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The Effect of Sulfaquinoxaline in Different Ration Formulations on Growth and Feed Efficiency of Coccidia-Free Chicks*1
EFFECT OF SULFAQUINOXALINE ON GROWTH ACKNOWLEDGMENTS
The authors are indebted to Merck and Company, Inc., Rahway, New Jersey, for providing crystalline vitamins, and to the Cerophyl Laboratories, Inc., Kansas City, Missouri, for protamone. REFERENCES
shires. 1. Requirement of laying pullets. Poultry Sci. 31:595-603. Norris, L. C , G. F. Heuser and R. F. Miller, 1950. The amount of vitamin B12 needed by chickens. Cornell Feed Service, November 10-12. Olcese, O., and J. R. Couch, 1950. Effect of injecting vitamin Bi2 into eggs from hens fed a diet low in vitamin B12. Poultry Sci. 29: 612-614. Olcese, 0 „ J. R. Couch and C. M. Lyman, 1940. Vitamin Bi2 concentrates in the nutrition of the mature domestic fowl. J. Nutrition, 41: 73-87. Peeler, H. T., R. F. Miller, C. W. Carlson, L. C. Norris and G. F. Heuser, 1951. Studies of the effect of vitamin Bi2 on hatchability. Poultry Sci. 30: 11-17. Stephenson, E. L., and B. B. Clower, 1952. The influence of an unidentified factor found in fish solubles on hatchability. Poultry Sci. 31: 936937.
The Effect of. Sulfaquinoxaline in Different Ration Formulations on Growth and Feed Efficiency of Coccidia-Free Chicks* 1 J. K. BLETNER, 2 T. B. CLARK, 2 C. E. WEAKLEY, JR., 3 AND A. H. VANLANDINGHAM4 West Virginia Agricultural Experiment Station, Morgantown . (Received for publication January 19. 1953)
T
H E effect of t h e r a p e u t i c a n d p r o p h y l a c t i c levels of s u l f a q u i n o x a l i n e o n t h e g r o w t h a n d feed efficiency of c h i c k s h a s b e e n r e p o r t e d for m a n y g r o w t h t r i a l s in which t h e chicks were either inoculated w i t h s p o r u l a t e d o o c y s t s or e x p o s e d t o n a t u r a l coccidial infections. I n only a r e l a t i v e l y few g r o w t h s t u d i e s h a v e c o c cidia-free c h i c k s b e e n u s e d , e v e n t h o u g h * Published with the approval of the Director of the West Virginia Agricultural Experiment Station as Scientific Paper No. 465. 1 The sulfaquinoxaline was supplied by Merck & Co., Rahway, New Jersey. 2 Department of Animal Husbandry, Poultry Husbandry Section. 3 Department of Agricultural Biochemistry. 4 Assistant Director, formerly Department of Agricultural Biochemistr}'.
the use of such chicks would have permitted much more critical studies of the effect of the drug on growth and feed efficiency. Using coccidia-free chicks Singsen et al. (1948) obtained larger but nonsignificant gains when a high-energy diet containing 0.01 percent sulfaquinoxaline was fed continuously for seven weeks. Similar results were reported by Koutz (1948). Larger amounts of sulfaquinoxaline have produced inconsistent results. Singsen el al. (1948) obtained a "definite reduction" in the growth of coccidia-free chicks fed a diet containing 0.03 percent sulfaquinoxaline. Their male chicks were apparently unaffected by this level of the drug. Cuckler and Malanga (1947) fed a ration
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Couch, J. R., O. Olcese, B. G. Sanders and J. V. Halick, 1950. Vitamin Bi2, APF concentrates, dried whey, fish solubles and liver fraction " L " in the nutrition of the mature fowl. J. Nutrition, 42: 473-486. Johnson, E. L., 1951. Vitamin Bi2 requirements of hens as affected by choline and antibiotic additions to the diet. Poultry Sci. 30: 919. Milligan, J. L., G. H. Arscott and G. F . Combs, 1952. Vitamin B12 requirement of New Hamp-
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J. K. BLETNER ET AL.
EXPERIMENTAL PROCEDURE
Day-old New Hampshire cockerel chicks were grown to 2 and 3 weeks of age on a non-medicated commercially mixed chick starter. At least 35 percent more chicks were started than could be used in the experiment. This permitted culling of the largest and smallest chicks. Only those chicks whose weights were closest to the mean were used. The chicks were assigned to lots on a weight basis. Every lot had approximately the same number of the larger and the smaller of the selected chicks. The average weight of all lots was the same at the beginning of the experimental period. Sixteen lots of 10 chicks
each were used in each series of trials. The chicks were maintained on the experimental ration for a 28-day period. Electrically heated battery brooders having raised wire floors were used for both the preliminary and experimental periods. The 16 battery units were placed four to a stand, and the stands were located so that the feed troughs of each unit were lighted equally by fluorescent lamps. Care was taken to keep any shadows from darkening the feed troughs. Each unit was on a shelf by itself. The shelves were spaced to allow approximately one and one-half inches between the top of a battery and the shelf above it. The arrangement greatly reduced the possibility of any battery in the stand receiving any heat from the unit either below or above it and made possible the recovery of feed thrown out of the feed trough by the chicks. Every effort was made to keep the battery room temperature uniform, but since all ventilating and heating equipment was adjusted manually the room temperature varied somewhat according to outside weather conditions. Circulating and exhaust fans were used in an attempt to equalize the temperature within the room. The temperature at the level of the bottom battery was within two degrees centigrade of that at the level of the top battery. Four different ration formulations were tested. Each ration formulation differed widely from the others in ingredients, percentage of ingredients used, or in their approximate chemical analysis. They were designated as Rations 1, 2, 3, and 4. Rations of the same formula but mixed at different times and from different lots of feedstuffs were differentiated by using a letter with the ration number, as la and lb. Rations 1 and 4 were similar to the commercial broiler rations in use prior to
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containing 0.05 percent of the drug for 26 days and obtained a slightly higher percent weight gain for chicks fed the medicated feed. Studies of the effect of sulfaquinoxaline on the feed efficiency of coccidia-free chicks have not been conclusive. Koutz (1948) observed but little difference in the amount of feed consumed by coccidiafree chicks fed 0.0125 percent sulfaquinoxaline continuously, 0.05 percent sulfaquinoxaline intermittently, and no sulfaquinoxaline. Singsen et al. (1948) noted a tendency for sulfaquinoxaline to increase the efficiency of feed utilization slightly. Since the coccidiostatic activity of sulfaquinoxaline can be inhibited by increasing the para-aminobenzoic acid content of the ration (Cuckler and Malanga, 1947), it appeared reasonable to question the effect of ration formulation on the response of chicks receiving sulfaquinoxaline. The purpose of this experiment was to study the growth and feed efficiency of coccidia-free chicks as affected by therapeutic and prophylactic levels of sulfaquinoxaline in different ration formulations when the drug was fed continuously for four weeks.
