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Effects of Methionine and Valine on Growth and Antibody Production in Chicks Infected with Live or Killed Newcastle Disease Virus
614
C. E. FSANTI, L. M. JULIAN, H. E. ADLER AND L. Z. MCFARLAND Poultry Sci. 44: 835-838. Reyniers, J. A., M. Wagner, T. D. Luckey and H. A. Gordon, 1960 Survey of germ-free animals: the White Wyandotte bantam and White Leghorn chicken. Lobound Reports No. 3, Univ. Press, Notre Dame, Ind. 7-171. Tindell, L. D., C. H. Moore, N. R. Gyles, W. A. Johnson, L. J. Dreesen, G. A. Martin and P. B. Siegel, 1967a. Genotype-environment interactions in broiler stocks of chickens. 1. The importance of stock by location and stock by trial interactions. Poultry Sci. 46: 603-611. Tindell, L. D., C. H. Moore, N. R. Gyles, H. R. Wilson, W. A. Johnson, L. J. Dreesen, W. L. Blow, W. F. Krueger and P. B. Siegel, 1967b. Genotype-environment interactions in egg production stocks of chickens. 1. The importance of stock by location and stock by year interactions. Poultry Sci. 46: 1077-1088.
Effects of Methionine and Valine on Growth and Antibody Production in Chicks Infected with Live or Killed Newcastle Disease Virus K. K. BHARGAVA, R. P. HANSON AND M. L. SUNDE Departments oj Poultry Science and Veterinary Science, University of Wisconsin, Madison, Wisconsin 53706 (Received for publication September 21. 1970)
I
N PREVIOUS experiments (Bhargava et al., 1970) it was reported that the antibody titer was higher for chicks receiving 0.3 to 0.6% L-methionine levels than for chicks receiving 0.7 to 1.1% L-methionine. The optimum requirement of methionine for growth was about 0.7% while for antibody production it was considerably lower. The reverse was true with valine. As the level of valine in the diet was increased, the antibody titer increased. The requirement of valine for maximum growth response was 0.9% but for optimum antibody production it was higher. Gill and Published with the approval of the Director of the Wisconsin Agricultural Experiment Station, College of Agricultural and Life Sciences, Madison.
Gershoff (1967), using synthetic antigen, have reported that antibody production was increased by methionine deficiency and decreased by excess methionine in monkeys. However, in 1968, Gershoff et al. did not observe any change in antibody production in rats due to deficiency or excess of methionine using synthetic antigen or sheep red blood cell antigen. Ishiguro and Koyanagi (1968) observed that the rats fed a diet containing 0.28% methionine showed 8.3% gamma globulin, while it was 7.3% for rats fed 0.58% of methionine in the diet. It is possible that the different levels of specific amino acid in the diet may either directly affect the production of antibodies, or may affect the replication of virus which
Downloaded from http://ps.oxfordjournals.org/ at Osaka Daigaku Ningen on June 24, 2015
tionships to body measurements of commercial strains of turkeys. Poultry Sci. 48: 1598-1603. Oakberg, E. F., 1951a. Genetic differences in quantitative histology of the adrenal, organ weights, and inter-organ correlations in White Leghorn chickens. Growth, 15: 57-78. Oakberg, E. F., 1951b. Influence of genetic constitution on growth of lymphoid tissue in liver and pancreas of White Leghorn chickens, and correlation of lymphoid tissues with weights of some visceral and endocrine organs. Growth, 15: 79-100. Oakberg, E. F., and A. M. Lucas, 1949. Variations in body weight and organ: body weight ratios of inbred lines of White Leghorn chickens in relation to mortality, especially from lymphomatosis. Growth, 13: 319-337. Roberson, R. H., and D. W. Francis, 1965. Anatomical development of White Chinese geese.
615
METHIONINE, VALINE AND NEWCASTLE
in turn may influence the gamma globulin production. In order to separate the two processes, the effect of methionine and valine on antibody production in chicks was studied using formalin killed Bl strain of Newcastle disease virus (NDV) as antigen. PROCEDURE
TABLE 1.—Basal diet Amino acid mix1 2
Salts X Vitamin mix3 Anti-acid4 Choline chloride Alpha tocopheryl acetate, 10 mg./gm. Cellulose NaHC0 3 Corn oil Dextrin to 1 kg.
