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The Deposition and Clearance of Rofenaid® in Chicken and Turkey Eggs
The Deposition and Clearance of Rofenaid® in Chicken and Turkey Eggs H. J. LAURENCOT, A. SCHLOSSER AND J. L. HEMPSTEAD Chemical Research Department, Hoffmann-La Roche Inc., Nutley, New Jersey 07110 (Received for publication October 23, 1971)
POULTRY SCIENCE 51: 1181-1187, 1972
INTRODUCTION OFENAID®* is a new broad spectrum coccidiostat and antibacterial for poultry. It is a combination of sulfadimethoxine (SDM) (4-sulfanilamido-2,6-dimethoxypyrimidine) and ormetoprim (2,4-diamino-S[ 4,5-dimethoxy- 2 -methylbenzyl ] pyrimidine) in a ratio of 5:3. The anticoccidial activity of Rofenaid has already been reported in the chicken (Mitrovic et al., 1969a) and in the turkey (Mitrovic et al., 1971a), as has also the antibacterial activity in the chicken (Mitrovic et al., 1969b) and in the turkey (Mitrovic et al., 1971b). The safety and compatibility of Rofenaid have been reported in the chicken (Marusich et al., 1969) and in the turkey (Marusich et al., 1971). The present report is concerned with the deposition of Rofenaid into the eggs of laying chicken and
R
* Rofenaid is the Hoffmann-La Roche trademark for a coccidiostat and antibacterial containing sulfadimethoxine and ormetoprim.
turkey hens, and with the length of time required for developing eggs to be clear of the drug, in order to establish a reasonable clearance time for replacement pullets and young breeder hens. MATERIAL AND METHODS Rofenaid, labelled with radioactive carbon-14 either in the SDM or in the ormetoprim portion, was administered to laying chicken hens at a level of 0.02% in the feed, and to laying turkey hens at 0.01% in the feed for 14 days. Eggs were collected daily, separated into yolk, albumen and shell fractions, and assayed for radioactivity by liquid scintillation counting, using a modification of previously reported solubilizing techniques (Laurencot et al., 1971). After the radioactivity had reached a plateau, feeding of the drug was discontinued and analysis of the eggs was continued until no more radioactivity could be detected. Two two-year old Shaver Leghorn hens were used for the determination of the de-
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ABSTRACT Rofenaid® is a coccidiostat and antibacterial for poultry. The active ingredients are a combination of sulfadimethoxine and ormetoprim in a ratio of 5 to 3. The deposition and clearance of Rofenaid® in chicken and turkey eggs were studied in order to establish a clearance time for replacement pullets and young breeder hens. Rofenaid®, labelled with radioactive carbon-" either in the sulfadimethoxine or in the ormetoprim portion was administered to laying chicken hens at a level of 0.02% in the feed, and to laying turkey hens at 0.01% in the feed. Eggs were collected daily, separated into yolk, albumen and shell fractions, and assayed for radioactivity by liquid scintillation counting. When the radioactivity had reached a plateau, feeding of the drug was discontinued and an analysis of the eggs was continued until no more radioactivity could be detected. In the chicken, 0.59% of the daily intake of sulfadimethoxine and 0.83% of the daily intake of ormetoprim were transferred to the whole egg at the plateau level. The time required for the whole egg to clear to the 0.01 p.p.m. level was estimated at 9 days for sulfadimethoxine and 7 days for ormetoprim. In the turkey, 0.71% of the daily intake of sulfadimethoxine and 0.17% of the daily intake of ormetoprim were transferred to the whole egg at the plateau level. The time required for the whole egg to clear to the 0.01 p.p.m. level was estimated at 14 days for sulfadimethoxine and 12 days for ormetoprim.
1182
H. J. LAURENCOT, A. SCHLOSSER AND J. L. HEMPSTEAD
RESULTS In Table 1, the biological data for the four individual treatments are presented. The data are expressed as the average from the individual group of birds used in each treatment during the 14-day treatment period. The number of eggs laid, weight of the whole egg and its fractions; daily feed consumption and equivalent drug intake, expressed as the labelled component of Rofenaid; are shown for the SDM and ormetoprim incorporation into the chicken and turkey eggs. The feed consumption and drug intake were corrected for loss in the drinking water pans and spillage, both inside and outside of the layer cages.