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EFFECT OF SULFAQUINOXALINE ON GROWTH TABLE 1.—Percentage composition of experimental rations Ration number Ingredients
3
4
percent
percent
percent
percent
5.0 10.0 43.45 7.5
10.0 46.2 5.0
—
—
5.0 13.75 3.75 4.5 3.0 r
Total
2.0 20.0 2.5 5.0 2.5 1.0
—.
1.0
—
1.0
0.1
—
—
1.5
1.0
— —•
— —
0.5 0.0125 0.0375
0.3 0.025 0.05
— — —
0.1
— — —
1.0 2.0 1.0 0.025 1.5 1.0
—. —
3.0
—
0.5
—
1.0 0.5 0.025 0.03 0.07
—
—
0.9 grams
— 100.0250
100.1250
—
0.5 0.0125 0.0375 1
•— — — 100.0250
22.55
21.43
20.06
(la) 21.13 (lb) 20.15
23.18
(3a) 20.82 (3b) 21.41
20.31
5.09
4.44
3.39
5.51
3.58 3.38
6.23
(3a) 28.42 (3b) 26.24
33.44
Determined fiber, percent
(la) (lb)
5.80 5.53
5.44
Dry excreta, (approximate) percent
(la) 33.48 (lb) 33.67
34.71
(3a) (3b)
'
5.0
20.27
Calculated fiber, percent
1
•— —
3.75 21.25 2.5 3.75 5.0
—
5.0 5.0 36.7 7.5 7.5 5.0 21.25
62.0
—
1.0
100.0000
Calculated protein, percent Determined protein, percent
2
Per hundred pounds of ration.
the advent of high energy rations. Ration 2 contained somewhat more crude protein and less of certain fibrous feedstuffs than Ration 1. Ration 3 was typical of the early high energy broiler rations. The chick starter rations used during the preliminary periods and Rations 1, 2, and 4 were commercially mixed feeds for which the formulae were known. Ration 3 was mixed in this laboratory. The formulae of the experimental rations together with certain analyses are given in Table 1. A highly efficient experimental design
that would permit recognition of small differences was selected. Assignment of the rations to the batteries was made by using a standard form for a 4 X 4 Latin Square Layout. This limited the number of different treatments to four per series. A ration or treatment being tested in any series was fed to four lots of chicks, one located in each battery stand and on each of the battery deck levels. This distribution made it possible to determine and remove the effect of deck level and battery stand. Previous work in this labora-
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Wheat bran Wheat flour middlings Yellow corn Ground oats Ground barley Alfalfa meal Soybean oil meal Corn gluten meal Meat scrap Fish meal Liver meal Dried whey Cane molasses Riboflavin supplement Animal protein factor supplement Ground limestone Rock phosphate-defluorinated Bone meal Iodized salt Manganese sulphate D-activated animal sterol Choline chloride Vitamin A feeding oil Niacin
1
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J. K. BLETNER ET AL.
TABLE 2.—Summary of the body weight gains (Average four lots) Percent Sulfaquinoxaline Series 1 Ration Series 2 Ration Ration Series 31 Ration Ration Series 4 Ration Ration
0.0
0.00625
0.0125
0.05
grams
grams
grams
grams
la
447
439
430
438
lb 2
496 505
— 497
— 512
RESULTS AND DISCUSSION
lb 3a
421 441
—
416
—
3b 4
425 424
— . —
407 399
The results of the growth studies are summarized in Tables 2 and 3. When 0.00625 percent sulfaquinoxaline was added to Ration la or 0.0125 percent to Rations la, 2, and 3, gains in body weight were not affected. The addition of 0.05 percent sulfaquinoxaline to Ration la in Series 1, Ration 2 in Series 2, and to Rations 3b and 4 in Series 4 did not significantly affect the gains in body weight. However, in Series 3 the addition of 0.05 percent sulfaquinoxaline to Ration 3a significantly retarded the gains in body weight. The difference in the> response of the chicks in Series 3 and 4 to the addition of 0.05 percent sulfaquinoxaline to Rations 3a and 3b is difficult to explain on a ration difference basis. Rations 3a and 3b were mixed according to the same formula but were mixed at different times. Many of the feedstuffs used in these two mixes were from the same shipments. It is realized that the 0.05 percent level of sulfaquinoxaline when fed for 28 days may approach closely the toxic level for this drug. This level of sulfaquinoxaline is four times the amount commonly recommended for use as a prophylaxis in combating avian coccidiosis. The fact that
— . — — — — —
460
Approximate least significant difference: 5% level—23 grams 1% level—30 grams
tory indicated the necessity for such an experimental design (Clark et al., 1949). Each chick was weighed weekly during the experimental period. All weights were taken to the nearest gram. The weight gains for all series, except Series 2, were determined from weights taken following about a 12-hour fasting period. Group feed consumption was determined weekly. In order to compare the effect of ration on dry matter excreted, the excreta were collected, dried, weighed, and expressed as a percentage of the feed consumed (Table 1). Members of the Animal Pathology Section made fecal examinations at the start and at the end of the experimental period to determine if the chicks carried any coccidial infection. All examinations were negative for coccidia oocysts. In Series 1, Ration la was tested with 0.0, 0.00625, 0.0125, and 0.05 percent sulfaquinoxaline. In series 2 and 3, Ration lb was used as a control. Ration 2 was
TABLE 3.-—Analysis of variance of gain in body vveight
Variance due to Between rations Between battery stands Between battery deck level Pooled error
2
1
Series D.F. 3 3 3 149
Significant at the 1% level.