Variable 60.0 5.0 10.0 2.0 1.0 30.0 10.0 90.0
Downloaded from http://ps.oxfordjournals.org/ at Osaka Daigaku Ningen on June 24, 2015
Day-old chicks, the progeny of New + Hampshire males and Single Comb White 1 Leghorn females were used in all the experAmino acid mix supplied the following in gm./ iments. All chicks were wing banded, kg. of diet: L-arginine, 20; L-glutamic acid, 70; glycine, 20; L-histidine, 6; L-isoleucine, 10; L-leucine, weighed individually and maintained in 20; L-lysine, 16; L-phenylalanine, 10; L-tryptophan, electrically heated batteries with raised 3; L-tyrosine, 9; L-valine, 11 or L-methionine, 8; L-threonine, 9. 2 wire floors. Ten chicks were used per treatSalts X supplied the following in mg./kg. of diet: CaCOs, 15,000; Ca 3 (P0 4 ) 2 , 13,950; KH 2 PO„, ment. The experimental diets and water 12,000; Na 2 HP0 4 , 7,200; NaCl, 6,000; MgS0 4 were given ad libitum from the first day af- •7H 2 0, 5,000; KC1, 3,000; MnS0 4 -2H 2 0, 420; FeC 6 H 5 0 7 , 400; ZnCOs, 200; Kl, 52; CuS0 4 , 20; ter hatching. The individual body weights NaBr, 10; NaMo0 4 , 10. 3 and group feed consumption were recorded Vitamin mix supplied in mg./kg. of diet: thi100; niacin, 100; riboflavin, 16; calcium panat the end of each week. Each experiment amine, tothenate, 20; vitamin Bi2, .02; pyridoxine HC1, 6; was terminated at the end of 18 days. biotin, .6; folic acid, 4; inositol, 100; menadione, 5; A (250,000 I.U./gm.), 40; vitamin D 3 , When the chicks were four days of age, vitamin (200,000 I.U./gm.), 5. 4 Anti-acid 3 parts magnesium trisilicate and 1 they were infected by instilling one drop of 1CT3 solution of a stock solution of Bl part aluminum hydroxide. strain of NDV in each nostril or by injecting O.S ml. per bird of 24-hour formalin in- maintained on practical diet throughout activated NDV per day into breast muscles each experiment to serve as a negative confor five days. Two weeks later (day 18), trol. the blood was obtained from the chicks by The data on weight of chicks and HI tiheart puncture and serum was collected in ter were statistically analyzed by the use of separate sterile vials. The Beta procedure Duncan's multiple range test (Steel and of hemagglutination inhibition test (Poul- Torrie, 1960). try Biologies, National Research Council, RESULTS AND DISCUSSION 1963) was used to determine the antibody titer. In a preliminary experiment, live Bl The composition of basal diet is shown strain of NDV was exposed to both 0.2 and in Table 1. The diets for methionine or va- 0.8% formalin solutions for inactivation eiline experiments were prepared by using ei- ther for 24 hours or 8 days. The inactivather methionine or valine at the prescribed tion of the virus was tested by injecting level and adding the other amino acid in (0.1 ml. per egg) into the allantoic cavity of the amounts indicated in the table. nine-day embryonated eggs and observing In addition to the groups on amino acid mortality of embryos. The results showed diets, one group of ten chicks was fed on a that 24 hours of inactivation with 0.2% practical chick starter diet (Bhargava et formalin was sufficient to inactivate the vial., 1970) and injected with killed virus. A rus. non-infected group of ten birds was also The object of the next experiment was to
616
K. K. BHARGAVA, R. P. HANSON AND M. L. SUNDE
TABLE 2.—Effect of dose and route of inoculation with killed NDV on antibody titers Experiment 2 Group
Inactivated Virus
Route
Mean H I Titer 1 (18 days)
Undiluted 10"' Undiluted 10"1
I/M I/M I/P I/P
1.20 0.50 1.20 0.33
TABLE 3.—Effect of dose and time of inoculation with killed NDV on antibody liters Experiment 3 Group #
Inactivated Virus
Day of Bleeding
1 2 3 4
Undiluted Undiluted IO-1 10"1
18 23 18 23
, H T1 Ti il ttPe ,ri M M ee a nn H 5. SO"2 5.66* 3.50b 3.16 b
1 Values are expressed as log2 of the reciprocal of last dilution. 2 Numbers with different superscript letters are significantly different (P < .05) from other numbers in the same column.
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determine the dose and route for inoculating chicks [fed practical chick starter diet (Bhargava et al, 1970)] with the inactivated virus. Two groups of six chicks each were injected intramuscularly (0.5 ml. per bird) both with undiluted and 10_1 dilution of a stock solution of 24 hours inactivated NDV for five days. The other two groups of six chicks each received intraperitoneal injections of the same dose of killed virus. Table 2 shows that the antibody response obtained with undiluted virus given intramuscularly was higher than the diluted virus (10_1) injected intraperitoneally. Since the antibody titers were very low, it was decided to hyperimmunize the chicks in subsequent experiments by injecting killed virus for five consecutive days to reach titers similar to those obtained in previous experiments. A third experiment was designed to determine the day on which the birds fed practical diet (Bhargava et al., 1970) should be bled (18th or 23rd day) in the subsequent experiments because they received the antigen for five consecutive days. The results of antibody titers in Table 3 show that varying the day of bleeding from the 18th to 23rd day did not make any significant difference in the antibody titers. The antibody titers of birds injected with undiluted killed virus were significantly (P < .05) higher than those receiving lO^1 dilution of the same virus. Since there was no significant difference between the HI titers on the 18th and 23rd day, ti-
ters in subsequent experiments were measured only on the 18th day with an appreciable saving in the diet cost. Two experiments (4 and 5) were then designed with amino acid diets containing the levels of methionine or valine as shown in Tables 4 and 5, a low level, the requirement level and one greater than the requirement were selected for each amino acid. The results (Tables 4 and 5) show that with an increase in the level of methionine or valine, better growth and feed efficiency were obtained. The requirement of methionine for maximum growth response in chicks on this amino acid diet appears to be about 0.7%, which is in agreement with the results obtained by Marrett et al. (1964), Dobson et al. (1964) and Bhargava et al. (1970). As valine was increased in the diet there was an improvement in the average weight and feed efficiency. The optimum requirement for growth with valine seems to be at least 0.9% which agrees with our previous results (Bhargava et al., 1970) and is close to the recommendations of the National Research Council (1966). The results also show that L-methionine induced a greater response than D-methionine at the 0.7% level with respect to chick growth and feed utilization. Tables 4 and 5 summarize the results on
METHIONINE, VALINE AND NEWCASTLE TABLE 4.—Effects
of methionine and valine on growth, feed efficiency and antibody production in chicks Experiment 4 Killed Virus
Diet
0.4% 0.7% 0.7% 1.1%
L-meth. L-meth. D-meth. L-meth.