Figure 1 shows the typical results of SDM incorporation into the yolk, albumen and shell fractions of the eggs of an 8month old Shamrock White Leghorn chicken pullet. The pullet was fed feed medicated with 0.02% Rofenaid which contained SDM-14C. The average daily intake of labelled drug was equivalent to 9.5 X 107 disintegrations per minute (d.p.m.) per day. The time scale is in days and represents a 14-day treatment period followed by the withdrawal period. All values are expressed as parts per million (p.p.m.) SDM on a logarithmic scale. The concentration of SDM and its possible metabolites plateau at approximately 3.0 p.p.m. in the albumen, 1.5 p.p.m. in the yolk and 1 p.p.m. in the shell fraction. The level of activity in the yolk took longer to build up, and longer to clear, because of the longer time required for yolk development. On the other hand, both the build-up and clearance in both the albumen and shell were extremely rapid, requiring only a few days. The significant level of SDM deposition in the shell was a surprise to us, but may sim-
TABLE 1.—Summary of average biological data for Rofenaid'1' incorporation into chickens and turkeys during the 14-day treatment period Chicken
Turkey
Treatment Compound SDM 3.0 Number of birds Bird weight-kg. 1.67 Number of eggs laid/bird 13.30 Whole egg weight-g. 44.66 Yolk weight-g. 11.41 28.21 Albumen weight-g. Shell weight-g. S.04 101.10 Feed intake/day/bird-g. 13.27 Drug intake/day/bird-mg.
ormetoprim
bSuDi M vi
2.0 1.80 9.50 55.70 17.02 32.86 5.82 99.38 7.83
4.0 11.34 8.75 77.70 22.49 47.43 7.79 206.00 13.35
orm e . " toprim
3.0 11.34 8.00 75.24 23.73 43.60 7.98 180.70 7.03
WITHORAWAL TIME
FIG. 1. SDM deposition into the yolk, albumen and shell fractions of the eggs of an eight-month old Shamrock White Leghorn pullet.
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position and clearance of the ormetoprim portion of Rofenaid in the chicken egg. For the determination of the SDM portion of Rofenaid in the chicken egg, four eightmonth old Shamrock White Leghorn pullets were used. The feed used was Purina Eggena H 16% protein, pelleted layer feed, reground to facilitate the blending of the labelled compound into the feed. For the determination of the deposition and clearance of both the ormetoprim and SDM portions in the turkey egg, the medicated and labelled feed was fed to separate groups of four 11-month old Broad Breasted Bronze turkey hens. A 17% protein turkey layer ration was used.
1183
ROFENAID DEPOSITION AND CLEARANCE
TIME IN DAYS
FIG. 3. Ormetoprim deposition into the yolk, albumen and shell fractions of the eggs of a twoyear old Shaver Leghorn chicken hen.
bumen cleared much more rapidly than the yolk. Figure 4 shows the reproducibility of the uptake and clearance of ormetoprim and its metabolites in the whole egg of two twoyear old Shaver Leghorn chicken hens. Figure 5 shows the typical distribution
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FIG. 2. SDM deposition into the whole eggs of three eight-month old Shamrock White Leghorn pullets.
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Fig. 4. Ormetoprim deposition into the whole eggs of two two-year old Shaver Leghorn chicken hens.
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ply reflect the possibility that SDM can form a calcium salt and may be deposited as such. We did not determine the actual form in which SDM was deposited in the shell. Figure 2 shows the reproducibility of the uptake and clearance of SDM and its metabolites in the whole eggs of three of the four 8-month old Shamrock White Leghorn chicken pullets, as one pullet went out of egg production during the experiment. Figure 3 shows a similar experiment in a two-year old Shaver Leghorn chicken hen, but with the Rofenaid spiked with labelled ormetoprim at a level at which an individual bird would take up approximately 11.1 X 107 d.p.m. per day. The results for ormetoprim incorporation are entirely different from the picture encountered with SDM. The highest levels are found in the yolk at approximately 3.S p.p.m. This undoubtedly reflects the greater lipid solubility of ormetoprim. The albumen levels are over ten times smaller, at approximately 0.25 p.p.m., and the shell levels are approximately 0.07 p.p.m. Again the shell and al-
1184
H. J. LAUEENCOT, A. SCHLOSSER AND J. L. HEMPSTEAD
_•
YOLK X »ALB. — • - o -SHELL
6
10 12 14
6
8
10 12 14
WITHDRAWAL TIME
TIME IN DAYS
FIG. 5. SDM deposition in the yolk, albumen and shell fractions of the eggs of an eleven-month old Broad Breasted Bronze turkey hen.