Mean Squares 1,762 364 4,972 2,772
D.F. 3 3 3 149
Mean Squares 2,187 3,375 2,439 4,167
4
3 D.F. 3 3 3 150
Mean Squares 16,100* 721 4,133 2,643
D.F. 3 3 3 149
Mean Squares 6,697 11,958* 1,402 2,689
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1
tested in Series 2 and Ration 3a in Series 3. Each was fed with 0.0, 0.0125, and 0.05 percent sulfaquinoxaline. In Series 4, Rations 3b and 4 were tested with 0.0 and 0.05 percent sulfaquinoxaline.
EFFECT OF SULFAQUINOXALINK ON GROWTH
TABLE
5.— Analysis
Series Variance due to Between rations Between battery stands Between battery deck levels Remainder
* Significant at the 1% level.
D.F. 3 3 3 6
TABLE 4.—Summary of the feed-gain ratio
(Average four lots) Percent Sulfaquinoxaline
0.0
0.00625
0.0125
0.05
(Grams feed per gram gain) Series 1 Ration Series 2 Ration Ration Series 3 Ration Ration Series 4 Ration Ration
la
2.7248
2.7571
2.7786
lb 2
2.9215 2.7871
— —
2.8090
—
2.7444
lb 2a
2.6503 2.5591
—
—
2.5240
—
2.5795
3b 4
2.6621 2.9483
— —
2.7222
—
—
2.6938 3.0847
— —
Approximate least significant difference: 5% level 1% level Series 2 0.0713 0.1081 Series 3 0.0574 0.0869 Series 4 0.1090 0.1651
parent. However, feed wastage is a highly variable factor affecting feed utilization data. It was observed that certain lots of chicks wasted more feed than did other lots on the same ration. With proper management it was possible to keep such losses at a minimum. The water troughs often contained large amounts of feed and there was very little that could be done to reduce this error. What effect this may have had on these results is not known. In these experiments there was no tendency for sulfaquinoxaline to increase feed utilization as was reported by Singsen et al. (1948). In general, the more rapidly growing chicks required less feed per gram of gain than was required by the slower growing chicks. The feed utilization also varied within any series with the crude fiber and crude protein content of the rations. The feed-gain ratios for the high fiber rations were higher than those
of variance of feed-gain ratios 1
2
3
4
Mean Squares 0.0029 0.0024 0.0096 0.00295
Mean Squares 0.0229* 0.0217* 0.0030 0.0017
Mean Squares 0.0113* 0.0027 0.0034 0.0011
Mean Squares 0.1659* 0.0104 0.0047 0.00397
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this high level of sulfaquinoxaline fed continuously for 28 days significantly retarded growth only when fed with Ration 3, and then only in one of two trials, further indicates the relative safety with which this drug may be used. Further study of the effect of other ration formulations on the response of chicks to this drug might yield valuable information. In none of the trials reported here was a significant increase in the growth of coccidia-free chicks obtained. It may be inferred that the increased growth reported by other workers studying floorbrooded chicks fed prophylactic levels of sulfaquinoxaline was probably not due to any growth stimulating property of the drug (Jungherr and Winn, 1949; Lucas et al, 1949; Hart et al, 1949; Grumbles and Delaplane, 1947, 1948; Grumbles, Delaplane and Higgins, 1948 a, b). The results of the feed efficiency studies are presented in Tables 4 and 5. The feedgain ratios (grams of feed required per gram of gain) for the rations containing sulfaquinoxaline differ significantly from those ratios for the rations containing none of the drug in Series 4 only. In that series Ration 4 when fed with 0.05 percent sulfaquinoxaline was used less efficiently than the same ration without the drug. The difference between these ratios is significant at the 5 percent level. Since Rations la and 4 were somewhat similar, it is considered that the results obtained in Series 1 and 4 are not in agreement. The reason for this difference is not ap-
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J. K. BLETNER ET AL.