. , N T-/.-M ; 0 W i" ^ a ,r. . 18 days 0-18 days
Live Virus
Geometric mean HI titer" l g dayg
65. I"3 122.21" 99.9" 143.0 d
5.10 2.00 2.35 1.90
3.88" + 0.35 4 3.88" + 0.31 3.60" + 0.44 3.70" + 0.39
71. > 138.7 d 142.8 d 103.0"
4.65 1.86 1.85 2.15
3.20"+ 3.20» + 4.00" + 3.80" +
0.32 0.32 0.36 0.32
.
AV
. , N ,-, ; r . ; 0 W J- «-» „ U*. 18 days 0-18 days
Geometric mean H I titer 2 l g dayg
4.82
4.40» + 0.37 4
14'.1*
1.95
3.30 b ±0.37
71.7"
4.50
3.20"±0.57
151.8*
1.80
4.20" + 0.32
77.4*3
1
Feed consumed/gain in weight. Values are expressed as log2 of the reciprocal of last dilution. Numbers with different superscript letters are significantly different (P<.05) from other numbers obtained in the respective experiment. 4 Mean + SEM. 2 3
antibody titers in chicks infected or injected with live or killed virus. The antibody titers in experiment 4 were lower as compared to the titers observed in previous experiments, although the differences were nonsignificant (P > .05), except in chicks fed methionine diets and infected with live virus. Table 5 shows that the HI titer was found to be significantly higher (P < .05) for 0.4% L-methionine group than for groups receiving 0.7 to 1.1% L-methionine, TABLE 5.—Effects
irrespective of the killed or live virus which they received. The antibody titer was slightly higher for L-methionine than D-methionine, but the difference was not found to be significant (P < .05). The HI titer was low in chicks fed a low level (0.5%) of valine and it increased significantly (P < .05) at higher levels (1.5%) of valine in the diet. The antibody titers of chicks (infected with live virus) fed graded levels of methionine in the diet showed a significant
of methionine and valine on growth, feed efficiency and antibody production in chicks Experiment 5 Killed Virus
Diet
0.4%, 0.7% 0.7% 1.1%
L-meth. L-meth. D-meth. L-meth.
0.5% Val. 0.9% Val. 1.5% Val. Pract. diet 1
Av.wt.(g.) F/& 18 days 0-18 days 65.2» 135.4" 90.3 b 135.5" 83.6b 137.7" 138.2" 171.4 d
3
Ge0
Live Virus g T e t t r LT a n ™ titer
5.31 2.08 2.96 1.90
5.22" + 0.22" 4.22»b + 0.40 4.2O b + 0.41 4.00 b + 0.36
3.81 2.00 1.90 2.00
2.80"±0.40 3.30" b + 0.49 4.80 b + 0.35 4.00 b ±0.36
Av.wt.(g.) l g days
F/G „_„ ^
^
S ^ 1 H I titer
67.8"
5.28
5.44"±0.42
139.8"
1.96
4.00b±0.25
80.2 b
4.27
3.00" + 0.25
d
1.80
5.20 b ±0.38
164.2
1
Feed consumed/gain in weight. Values are expressed as log2 of the reciprocal of last dilution. 3 Numbers with different superscript letters are significantly different (P<.05) from other numbers obtained in the respective experiment. 4 Mean + SEM. 2
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0.5% Val. 0.9% Val. 1.5% Val. Pract. diet
.
