in plasma retinol levels was
negatively related to the level of vitamin A in the diet. Plasma retinol concentrations in 1-
d-old chickens derived from these hens are shown in Table 3. In the first hatching period
plasma retinol levels were similar in all groups (between 0.86 and 1.0.5,~~mol/l).
progeny of the P-300 group the concentrations were significantly lower than in the progeny
of all other groups after the second period (0.1 1 pmol/l compared with 0.89, 1.05 and
1.14 ,umol/l). Similar results were obtained after the third period for the progeny of the P-
600 group. Plasma retinol concentrations in the progeny of the P- I500 and N- I500 groups
were stable throughout the experiment.