TABLE 6.—Body weight gains according to battery stands-series 4 Stand
Gain
1 2 3 4
grams 435 393 415 412
Approx. L.S.D. 5% level—23 grams 1% level—'30 grams
No similar differences in the body weight studies were found in the other series, and in none of the series were significant differences found between battery deck levels. The fact that unexplained locational difference did appear in one of the four series is important. This indicates that one or two experiments may not be sufficient to determine whether or not unexplained locational difference may be present in a laboratory and even suggests that such differences are likely to occur at any time in any laboratory. Locational differences have been observed by Hill et al. (1944), Carrick and Roberts (1947, 1948), and by Clark et al. (1949). The effect of locational differences was removed in the analysis of these data by the use of the 4 X 4 Latin Square Layout. When the feed-gain ratios were analyzed (Table 5), a highly significant difference between battery stands was obtained for Series 2. The feed-gain ratio for
Stand 3 was significantly larger than that for any of the other three stands. This may be another example of locational difference, but it is believed more likely to be the result of uneven distribution of female chicks. The sexing error for this series was large, there being 36 females among the 160 experimental chicks. Fifteen of these female chicks were in this battery stand. The effect of this added variability due to the unequal distribution of sexes following a large sexing error was removed in this experimental design along with the stand differences. The data used for determining the weight gains of the chicks in Series 1, 3, and 4 were obtained from body weights taken after the chicks had been fasted about 12 hours. The chicks in Series 2 were weighed without a fasting period. To test the effect of fasting as a means of reducing the variability the chicks in Series 3 and 4 were weighed before and after a fasting period of about 12 hours during which time both feed and water were withheld. The coefficients of variation of the gains determined from the non-fasted and fasted weights were 11.94 and 11.93 percent, respectively, in Series 3 and 12.42 and 12.52 percent, respectively, in Series 4. Thus, in Series 3 and 4 the use of a 12-hour fasting period, prior to weighing the experimental chicks, did not affect the coefficient of variation of the weight gains and neither did it have any effect on the conclusions. Fasting did reduce the variability of the data but it also reduced the average weight gain proportionately. The mortality during the 4-week experimental period was very low in all series. Only one chick died in each of Series 1, 2, and 4, and none in Series 3. SUMMARY
Coccidia-free New Hampshire cockerel
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for the lower fiber or higher protein rations in the same series. These differences were highly significant. Unexplained locational differences did appear when the gains in body weight were analyzed for Series 4 (Table 3). The difference between the average gain made by the chicks in Battery Stands 1 and 2 was highly significant, and between Battery Stands 1 and 4 the difference was significant at the 5 percent level (Table 6).
NEWS AND NOTES
REFERENCES Carrick, C. W., and R. E. Roberts, 1947. Studies on feed efficiency. I. The relative efficiency of corn and wheat products in chick rations. Poultry Sci. 26:111-117. Carrick, C. W., and R. E. Roberts, 1948. Studies on feed efficiency. I I . The relative efficiency of corn and oats in chick rations. Poultry Sci. 27: 213218. Clark, T. B., A. H. VanLandingham and C. E. Weakley, Jr., 1949. Soybean oil meals in poultry rations. I I . Soybean oil meals alone and in combination with meat scrap or corn gluten meal. Poultry Sci. 28: 521-529.
Cuckler, A. C , and C. Malanga, 1947. A summary of experiments with sulfaquinoxaline for cecal coccidiosis. "Conference on Sulfaquinoxaline," Merck & Co., Inc. 71-78. Grumbles, L. C , and J. P . Delaplane, 1947. Sulfaquinoxaline in the prevention of Eimeria necatrix infection in chickens. Proc. 51st Ann. Mtg. U. S. Livestock Sanitary Assoc. 285-289. Grumbles, L. C , and J. P. Delaplane, 1948. Relative activity of sulfamethazine and sulfaquinoxaline against Eimeria tennella infections in young chickens. Am. J. Vet. Res. 9:306-309. . Grumbles, L. C , J. P. Delaplane and T. C. Higgins, 1948a. Prophylactic and therapeutic use of sulfaquinoxaline against coccidia of chickens {Eimeria tennella and Eimeria necatrix) under field conditions. Poultry Sci. 27: 411-419. Grumbles, L. C , J. P . Delaplane and T. C. Higgins, 1948b. Continuous feeding of low concentrations of sulfaquinoxaline for the control of coccidiosis in poultry. Poultry Sci. 27: 605-608. Hart, C. P., W. H. Wiley, J. P. Delaplane, L. C. Grumbles and T. C. Higgins, 1949. Medication versus sanitation in control of coccidiosis. Poultry Sci. 28: 686-690. Hill, D. C , S. J. Slinger and I. Motzok, 1944. Variation in the growth-rate of experimental birds. Poultry Sci. 23: 461-463. Jungherr, E. L., and J. D. Winn, 1949. Continuous low level sulfaquinoxaline feeding in the practical control of coccidiosis in broilers. Ann. New York Acad. Sci. 52: 563-570. Koutz, F . R., 1948. Immunity studies in avian cecal coccidiosis. I. The value of drugs to establish immunity in young chickens. Am. J. Vet. Res. 9: 388-395. Lucas, W. C , A. E. Tomhave and K. C. Seeger, 1949. The effect of nitrophenide, sulfaquinoxaline, sulfamethazine and sulfaguanidine in the control of caecal coccidiosis. Delaware Agr. Expt. Sta. Bull. 280. Singsen, E. P., H. M. Scott and L. D. Matterson, 1948. The effect of sulfaquinoxaline on growth rate and feed efficiency of chicks. Poultry Sci. 27: 627-628.
NEWS AND NOTES (Continued from page 729) lege, representing the American Society of Animal vestigations of the Animal Husbandry Division of Production; and K. L. Turk, Cornell University, rep- the Bureau of Animal Industry. He succeeds H. R. Bird who resigned early this year to head poultry resenting the American Dairy Science Association. nutrition research at the University of Wisconsin. U.S.D.A. NOTES A native of Kansas, Dr. Brant graduated from A. W. Brant has been named Head of Poultry In- Kansas State College in 1940, majoring in poultry (Continued on page 742)
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chicks were used in four series of growth trials to study the effect of sulfaquinoxaline on growth and feed efficiency when the drug was fed at varying levels in different ration formulations. The drug was fed for a 4-week period beginning when the chicks were two or three weeks of age. Two of the levels of sulfaquinoxaline tested, 0.00625 and 0.0125 percent, with different rations did not affect either growth or feed utilization. The therapeutic level of the drug, 0.05 percent, fed continuously instead of intermittently, as recommended, gave slightly inconsistent results indicating that continuous feeding at this level may be slightly toxic in certain rations. Observations on experimental techniques emphasize the importance of sufficient replicate lots and of experimental design in minimizing the effect of variability due to unexplained locational differences. Fasting experimental chicks for 12 hours before weighing did not reduce the coefficient of variability of the weight gains or alter the conclusions.