and concentration of SDM in the eggs of an eleven-month old Broad Breasted Bronze turkey hen. The hen was placed on a medicated feed containing 0.01% Rofenaid labelled with C14 in the SDM portion for fourteen days. The average daily intake was equivalent to 10.1 X 107 d.p.m. per day. The concentration of SDM and its possible metabolites plateau at approximately 1.5 p.p.m. in the albumen, 1.3 p.p.m. in the shell and 0.6 p.p.m. in the yolk. Here, a higher concentration of SDM was found in the shell than in the yolk at the plateau levels. In the chicken the concentration of SDM was higher in the yolk than in the shell at the plateau levels. Again, the levels of activity in the yolk take longer to build up and longer to clear, because of the longer time required for yolk development. The albumen and shell fractions build up and clear rapidly, reflecting their shorter development time. Figure 6 shows the reproducibility of the
• • ' • ' • ' • ' 2 4 6 8 10 12 14 2 TREATMENT TIME
4
I
' 6
8
10 12 14 16 18 20
WITHORAWAL TIME
TIME IN DAYS
Fig. 6. SDM deposition into the whole eggs of four eleven-month old Broad Breasted Bronze turkey hens,
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6
TREATMENT TIME
uptake and clearance of SDM and its metabolites in the whole eggs of four 11month old Broad Breasted turkey hens. Figure 7 shows a similar experiment in an 11-month old Broad Breasted Bronze turkey hen, but with the Rofenaid containing carbon-14 labelled ormetoprim at a level at which an individual bird would take up approximately 63.5 X 106 d.p.m. per day. Here again, as in the chicken, the highest levels of ormetoprim incorporation are found in the yolk at approximately 0.40 p.p.m. The albumen levels are approximately ten times smaller at 0.05 p.p.m., and the shell levels are 0.04 p.p.m. Again, the shell and albumen clear much more rapidly than the yolk. Figure 8 shows the reproducibility of the uptake and clearance of ormetoprim and its metabolites in the whole eggs of two 11month old Broad Breasted Bronze turkey hens. Table 2 summarizes the data for the in-
ROFENAID DEPOSITION AND CLEARANCE
,.
YOLK
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FIG. 8. Ormetoprim deposition into the whole eggs of two eleven-month old Broad Breasted Bronze turkey hens.
concentration of ormetoprim is approximately three times higher than the concentration of SDM. In the yolk of the turkey, the concentration of ormetoprim is less than that of SDM. This lower concentration of ormetoprim found in the turkey egg yolk, as compared to the chicken egg yolk, probably reflects the difference in Rofenaid concentrations in the feed and relative bird size. In the chicken, the feed was medicated at a level of 0.02% Rofenaid. The equivaTABLE 2.—Average incorporation and clearance
of Rofenaid* in chicken and turkey eggs Chicken eggs Parameter
2 4 6 8 10 12 14 2 TREATMENT TIME I
4
6 S 10 12 14 16 18 20 WITHDRAWAL TIME
TIME IN DAYS
Fig. 7. Ormetoprim deposition into the yolk, albumen and shell fractions of the eggs of an eleven-month old Broad Breasted Bronze turkey hen.