AV
617
618
K. K. BHAEGAVA, R. P. HANSON AND M. L. SUNDE
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(P < .05) difference in both experiments, onine decreases. and with valine the differences were signifiThe low or high antibody titers in chicks cant in experiment 5 and approached sig- fed low or high levels of valine in the diet nificance in experiment 4. These results are parallel to the observations of Koyanagree rather well with the results obtained agi and Ishiguro (1966) and Gershoff et al. in previous experiments (Bhargava et al., (1968) on amounts of gamma globulin syn1970). The chicks immunized with forma- thesized in rats fed diets containing low or lin killed virus and fed graded levels of me- high levels of tryptophan and phenylalathionine and valine showed antibody re- nine. Our results can also be explained on sponse which was similar, but slightly the basis of the fact that chicken gamma lower than the titers obtained in chicks in- globulin has a high valine content (Tenenfected with live virus. These results show house and Deutsch, 1966). that the different levels of methionine or SUMMARY valine in the diet appear to have a major influence upon antibody production and Day-old chicks were fed practical or possibly a minor one on the virus replica- amino acid diets. The effects of methionine tion. or valine deficiency or excess on antibody The results on antibody titers in chicks formation were studied in an otherwise fed methionine diets reported here using complete diet. At the end of day four, the killed or live Newcastle disease virus are chicks were injected or infected with forin agreement with the observations made malin killed or live Bl strain of Newcastle by Gill and Gershoff (1967) in monkeys, disease virus. A single dose (0.5 ml. per Gershoff et al. (1968) in rats. Ishiguro and bird) of antigen produced very low antiKoyanagi (1968) have also obtained simi- body titers, but with five injections of the lar data in studies on the effects of dietary same antigen, antibody titers increased methionine on production of gamma globu- considerably. The undiluted preparation of killed virus produced a highly significant lin in rats. The high antibody titers of chicks fed improvement in antibody titers as com_1 low levels of methionine in the diet may be pared to 10 dilution of the same virus. related to the fact that a low methionine The antibody titer was higher in chicks fed intake might have decreased the albumin 0.4% L-methionine than in chicks receiving content, but not the gamma globulin con- 0.7 and 1.1% L-methionine, irrespective of tent of serum as suggested by Ishiguro and whether the virus was live or killed. The Koyanagi (1968). In addition, the dietary growth rate, and feed efficiency improved need for methionine in antibody production with an increase in the level of methionine appears to be very low. This is in agree- in the diet. L-methionine induced a better ment with the observations of Tenenhouse response with respect to growth, feed utiliand Deutsch (1966) in which they re- zation and antibody production than D-meported that methionine is a very minor thionine. At a low level (0.5%) of valine in amino acid component of chicken gamma the diet, the antibody titer was low, but at globulin. However, the change in albumin- a high level (1.5%) it was high in chicks gamma globulin ratio and the fact that me- receiving either killed or live virus. An imthionine is a very minor constituent of provement in growth, feed efficiency, and gamma globulin do not adequately explain antibody production was observed with an the observation that antibody titers for increase in the level of dietary valine. The NDV increase as the dietary level of methi- different levels of methionine or valine in
METHIONINE, VALINE AND NEWCASTLE
the diet appear to have a major influence upon antibody production and possibly a minor influence on the virus replication. The dietary need of methionine and valine for antibody production appears to be very low and high respectively in comparison to the requirements for optimum growth. REFERENCES
Ishiguro, K., and T. Koyanagi, 1968. Effects of dietary tryptophan and methionine on protein in the serum and liver of rats. Tohoku. J. Exp. Med. 96: 345-348. Koyanagi, T., and K. Ishiguro, 1966. Dietary tryptophan and blood gamma globulin. Tohoku. J. Exp. Med. 90: 245-248. Marrett, L., H. R. Bird and M. L. Sunde, 1964. The effects of different isomers of methionine on growth of chicks fed amino acid diets. Poultry Sci. 4 3 : 1113-1118. National Research Council, 1963. Methods for the examination of poultry biologies. National Academy of Sciences—National Research Council, Washington, D.C., Publ. 70S. National Research Council, 1966. Nutrient requirements of poultry. National Academy of Sciences —National Research Council, Washington, D.C., Publ. 1345. Steel, R. G. D., and J. H. Torrie, 1960. Principles and Procedures of Statistics. McGraw-Hill Book Company, Inc., New York, p. 112. Tenenhouse, H. S., and H. F. Deutsch, 1966. Some physical-chemical properties of chicken gamma globulins and their pepsin and papain digestion products. Immunochemistry, 3 : 11-20.
Effects of Bird Density, Nutrient Density and Perches on the Performance of Caged White Leghorn Layers* R . W . DORMINEY AND G. H . ARSCOTT Department oj Poultry Science, Oregon State University, Corvallis, Oregon 97331 (Received for publication September 21, 1970)
studies have compared cage S EVERAL size and bird density with 1 to S hens per cage (Lowe and Heywang, 1964; Wilson et al, 1964, 1967; Blount, 1965; Moore et al, 1965; Owings et al, 1967; Ruggles et al, 1967; Champion and Zindel, 1968; Shirley, 1969; Ostrander and Young, 1969). Bramhall et al. (1966) and Quisenberry (1968) summarized several studies and concluded that small cages with 2 to S hens per cage providing 350 to 465 square centimeters (sq. cm.) per hen are * Oregon Agricultural Experiment Technical Paper No. 2946.
Station,
usually more suitable than larger cages providing similar space per bird. Most studies show that 1 hen per cage usually performs best, but beyond that the results vary. There is often a decline, either significant or non-significant, in egg production and an increase in mortality as the space per bird decreases. Wilson et al. (1964, 1967) found only part of the increase in mortality to be due to increased cannibalism. Body weight often decreases as the bird density increases, while egg size is usually not affected. Feed consumption per henday was no different with 1, 2, 3 or 4 heavy type layers in 30.5 X 45.7 cm. cages (Rug-
Downloaded from http://ps.oxfordjournals.org/ at Osaka Daigaku Ningen on June 24, 2015
Bhargava, K. K., R. P. Hanson and M. L. Sunde, 1970. Effects of methionine and valine on antibody production in chicks infected with Newcastle disease virus. J. Nutr. 100: 241-248. Dobson, D. C , J. O. Anderson and R. T. Warnick, 1964. A determination of the essential amino acid proportions needed to allow rapid growth in chicks. J. Nutr. 82: 67-75. Gershoff, S. N., T. J. Gill, III, S. J. Simonian and A. I. Steinberg, 1968. Some effects of amino acid deficiencies on antibody formation in the rat. J. Nutr. 9S: 184-189. Gill, T. J., I l l , and S. N. Gershoff, 1967. The effects of methionine and ethionine on antibody formation in primates. J. Immunol. 99 : 883-893.