739
The authors are indebted to Merck and Company, Inc., Rahway, New Jersey, for providing crystalline vitamins, and to the Cerophyl Laboratories, Inc., Kansas City, Missouri, for protamone. REFERENCES
shires. 1. Requirement of laying pullets. Poultry Sci. 31:595-603. Norris, L. C , G. F. Heuser and R. F. Miller, 1950. The amount of vitamin B12 needed by chickens. Cornell Feed Service, November 10-12. Olcese, O., and J. R. Couch, 1950. Effect of injecting vitamin Bi2 into eggs from hens fed a diet low in vitamin B12. Poultry Sci. 29: 612-614. Olcese, 0 „ J. R. Couch and C. M. Lyman, 1940. Vitamin Bi2 concentrates in the nutrition of the mature domestic fowl. J. Nutrition, 41: 73-87. Peeler, H. T., R. F. Miller, C. W. Carlson, L. C. Norris and G. F. Heuser, 1951. Studies of the effect of vitamin Bi2 on hatchability. Poultry Sci. 30: 11-17. Stephenson, E. L., and B. B. Clower, 1952. The influence of an unidentified factor found in fish solubles on hatchability. Poultry Sci. 31: 936937.
The Effect of. Sulfaquinoxaline in Different Ration Formulations on Growth and Feed Efficiency of Coccidia-Free Chicks* 1 J. K. BLETNER, 2 T. B. CLARK, 2 C. E. WEAKLEY, JR., 3 AND A. H. VANLANDINGHAM4 West Virginia Agricultural Experiment Station, Morgantown . (Received for publication January 19. 1953)
T
H E effect of t h e r a p e u t i c a n d p r o p h y l a c t i c levels of s u l f a q u i n o x a l i n e o n t h e g r o w t h a n d feed efficiency of c h i c k s h a s b e e n r e p o r t e d for m a n y g r o w t h t r i a l s in which t h e chicks were either inoculated w i t h s p o r u l a t e d o o c y s t s or e x p o s e d t o n a t u r a l coccidial infections. I n only a r e l a t i v e l y few g r o w t h s t u d i e s h a v e c o c cidia-free c h i c k s b e e n u s e d , e v e n t h o u g h * Published with the approval of the Director of the West Virginia Agricultural Experiment Station as Scientific Paper No. 465. 1 The sulfaquinoxaline was supplied by Merck & Co., Rahway, New Jersey. 2 Department of Animal Husbandry, Poultry Husbandry Section. 3 Department of Agricultural Biochemistry. 4 Assistant Director, formerly Department of Agricultural Biochemistr}'.
the use of such chicks would have permitted much more critical studies of the effect of the drug on growth and feed efficiency. Using coccidia-free chicks Singsen et al. (1948) obtained larger but nonsignificant gains when a high-energy diet containing 0.01 percent sulfaquinoxaline was fed continuously for seven weeks. Similar results were reported by Koutz (1948). Larger amounts of sulfaquinoxaline have produced inconsistent results. Singsen el al. (1948) obtained a "definite reduction" in the growth of coccidia-free chicks fed a diet containing 0.03 percent sulfaquinoxaline. Their male chicks were apparently unaffected by this level of the drug. Cuckler and Malanga (1947) fed a ration
Downloaded from http://ps.oxfordjournals.org/ at University of North Dakota on May 20, 2015
Couch, J. R., O. Olcese, B. G. Sanders and J. V. Halick, 1950. Vitamin Bi2, APF concentrates, dried whey, fish solubles and liver fraction " L " in the nutrition of the mature fowl. J. Nutrition, 42: 473-486. Johnson, E. L., 1951. Vitamin Bi2 requirements of hens as affected by choline and antibiotic additions to the diet. Poultry Sci. 30: 919. Milligan, J. L., G. H. Arscott and G. F . Combs, 1952. Vitamin B12 requirement of New Hamp-
733
734
J. K. BLETNER ET AL.
EXPERIMENTAL PROCEDURE
Day-old New Hampshire cockerel chicks were grown to 2 and 3 weeks of age on a non-medicated commercially mixed chick starter. At least 35 percent more chicks were started than could be used in the experiment. This permitted culling of the largest and smallest chicks. Only those chicks whose weights were closest to the mean were used. The chicks were assigned to lots on a weight basis. Every lot had approximately the same number of the larger and the smaller of the selected chicks. The average weight of all lots was the same at the beginning of the experimental period. Sixteen lots of 10 chicks
each were used in each series of trials. The chicks were maintained on the experimental ration for a 28-day period. Electrically heated battery brooders having raised wire floors were used for both the preliminary and experimental periods. The 16 battery units were placed four to a stand, and the stands were located so that the feed troughs of each unit were lighted equally by fluorescent lamps. Care was taken to keep any shadows from darkening the feed troughs. Each unit was on a shelf by itself. The shelves were spaced to allow approximately one and one-half inches between the top of a battery and the shelf above it. The arrangement greatly reduced the possibility of any battery in the stand receiving any heat from the unit either below or above it and made possible the recovery of feed thrown out of the feed trough by the chicks. Every effort was made to keep the battery room temperature uniform, but since all ventilating and heating equipment was adjusted manually the room temperature varied somewhat according to outside weather conditions. Circulating and exhaust fans were used in an attempt to equalize the temperature within the room. The temperature at the level of the bottom battery was within two degrees centigrade of that at the level of the top battery. Four different ration formulations were tested. Each ration formulation differed widely from the others in ingredients, percentage of ingredients used, or in their approximate chemical analysis. They were designated as Rations 1, 2, 3, and 4. Rations of the same formula but mixed at different times and from different lots of feedstuffs were differentiated by using a letter with the ration number, as la and lb. Rations 1 and 4 were similar to the commercial broiler rations in use prior to
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containing 0.05 percent of the drug for 26 days and obtained a slightly higher percent weight gain for chicks fed the medicated feed. Studies of the effect of sulfaquinoxaline on the feed efficiency of coccidia-free chicks have not been conclusive. Koutz (1948) observed but little difference in the amount of feed consumed by coccidiafree chicks fed 0.0125 percent sulfaquinoxaline continuously, 0.05 percent sulfaquinoxaline intermittently, and no sulfaquinoxaline. Singsen et al. (1948) noted a tendency for sulfaquinoxaline to increase the efficiency of feed utilization slightly. Since the coccidiostatic activity of sulfaquinoxaline can be inhibited by increasing the para-aminobenzoic acid content of the ration (Cuckler and Malanga, 1947), it appeared reasonable to question the effect of ration formulation on the response of chicks receiving sulfaquinoxaline. The purpose of this experiment was to study the growth and feed efficiency of coccidia-free chicks as affected by therapeutic and prophylactic levels of sulfaquinoxaline in different ration formulations when the drug was fed continuously for four weeks.