ormeSDM toprim
Drug plateau, 7-14 days Whole egg-p.p.m. 1.51 Yolk-p.p.m. 1.20 Albumen-p.p.m. 2.05 Shell-p.p.m. 0.80 Percent of daily uptake at drug plateau in: Whole egg 0.595 Yolk 0.107 0.461 Albumen Shell 0.027 Clearance time to 0.01 p.p m. Whole egg-days 8.6
Turkey eggs SDM
ormetoprim
1.00 3.11 0.25 0.07
1.231 0.612 1.527 1.310
0.166 0.405 0.052 0.042
0.825 0.707 0.114 0.005
0.707 0.103 0.528 0.075
0.174 0.139 0.030 0.005
7.3
13.9
12.1
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corporation of the ormetoprim portion of Rofenaid in the eggs of two Shaver Leghorn chicken hens, three Broad Breasted Bronze turkey hens; and the incorporation of the SDM portion of Rofenaid into three Shamrock White Leghorn chicken pullets and four Broad Breasted Bronze turkey hens. The levels of SDM and ormetoprim are directly compared in the whole eggs and their fractions at the drug plateau, for the percent of the daily drug intake found at the plateau and for the clearance data of the two components of Rofenaid in both the chicken and turkey eggs. The concentration of SDM is higher than that of ormetoprim in the whole eggs of the chickens and turkeys. Among the egg fractions of both the chicken and turkey, the highest concentration of SDM was found in the albumen fraction and the highest concentration of ormetoprim was found in the yolk fraction. In the yolk of the chicken, the
1185
1186
H. J. LAURENCOT, A. SCHLOSSER AND J. L. HEMPSTEAD
DISCUSSION The data from the individual eggs and their fractions reflect the non-uniformity of egg production in regard to development time, total egg weights, or proportional weights of yolk, albumen or shell fractions, and the number of eggs laid in a given sequence or time interval. For these reasons the data from similar fractions of eggs from similarly treated birds cannot be averaged regardless of the similarity of the time of egg laying. The time elements involved in the formation of the yolk, (7-9 days), albumen (25 hours), shell membrane (1 hour) and shell (20 hours) of chicken eggs have been reported with considerably accuracy (Card, 1952). The period of rapid development of the yolk in the turkey has been reported by
Bacon (1970) to require 11 to 14 days. Nestor et al. (1970) have reported a 12 day period of time required for the yolk to develop from a very small size of 0.6 grams to the final size before ovulation. Similar information on turkey eggs albumen and shell development is not available. The time required for the formation of the individual egg fractions is important when studying the clearance of a drug in eggs. When one compares the incorporation, plateau and withdrawal portions of the curves for the albumen and shell fractions for both SDM and ormetoprim incorporation in both the chicken and turkey eggs, we find a rapid uptake, a long plateau level and a rapid clearance in these fractions (Figures 1, 3, 5 and 7). This is due to the relatively short time required for the development of these tissues. On the other hand, the yolk fractions of both labelled components require a longer period of time to reach a plateau level and a longer period of time to clear due to the much longer periods of time required for yolk development. Although the concentration of SDM is lower than that of ormetoprim in the yolk of the chicken and higher in the yolk of the turkey, the longer time required for SDM clearance is more than likely due to the greater absorption and tissue distribution of SDM (Fellig et al., 1971). This greater absorption and higher tissue levels of SDM may contribute for a longer period of time and to a greater extent to the deposition of SDM and its metabolites into a developing yolk, than would be contributed by the lower tissue levels of ormetoprim and its metabolites into a developing yolk. This would account for the observed longer clearance time required for SDM and for the change in the slope of the clearance curves for SDM in the whole egg (Figures 2 and 6) after approximately five days withdrawal from the medicated feed. After five days, the concentration levels shown
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lent intake of ormetoprim per day was 4.35 mg./kg. body weight. In the turkey, the feed was, medicated at a level of 0.01% Rofenaid. The equivalent intake of ormetoprim per day was 0.62 mg./kg. body weight. In the chicken, 0.595% of the daily intake of SDM and 0.825% of the daily intake of ormetoprim were transferred to the whole egg at the plateau level. In the turkey, 0.707% of the daily intake of SDM and 0.174% of the daily intake of ormetoprim were transferred to the whole egg at the plateau level. The distribution of the total activity incorporated into the egg fractions again show the greatest percent incorporation of SDM into the albumen fraction and the greatest percent incorporation of ormetoprim into the yolk fraction. The time required for the whole egg to clear to the 0.01 p.p.m. level was estimated in the chicken at 8.6 days for SDM and 7.3 days for ormetoprim, and in the turkey at 13,9. days for SDM and 12.1 days for ormetoprim.
ROFENAID DEPOSITION AND CLEARANCE
REFERENCES Bacon, W., 1970. The relationship between clutch position and rapid development, yolk weight, and rest in turkey, chicken, and quail egg yolks. Research Summary, 47: 27-28. Ohio Agriculture Research and Development Center, Wooster, Ohio. Card, L. E., 19S2. Poultry Production, Chap. 2. Lea and Febiger, Philadelphia.