619
C. E. FSANTI, L. M. JULIAN, H. E. ADLER AND L. Z. MCFARLAND Poultry Sci. 44: 835-838. Reyniers, J. A., M. Wagner, T. D. Luckey and H. A. Gordon, 1960 Survey of germ-free animals: the White Wyandotte bantam and White Leghorn chicken. Lobound Reports No. 3, Univ. Press, Notre Dame, Ind. 7-171. Tindell, L. D., C. H. Moore, N. R. Gyles, W. A. Johnson, L. J. Dreesen, G. A. Martin and P. B. Siegel, 1967a. Genotype-environment interactions in broiler stocks of chickens. 1. The importance of stock by location and stock by trial interactions. Poultry Sci. 46: 603-611. Tindell, L. D., C. H. Moore, N. R. Gyles, H. R. Wilson, W. A. Johnson, L. J. Dreesen, W. L. Blow, W. F. Krueger and P. B. Siegel, 1967b. Genotype-environment interactions in egg production stocks of chickens. 1. The importance of stock by location and stock by year interactions. Poultry Sci. 46: 1077-1088.
Effects of Methionine and Valine on Growth and Antibody Production in Chicks Infected with Live or Killed Newcastle Disease Virus K. K. BHARGAVA, R. P. HANSON AND M. L. SUNDE Departments oj Poultry Science and Veterinary Science, University of Wisconsin, Madison, Wisconsin 53706 (Received for publication September 21. 1970)
I
N PREVIOUS experiments (Bhargava et al., 1970) it was reported that the antibody titer was higher for chicks receiving 0.3 to 0.6% L-methionine levels than for chicks receiving 0.7 to 1.1% L-methionine. The optimum requirement of methionine for growth was about 0.7% while for antibody production it was considerably lower. The reverse was true with valine. As the level of valine in the diet was increased, the antibody titer increased. The requirement of valine for maximum growth response was 0.9% but for optimum antibody production it was higher. Gill and Published with the approval of the Director of the Wisconsin Agricultural Experiment Station, College of Agricultural and Life Sciences, Madison.
Gershoff (1967), using synthetic antigen, have reported that antibody production was increased by methionine deficiency and decreased by excess methionine in monkeys. However, in 1968, Gershoff et al. did not observe any change in antibody production in rats due to deficiency or excess of methionine using synthetic antigen or sheep red blood cell antigen. Ishiguro and Koyanagi (1968) observed that the rats fed a diet containing 0.28% methionine showed 8.3% gamma globulin, while it was 7.3% for rats fed 0.58% of methionine in the diet. It is possible that the different levels of specific amino acid in the diet may either directly affect the production of antibodies, or may affect the replication of virus which
Downloaded from http://ps.oxfordjournals.org/ at Osaka Daigaku Ningen on June 24, 2015
tionships to body measurements of commercial strains of turkeys. Poultry Sci. 48: 1598-1603. Oakberg, E. F., 1951a. Genetic differences in quantitative histology of the adrenal, organ weights, and inter-organ correlations in White Leghorn chickens. Growth, 15: 57-78. Oakberg, E. F., 1951b. Influence of genetic constitution on growth of lymphoid tissue in liver and pancreas of White Leghorn chickens, and correlation of lymphoid tissues with weights of some visceral and endocrine organs. Growth, 15: 79-100. Oakberg, E. F., and A. M. Lucas, 1949. Variations in body weight and organ: body weight ratios of inbred lines of White Leghorn chickens in relation to mortality, especially from lymphomatosis. Growth, 13: 319-337. Roberson, R. H., and D. W. Francis, 1965. Anatomical development of White Chinese geese.
615
METHIONINE, VALINE AND NEWCASTLE
in turn may influence the gamma globulin production. In order to separate the two processes, the effect of methionine and valine on antibody production in chicks was studied using formalin killed Bl strain of Newcastle disease virus (NDV) as antigen. PROCEDURE
TABLE 1.—Basal diet Amino acid mix1 2
Salts X Vitamin mix3 Anti-acid4 Choline chloride Alpha tocopheryl acetate, 10 mg./gm. Cellulose NaHC0 3 Corn oil Dextrin to 1 kg.