735
EFFECT OF SULFAQUINOXALINE ON GROWTH TABLE 1.—Percentage composition of experimental rations Ration number Ingredients
3
4
percent
percent
percent
percent
5.0 10.0 43.45 7.5
10.0 46.2 5.0
—
—
5.0 13.75 3.75 4.5 3.0 r
Total
2.0 20.0 2.5 5.0 2.5 1.0
—.
1.0
—
1.0
0.1
—
—
1.5
1.0
— —•
— —
0.5 0.0125 0.0375
0.3 0.025 0.05
— — —
0.1
— — —
1.0 2.0 1.0 0.025 1.5 1.0
—. —
3.0
—
0.5
—
1.0 0.5 0.025 0.03 0.07
—
—
0.9 grams
— 100.0250
100.1250
—
0.5 0.0125 0.0375 1
•— — — 100.0250
22.55
21.43
20.06
(la) 21.13 (lb) 20.15
23.18
(3a) 20.82 (3b) 21.41
20.31
5.09
4.44
3.39
5.51
3.58 3.38
6.23
(3a) 28.42 (3b) 26.24
33.44
Determined fiber, percent
(la) (lb)
5.80 5.53
5.44
Dry excreta, (approximate) percent
(la) 33.48 (lb) 33.67
34.71
(3a) (3b)
'
5.0
20.27
Calculated fiber, percent
1
•— —
3.75 21.25 2.5 3.75 5.0
—
5.0 5.0 36.7 7.5 7.5 5.0 21.25
62.0
—
1.0
100.0000
Calculated protein, percent Determined protein, percent
2
Per hundred pounds of ration.
the advent of high energy rations. Ration 2 contained somewhat more crude protein and less of certain fibrous feedstuffs than Ration 1. Ration 3 was typical of the early high energy broiler rations. The chick starter rations used during the preliminary periods and Rations 1, 2, and 4 were commercially mixed feeds for which the formulae were known. Ration 3 was mixed in this laboratory. The formulae of the experimental rations together with certain analyses are given in Table 1. A highly efficient experimental design
that would permit recognition of small differences was selected. Assignment of the rations to the batteries was made by using a standard form for a 4 X 4 Latin Square Layout. This limited the number of different treatments to four per series. A ration or treatment being tested in any series was fed to four lots of chicks, one located in each battery stand and on each of the battery deck levels. This distribution made it possible to determine and remove the effect of deck level and battery stand. Previous work in this labora-
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Wheat bran Wheat flour middlings Yellow corn Ground oats Ground barley Alfalfa meal Soybean oil meal Corn gluten meal Meat scrap Fish meal Liver meal Dried whey Cane molasses Riboflavin supplement Animal protein factor supplement Ground limestone Rock phosphate-defluorinated Bone meal Iodized salt Manganese sulphate D-activated animal sterol Choline chloride Vitamin A feeding oil Niacin
1
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J. K. BLETNER ET AL.
TABLE 2.—Summary of the body weight gains (Average four lots) Percent Sulfaquinoxaline Series 1 Ration Series 2 Ration Ration Series 31 Ration Ration Series 4 Ration Ration
0.0
0.00625
0.0125
0.05
grams
grams
grams
grams
la
447
439
430
438
lb 2
496 505
— 497
— 512
RESULTS AND DISCUSSION
lb 3a
421 441
—
416
—
3b 4
425 424
— . —
407 399
The results of the growth studies are summarized in Tables 2 and 3. When 0.00625 percent sulfaquinoxaline was added to Ration la or 0.0125 percent to Rations la, 2, and 3, gains in body weight were not affected. The addition of 0.05 percent sulfaquinoxaline to Ration la in Series 1, Ration 2 in Series 2, and to Rations 3b and 4 in Series 4 did not significantly affect the gains in body weight. However, in Series 3 the addition of 0.05 percent sulfaquinoxaline to Ration 3a significantly retarded the gains in body weight. The difference in the> response of the chicks in Series 3 and 4 to the addition of 0.05 percent sulfaquinoxaline to Rations 3a and 3b is difficult to explain on a ration difference basis. Rations 3a and 3b were mixed according to the same formula but were mixed at different times. Many of the feedstuffs used in these two mixes were from the same shipments. It is realized that the 0.05 percent level of sulfaquinoxaline when fed for 28 days may approach closely the toxic level for this drug. This level of sulfaquinoxaline is four times the amount commonly recommended for use as a prophylaxis in combating avian coccidiosis. The fact that
— . — — — — —
460
Approximate least significant difference: 5% level—23 grams 1% level—30 grams
tory indicated the necessity for such an experimental design (Clark et al., 1949). Each chick was weighed weekly during the experimental period. All weights were taken to the nearest gram. The weight gains for all series, except Series 2, were determined from weights taken following about a 12-hour fasting period. Group feed consumption was determined weekly. In order to compare the effect of ration on dry matter excreted, the excreta were collected, dried, weighed, and expressed as a percentage of the feed consumed (Table 1). Members of the Animal Pathology Section made fecal examinations at the start and at the end of the experimental period to determine if the chicks carried any coccidial infection. All examinations were negative for coccidia oocysts. In Series 1, Ration la was tested with 0.0, 0.00625, 0.0125, and 0.05 percent sulfaquinoxaline. In series 2 and 3, Ration lb was used as a control. Ration 2 was
TABLE 3.-—Analysis of variance of gain in body vveight
Variance due to Between rations Between battery stands Between battery deck level Pooled error
2
1
Series D.F. 3 3 3 149
Significant at the 1% level.