Variable 60.0 5.0 10.0 2.0 1.0 30.0 10.0 90.0
Downloaded from http://ps.oxfordjournals.org/ at Osaka Daigaku Ningen on June 24, 2015
Day-old chicks, the progeny of New + Hampshire males and Single Comb White 1 Leghorn females were used in all the experAmino acid mix supplied the following in gm./ iments. All chicks were wing banded, kg. of diet: L-arginine, 20; L-glutamic acid, 70; glycine, 20; L-histidine, 6; L-isoleucine, 10; L-leucine, weighed individually and maintained in 20; L-lysine, 16; L-phenylalanine, 10; L-tryptophan, electrically heated batteries with raised 3; L-tyrosine, 9; L-valine, 11 or L-methionine, 8; L-threonine, 9. 2 wire floors. Ten chicks were used per treatSalts X supplied the following in mg./kg. of diet: CaCOs, 15,000; Ca 3 (P0 4 ) 2 , 13,950; KH 2 PO„, ment. The experimental diets and water 12,000; Na 2 HP0 4 , 7,200; NaCl, 6,000; MgS0 4 were given ad libitum from the first day af- •7H 2 0, 5,000; KC1, 3,000; MnS0 4 -2H 2 0, 420; FeC 6 H 5 0 7 , 400; ZnCOs, 200; Kl, 52; CuS0 4 , 20; ter hatching. The individual body weights NaBr, 10; NaMo0 4 , 10. 3 and group feed consumption were recorded Vitamin mix supplied in mg./kg. of diet: thi100; niacin, 100; riboflavin, 16; calcium panat the end of each week. Each experiment amine, tothenate, 20; vitamin Bi2, .02; pyridoxine HC1, 6; was terminated at the end of 18 days. biotin, .6; folic acid, 4; inositol, 100; menadione, 5; A (250,000 I.U./gm.), 40; vitamin D 3 , When the chicks were four days of age, vitamin (200,000 I.U./gm.), 5. 4 Anti-acid 3 parts magnesium trisilicate and 1 they were infected by instilling one drop of 1CT3 solution of a stock solution of Bl part aluminum hydroxide. strain of NDV in each nostril or by injecting O.S ml. per bird of 24-hour formalin in- maintained on practical diet throughout activated NDV per day into breast muscles each experiment to serve as a negative confor five days. Two weeks later (day 18), trol. the blood was obtained from the chicks by The data on weight of chicks and HI tiheart puncture and serum was collected in ter were statistically analyzed by the use of separate sterile vials. The Beta procedure Duncan's multiple range test (Steel and of hemagglutination inhibition test (Poul- Torrie, 1960). try Biologies, National Research Council, RESULTS AND DISCUSSION 1963) was used to determine the antibody titer. In a preliminary experiment, live Bl The composition of basal diet is shown strain of NDV was exposed to both 0.2 and in Table 1. The diets for methionine or va- 0.8% formalin solutions for inactivation eiline experiments were prepared by using ei- ther for 24 hours or 8 days. The inactivather methionine or valine at the prescribed tion of the virus was tested by injecting level and adding the other amino acid in (0.1 ml. per egg) into the allantoic cavity of the amounts indicated in the table. nine-day embryonated eggs and observing In addition to the groups on amino acid mortality of embryos. The results showed diets, one group of ten chicks was fed on a that 24 hours of inactivation with 0.2% practical chick starter diet (Bhargava et formalin was sufficient to inactivate the vial., 1970) and injected with killed virus. A rus. non-infected group of ten birds was also The object of the next experiment was to
616
K. K. BHARGAVA, R. P. HANSON AND M. L. SUNDE
TABLE 2.—Effect of dose and route of inoculation with killed NDV on antibody titers Experiment 2 Group
Inactivated Virus
Route
Mean H I Titer 1 (18 days)
Undiluted 10"' Undiluted 10"1
I/M I/M I/P I/P
1.20 0.50 1.20 0.33
TABLE 3.—Effect of dose and time of inoculation with killed NDV on antibody liters Experiment 3 Group #
Inactivated Virus
Day of Bleeding
1 2 3 4
Undiluted Undiluted IO-1 10"1
18 23 18 23
, H T1 Ti il ttPe ,ri M M ee a nn H 5. SO"2 5.66* 3.50b 3.16 b
1 Values are expressed as log2 of the reciprocal of last dilution. 2 Numbers with different superscript letters are significantly different (P < .05) from other numbers in the same column.
Downloaded from http://ps.oxfordjournals.org/ at Osaka Daigaku Ningen on June 24, 2015
determine the dose and route for inoculating chicks [fed practical chick starter diet (Bhargava et al, 1970)] with the inactivated virus. Two groups of six chicks each were injected intramuscularly (0.5 ml. per bird) both with undiluted and 10_1 dilution of a stock solution of 24 hours inactivated NDV for five days. The other two groups of six chicks each received intraperitoneal injections of the same dose of killed virus. Table 2 shows that the antibody response obtained with undiluted virus given intramuscularly was higher than the diluted virus (10_1) injected intraperitoneally. Since the antibody titers were very low, it was decided to hyperimmunize the chicks in subsequent experiments by injecting killed virus for five consecutive days to reach titers similar to those obtained in previous experiments. A third experiment was designed to determine the day on which the birds fed practical diet (Bhargava et al., 1970) should be bled (18th or 23rd day) in the subsequent experiments because they received the antigen for five consecutive days. The results of antibody titers in Table 3 show that varying the day of bleeding from the 18th to 23rd day did not make any significant difference in the antibody titers. The antibody titers of birds injected with undiluted killed virus were significantly (P < .05) higher than those receiving lO^1 dilution of the same virus. Since there was no significant difference between the HI titers on the 18th and 23rd day, ti-
ters in subsequent experiments were measured only on the 18th day with an appreciable saving in the diet cost. Two experiments (4 and 5) were then designed with amino acid diets containing the levels of methionine or valine as shown in Tables 4 and 5, a low level, the requirement level and one greater than the requirement were selected for each amino acid. The results (Tables 4 and 5) show that with an increase in the level of methionine or valine, better growth and feed efficiency were obtained. The requirement of methionine for maximum growth response in chicks on this amino acid diet appears to be about 0.7%, which is in agreement with the results obtained by Marrett et al. (1964), Dobson et al. (1964) and Bhargava et al. (1970). As valine was increased in the diet there was an improvement in the average weight and feed efficiency. The optimum requirement for growth with valine seems to be at least 0.9% which agrees with our previous results (Bhargava et al., 1970) and is close to the recommendations of the National Research Council (1966). The results also show that L-methionine induced a greater response than D-methionine at the 0.7% level with respect to chick growth and feed utilization. Tables 4 and 5 summarize the results on
METHIONINE, VALINE AND NEWCASTLE TABLE 4.—Effects
of methionine and valine on growth, feed efficiency and antibody production in chicks Experiment 4 Killed Virus
Diet
0.4% 0.7% 0.7% 1.1%
L-meth. L-meth. D-meth. L-meth.
. , N T-/.-M ; 0 W i" ^ a ,r. . 18 days 0-18 days
Live Virus
Geometric mean HI titer" l g dayg
65. I"3 122.21" 99.9" 143.0 d
5.10 2.00 2.35 1.90
3.88" + 0.35 4 3.88" + 0.31 3.60" + 0.44 3.70" + 0.39
71. > 138.7 d 142.8 d 103.0"
4.65 1.86 1.85 2.15
3.20"+ 3.20» + 4.00" + 3.80" +
0.32 0.32 0.36 0.32
.
AV
. , N ,-, ; r . ; 0 W J- «-» „ U*. 18 days 0-18 days
Geometric mean H I titer 2 l g dayg
4.82
4.40» + 0.37 4
14'.1*
1.95
3.30 b ±0.37
71.7"
4.50
3.20"±0.57
151.8*
1.80
4.20" + 0.32
77.4*3
1
Feed consumed/gain in weight. Values are expressed as log2 of the reciprocal of last dilution. Numbers with different superscript letters are significantly different (P<.05) from other numbers obtained in the respective experiment. 4 Mean + SEM. 2 3
antibody titers in chicks infected or injected with live or killed virus. The antibody titers in experiment 4 were lower as compared to the titers observed in previous experiments, although the differences were nonsignificant (P > .05), except in chicks fed methionine diets and infected with live virus. Table 5 shows that the HI titer was found to be significantly higher (P < .05) for 0.4% L-methionine group than for groups receiving 0.7 to 1.1% L-methionine, TABLE 5.—Effects
irrespective of the killed or live virus which they received. The antibody titer was slightly higher for L-methionine than D-methionine, but the difference was not found to be significant (P < .05). The HI titer was low in chicks fed a low level (0.5%) of valine and it increased significantly (P < .05) at higher levels (1.5%) of valine in the diet. The antibody titers of chicks (infected with live virus) fed graded levels of methionine in the diet showed a significant
of methionine and valine on growth, feed efficiency and antibody production in chicks Experiment 5 Killed Virus
Diet
0.4%, 0.7% 0.7% 1.1%
L-meth. L-meth. D-meth. L-meth.
0.5% Val. 0.9% Val. 1.5% Val. Pract. diet 1
Av.wt.(g.) F/& 18 days 0-18 days 65.2» 135.4" 90.3 b 135.5" 83.6b 137.7" 138.2" 171.4 d
3
Ge0
Live Virus g T e t t r LT a n ™ titer
5.31 2.08 2.96 1.90
5.22" + 0.22" 4.22»b + 0.40 4.2O b + 0.41 4.00 b + 0.36
3.81 2.00 1.90 2.00
2.80"±0.40 3.30" b + 0.49 4.80 b + 0.35 4.00 b ±0.36
Av.wt.(g.) l g days
F/G „_„ ^
^
S ^ 1 H I titer
67.8"
5.28
5.44"±0.42
139.8"
1.96
4.00b±0.25
80.2 b
4.27
3.00" + 0.25
d
1.80
5.20 b ±0.38
164.2
1
Feed consumed/gain in weight. Values are expressed as log2 of the reciprocal of last dilution. 3 Numbers with different superscript letters are significantly different (P<.05) from other numbers obtained in the respective experiment. 4 Mean + SEM. 2
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0.5% Val. 0.9% Val. 1.5% Val. Pract. diet
.