Mean Squares 1,762 364 4,972 2,772
D.F. 3 3 3 149
Mean Squares 2,187 3,375 2,439 4,167
4
3 D.F. 3 3 3 150
Mean Squares 16,100* 721 4,133 2,643
D.F. 3 3 3 149
Mean Squares 6,697 11,958* 1,402 2,689
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1
tested in Series 2 and Ration 3a in Series 3. Each was fed with 0.0, 0.0125, and 0.05 percent sulfaquinoxaline. In Series 4, Rations 3b and 4 were tested with 0.0 and 0.05 percent sulfaquinoxaline.
EFFECT OF SULFAQUINOXALINK ON GROWTH
TABLE
5.— Analysis
Series Variance due to Between rations Between battery stands Between battery deck levels Remainder
* Significant at the 1% level.
D.F. 3 3 3 6
TABLE 4.—Summary of the feed-gain ratio
(Average four lots) Percent Sulfaquinoxaline
0.0
0.00625
0.0125
0.05
(Grams feed per gram gain) Series 1 Ration Series 2 Ration Ration Series 3 Ration Ration Series 4 Ration Ration
la
2.7248
2.7571
2.7786
lb 2
2.9215 2.7871
— —
2.8090
—
2.7444
lb 2a
2.6503 2.5591
—
—
2.5240
—
2.5795
3b 4
2.6621 2.9483
— —
2.7222
—
—
2.6938 3.0847
— —
Approximate least significant difference: 5% level 1% level Series 2 0.0713 0.1081 Series 3 0.0574 0.0869 Series 4 0.1090 0.1651
parent. However, feed wastage is a highly variable factor affecting feed utilization data. It was observed that certain lots of chicks wasted more feed than did other lots on the same ration. With proper management it was possible to keep such losses at a minimum. The water troughs often contained large amounts of feed and there was very little that could be done to reduce this error. What effect this may have had on these results is not known. In these experiments there was no tendency for sulfaquinoxaline to increase feed utilization as was reported by Singsen et al. (1948). In general, the more rapidly growing chicks required less feed per gram of gain than was required by the slower growing chicks. The feed utilization also varied within any series with the crude fiber and crude protein content of the rations. The feed-gain ratios for the high fiber rations were higher than those
of variance of feed-gain ratios 1
2
3
4
Mean Squares 0.0029 0.0024 0.0096 0.00295
Mean Squares 0.0229* 0.0217* 0.0030 0.0017
Mean Squares 0.0113* 0.0027 0.0034 0.0011
Mean Squares 0.1659* 0.0104 0.0047 0.00397
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this high level of sulfaquinoxaline fed continuously for 28 days significantly retarded growth only when fed with Ration 3, and then only in one of two trials, further indicates the relative safety with which this drug may be used. Further study of the effect of other ration formulations on the response of chicks to this drug might yield valuable information. In none of the trials reported here was a significant increase in the growth of coccidia-free chicks obtained. It may be inferred that the increased growth reported by other workers studying floorbrooded chicks fed prophylactic levels of sulfaquinoxaline was probably not due to any growth stimulating property of the drug (Jungherr and Winn, 1949; Lucas et al, 1949; Hart et al, 1949; Grumbles and Delaplane, 1947, 1948; Grumbles, Delaplane and Higgins, 1948 a, b). The results of the feed efficiency studies are presented in Tables 4 and 5. The feedgain ratios (grams of feed required per gram of gain) for the rations containing sulfaquinoxaline differ significantly from those ratios for the rations containing none of the drug in Series 4 only. In that series Ration 4 when fed with 0.05 percent sulfaquinoxaline was used less efficiently than the same ration without the drug. The difference between these ratios is significant at the 5 percent level. Since Rations la and 4 were somewhat similar, it is considered that the results obtained in Series 1 and 4 are not in agreement. The reason for this difference is not ap-
737
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J. K. BLETNER ET AL.