AV
617
618
K. K. BHAEGAVA, R. P. HANSON AND M. L. SUNDE
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(P < .05) difference in both experiments, onine decreases. and with valine the differences were signifiThe low or high antibody titers in chicks cant in experiment 5 and approached sig- fed low or high levels of valine in the diet nificance in experiment 4. These results are parallel to the observations of Koyanagree rather well with the results obtained agi and Ishiguro (1966) and Gershoff et al. in previous experiments (Bhargava et al., (1968) on amounts of gamma globulin syn1970). The chicks immunized with forma- thesized in rats fed diets containing low or lin killed virus and fed graded levels of me- high levels of tryptophan and phenylalathionine and valine showed antibody re- nine. Our results can also be explained on sponse which was similar, but slightly the basis of the fact that chicken gamma lower than the titers obtained in chicks in- globulin has a high valine content (Tenenfected with live virus. These results show house and Deutsch, 1966). that the different levels of methionine or SUMMARY valine in the diet appear to have a major influence upon antibody production and Day-old chicks were fed practical or possibly a minor one on the virus replica- amino acid diets. The effects of methionine tion. or valine deficiency or excess on antibody The results on antibody titers in chicks formation were studied in an otherwise fed methionine diets reported here using complete diet. At the end of day four, the killed or live Newcastle disease virus are chicks were injected or infected with forin agreement with the observations made malin killed or live Bl strain of Newcastle by Gill and Gershoff (1967) in monkeys, disease virus. A single dose (0.5 ml. per Gershoff et al. (1968) in rats. Ishiguro and bird) of antigen produced very low antiKoyanagi (1968) have also obtained simi- body titers, but with five injections of the lar data in studies on the effects of dietary same antigen, antibody titers increased methionine on production of gamma globu- considerably. The undiluted preparation of killed virus produced a highly significant lin in rats. The high antibody titers of chicks fed improvement in antibody titers as com_1 low levels of methionine in the diet may be pared to 10 dilution of the same virus. related to the fact that a low methionine The antibody titer was higher in chicks fed intake might have decreased the albumin 0.4% L-methionine than in chicks receiving content, but not the gamma globulin con- 0.7 and 1.1% L-methionine, irrespective of tent of serum as suggested by Ishiguro and whether the virus was live or killed. The Koyanagi (1968). In addition, the dietary growth rate, and feed efficiency improved need for methionine in antibody production with an increase in the level of methionine appears to be very low. This is in agree- in the diet. L-methionine induced a better ment with the observations of Tenenhouse response with respect to growth, feed utiliand Deutsch (1966) in which they re- zation and antibody production than D-meported that methionine is a very minor thionine. At a low level (0.5%) of valine in amino acid component of chicken gamma the diet, the antibody titer was low, but at globulin. However, the change in albumin- a high level (1.5%) it was high in chicks gamma globulin ratio and the fact that me- receiving either killed or live virus. An imthionine is a very minor constituent of provement in growth, feed efficiency, and gamma globulin do not adequately explain antibody production was observed with an the observation that antibody titers for increase in the level of dietary valine. The NDV increase as the dietary level of methi- different levels of methionine or valine in
METHIONINE, VALINE AND NEWCASTLE
the diet appear to have a major influence upon antibody production and possibly a minor influence on the virus replication. The dietary need of methionine and valine for antibody production appears to be very low and high respectively in comparison to the requirements for optimum growth. REFERENCES
Ishiguro, K., and T. Koyanagi, 1968. Effects of dietary tryptophan and methionine on protein in the serum and liver of rats. Tohoku. J. Exp. Med. 96: 345-348. Koyanagi, T., and K. Ishiguro, 1966. Dietary tryptophan and blood gamma globulin. Tohoku. J. Exp. Med. 90: 245-248. Marrett, L., H. R. Bird and M. L. Sunde, 1964. The effects of different isomers of methionine on growth of chicks fed amino acid diets. Poultry Sci. 4 3 : 1113-1118. National Research Council, 1963. Methods for the examination of poultry biologies. National Academy of Sciences—National Research Council, Washington, D.C., Publ. 70S. National Research Council, 1966. Nutrient requirements of poultry. National Academy of Sciences —National Research Council, Washington, D.C., Publ. 1345. Steel, R. G. D., and J. H. Torrie, 1960. Principles and Procedures of Statistics. McGraw-Hill Book Company, Inc., New York, p. 112. Tenenhouse, H. S., and H. F. Deutsch, 1966. Some physical-chemical properties of chicken gamma globulins and their pepsin and papain digestion products. Immunochemistry, 3 : 11-20.
Effects of Bird Density, Nutrient Density and Perches on the Performance of Caged White Leghorn Layers* R . W . DORMINEY AND G. H . ARSCOTT Department oj Poultry Science, Oregon State University, Corvallis, Oregon 97331 (Received for publication September 21, 1970)
studies have compared cage S EVERAL size and bird density with 1 to S hens per cage (Lowe and Heywang, 1964; Wilson et al, 1964, 1967; Blount, 1965; Moore et al, 1965; Owings et al, 1967; Ruggles et al, 1967; Champion and Zindel, 1968; Shirley, 1969; Ostrander and Young, 1969). Bramhall et al. (1966) and Quisenberry (1968) summarized several studies and concluded that small cages with 2 to S hens per cage providing 350 to 465 square centimeters (sq. cm.) per hen are * Oregon Agricultural Experiment Technical Paper No. 2946.
Station,
usually more suitable than larger cages providing similar space per bird. Most studies show that 1 hen per cage usually performs best, but beyond that the results vary. There is often a decline, either significant or non-significant, in egg production and an increase in mortality as the space per bird decreases. Wilson et al. (1964, 1967) found only part of the increase in mortality to be due to increased cannibalism. Body weight often decreases as the bird density increases, while egg size is usually not affected. Feed consumption per henday was no different with 1, 2, 3 or 4 heavy type layers in 30.5 X 45.7 cm. cages (Rug-
Downloaded from http://ps.oxfordjournals.org/ at Osaka Daigaku Ningen on June 24, 2015
Bhargava, K. K., R. P. Hanson and M. L. Sunde, 1970. Effects of methionine and valine on antibody production in chicks infected with Newcastle disease virus. J. Nutr. 100: 241-248. Dobson, D. C , J. O. Anderson and R. T. Warnick, 1964. A determination of the essential amino acid proportions needed to allow rapid growth in chicks. J. Nutr. 82: 67-75. Gershoff, S. N., T. J. Gill, III, S. J. Simonian and A. I. Steinberg, 1968. Some effects of amino acid deficiencies on antibody formation in the rat. J. Nutr. 9S: 184-189. Gill, T. J., I l l , and S. N. Gershoff, 1967. The effects of methionine and ethionine on antibody formation in primates. J. Immunol. 99 : 883-893.
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