TABLE 6.—Body weight gains according to battery stands-series 4 Stand
Gain
1 2 3 4
grams 435 393 415 412
Approx. L.S.D. 5% level—23 grams 1% level—'30 grams
No similar differences in the body weight studies were found in the other series, and in none of the series were significant differences found between battery deck levels. The fact that unexplained locational difference did appear in one of the four series is important. This indicates that one or two experiments may not be sufficient to determine whether or not unexplained locational difference may be present in a laboratory and even suggests that such differences are likely to occur at any time in any laboratory. Locational differences have been observed by Hill et al. (1944), Carrick and Roberts (1947, 1948), and by Clark et al. (1949). The effect of locational differences was removed in the analysis of these data by the use of the 4 X 4 Latin Square Layout. When the feed-gain ratios were analyzed (Table 5), a highly significant difference between battery stands was obtained for Series 2. The feed-gain ratio for
Stand 3 was significantly larger than that for any of the other three stands. This may be another example of locational difference, but it is believed more likely to be the result of uneven distribution of female chicks. The sexing error for this series was large, there being 36 females among the 160 experimental chicks. Fifteen of these female chicks were in this battery stand. The effect of this added variability due to the unequal distribution of sexes following a large sexing error was removed in this experimental design along with the stand differences. The data used for determining the weight gains of the chicks in Series 1, 3, and 4 were obtained from body weights taken after the chicks had been fasted about 12 hours. The chicks in Series 2 were weighed without a fasting period. To test the effect of fasting as a means of reducing the variability the chicks in Series 3 and 4 were weighed before and after a fasting period of about 12 hours during which time both feed and water were withheld. The coefficients of variation of the gains determined from the non-fasted and fasted weights were 11.94 and 11.93 percent, respectively, in Series 3 and 12.42 and 12.52 percent, respectively, in Series 4. Thus, in Series 3 and 4 the use of a 12-hour fasting period, prior to weighing the experimental chicks, did not affect the coefficient of variation of the weight gains and neither did it have any effect on the conclusions. Fasting did reduce the variability of the data but it also reduced the average weight gain proportionately. The mortality during the 4-week experimental period was very low in all series. Only one chick died in each of Series 1, 2, and 4, and none in Series 3. SUMMARY
Coccidia-free New Hampshire cockerel
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for the lower fiber or higher protein rations in the same series. These differences were highly significant. Unexplained locational differences did appear when the gains in body weight were analyzed for Series 4 (Table 3). The difference between the average gain made by the chicks in Battery Stands 1 and 2 was highly significant, and between Battery Stands 1 and 4 the difference was significant at the 5 percent level (Table 6).
NEWS AND NOTES
REFERENCES Carrick, C. W., and R. E. Roberts, 1947. Studies on feed efficiency. I. The relative efficiency of corn and wheat products in chick rations. Poultry Sci. 26:111-117. Carrick, C. W., and R. E. Roberts, 1948. Studies on feed efficiency. I I . The relative efficiency of corn and oats in chick rations. Poultry Sci. 27: 213218. Clark, T. B., A. H. VanLandingham and C. E. Weakley, Jr., 1949. Soybean oil meals in poultry rations. I I . Soybean oil meals alone and in combination with meat scrap or corn gluten meal. Poultry Sci. 28: 521-529.
Cuckler, A. C , and C. Malanga, 1947. A summary of experiments with sulfaquinoxaline for cecal coccidiosis. "Conference on Sulfaquinoxaline," Merck & Co., Inc. 71-78. Grumbles, L. C , and J. P . Delaplane, 1947. Sulfaquinoxaline in the prevention of Eimeria necatrix infection in chickens. Proc. 51st Ann. Mtg. U. S. Livestock Sanitary Assoc. 285-289. Grumbles, L. C , and J. P. Delaplane, 1948. Relative activity of sulfamethazine and sulfaquinoxaline against Eimeria tennella infections in young chickens. Am. J. Vet. Res. 9:306-309. . Grumbles, L. C , J. P. Delaplane and T. C. Higgins, 1948a. Prophylactic and therapeutic use of sulfaquinoxaline against coccidia of chickens {Eimeria tennella and Eimeria necatrix) under field conditions. Poultry Sci. 27: 411-419. Grumbles, L. C , J. P . Delaplane and T. C. Higgins, 1948b. Continuous feeding of low concentrations of sulfaquinoxaline for the control of coccidiosis in poultry. Poultry Sci. 27: 605-608. Hart, C. P., W. H. Wiley, J. P. Delaplane, L. C. Grumbles and T. C. Higgins, 1949. Medication versus sanitation in control of coccidiosis. Poultry Sci. 28: 686-690. Hill, D. C , S. J. Slinger and I. Motzok, 1944. Variation in the growth-rate of experimental birds. Poultry Sci. 23: 461-463. Jungherr, E. L., and J. D. Winn, 1949. Continuous low level sulfaquinoxaline feeding in the practical control of coccidiosis in broilers. Ann. New York Acad. Sci. 52: 563-570. Koutz, F . R., 1948. Immunity studies in avian cecal coccidiosis. I. The value of drugs to establish immunity in young chickens. Am. J. Vet. Res. 9: 388-395. Lucas, W. C , A. E. Tomhave and K. C. Seeger, 1949. The effect of nitrophenide, sulfaquinoxaline, sulfamethazine and sulfaguanidine in the control of caecal coccidiosis. Delaware Agr. Expt. Sta. Bull. 280. Singsen, E. P., H. M. Scott and L. D. Matterson, 1948. The effect of sulfaquinoxaline on growth rate and feed efficiency of chicks. Poultry Sci. 27: 627-628.
NEWS AND NOTES (Continued from page 729) lege, representing the American Society of Animal vestigations of the Animal Husbandry Division of Production; and K. L. Turk, Cornell University, rep- the Bureau of Animal Industry. He succeeds H. R. Bird who resigned early this year to head poultry resenting the American Dairy Science Association. nutrition research at the University of Wisconsin. U.S.D.A. NOTES A native of Kansas, Dr. Brant graduated from A. W. Brant has been named Head of Poultry In- Kansas State College in 1940, majoring in poultry (Continued on page 742)
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chicks were used in four series of growth trials to study the effect of sulfaquinoxaline on growth and feed efficiency when the drug was fed at varying levels in different ration formulations. The drug was fed for a 4-week period beginning when the chicks were two or three weeks of age. Two of the levels of sulfaquinoxaline tested, 0.00625 and 0.0125 percent, with different rations did not affect either growth or feed utilization. The therapeutic level of the drug, 0.05 percent, fed continuously instead of intermittently, as recommended, gave slightly inconsistent results indicating that continuous feeding at this level may be slightly toxic in certain rations. Observations on experimental techniques emphasize the importance of sufficient replicate lots and of experimental design in minimizing the effect of variability due to unexplained locational differences. Fasting experimental chicks for 12 hours before weighing did not reduce the coefficient of variability of the weight gains or alter the conclusions.
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