Fellig, J., J. Westheimer, M. J. Walsh and W. L. Marusich, 1971. Tissue clearance of Rofenaid in chicken and turkeys. Poultry Sci. SO: 1777-1783. Laurencot, H. J., and J. L. Hempstead, 1971. Liquid scintillation counting of biological materials. I. Solubilized whole blood. In: Organic scintillators and liquid scintillation counting. Edited by Horrocks, D. and C. Peng, p. 635-657. Academic Press, N.Y. Marusich, W. L., E. Ogrinz, M. Brand and M. Mitrovic, 1969. Safety and compatibility of sulfadimethoxine potentiated mixture (Ro 5-0013), a new broad spectrum coccidiostat-antibacterial, in chickens. Poultry Sci. 48: 217-222. Marusich, W. L., E. F. Ogrinz, B. Hecht and M. Mitrovic, 1971. Safety of sulfadimethoxine potentiated mixture (Rofenaid), a new broad spectrum coccidiostat-antibacterial, in turkeys. Poultry Sci. 50: 512-517. Mitrovic, M., E. G. Schildknecht and G. Fusiek, 1969a. Anticoccidial activity of sulfadimethoxine potentiated mixture (Ro 5-0013) in chickens. Poultry Sci. 48: 210-216. Mitrovic, M., G. Fusiek and E. G. Schildknecht, 1969b. Antibacterial activity of sulfadimethoxine potentiated mixture (Ro 5-0013) in chickens. Poultry Sci. 48: 1151-1155. Mitrovic, M., E. G. Schildknecht and G. Fusiek, 1971a. Anticoccidial activity of sulfadimethoxine potentiated mixture (Rofenaid) in turkeys. Poultry Sci. 50: 517-524. Mitrovic, M., G. Fusiek and E. G,.Schildknecht, 1971b. Antibacterial activity of sulfadimethoxine potentiated mixture (Rofenaid) in turkeys. Poultry Sci. 50: 525-529. Nestor, K. E., W. Bacon and P. A. Renner, 1970. Yolk production in egg-type and meat-type turkeys. Research Summary, 47: 29-30. Ohio Agriculture Research and Development Center, Wooster, Ohio.
NEWS AND NOTES (Continued from page 1174) HUBBARD NOTES
AUBURN NOTES
An expansion of hatching facilities for the Hubbard Farms, Little Rock, Arkansas, has been completed. Hatching facilities were increased by 30 percent, and include additional chick sorting space and packing room. The Operation Manager is Wade Helms, and Shannon Skelley is Senior Sales and Service Representative.
The Alabama Poultry Industry Association presented a cheque for $1,748 to Auburn University to sponsor Alabama's 1972 4-H Club ehickenque project conducted by the Cooperative Extension Service. Approximately 1,000 youths participate in the county, district and state levels of competition in the art of barbecuing chicken. A check for $2,500 in support of research on pesti-
(Continued on page 1197)
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represent the activity found in the yolk fraction only, little being contributed by the albumen or shell fraction. These activity levels of SDM and its metabolites found in the yolk at this time reflect not only prior incorporation into the developing yolk from the medicated feed during the very early stages of yolk development, but also incorporation apparently from the relatively high tissue levels of SDM and its metabolites immediately after medicated feed withdrawal, during the early stages of tissue clearance and later stages of the same yolk development. The relatively low tissue plateau concentrations of ormetoprim, which clear faster than the higher tissue plateau concentrations of SDM, apparently do not contribute to as great an extent to the overall activity already incorporated into the developing yolk after five days withdrawal from the medicated feed, as expressed by little change in the slope of the clearance curves for ormetoprim in the whole egg (Figures 4 and 8).
1187
POULTRY SCIENCE 51: 1181-1187, 1972
INTRODUCTION OFENAID®* is a new broad spectrum coccidiostat and antibacterial for poultry. It is a combination of sulfadimethoxine (SDM) (4-sulfanilamido-2,6-dimethoxypyrimidine) and ormetoprim (2,4-diamino-S[ 4,5-dimethoxy- 2 -methylbenzyl ] pyrimidine) in a ratio of 5:3. The anticoccidial activity of Rofenaid has already been reported in the chicken (Mitrovic et al., 1969a) and in the turkey (Mitrovic et al., 1971a), as has also the antibacterial activity in the chicken (Mitrovic et al., 1969b) and in the turkey (Mitrovic et al., 1971b). The safety and compatibility of Rofenaid have been reported in the chicken (Marusich et al., 1969) and in the turkey (Marusich et al., 1971). The present report is concerned with the deposition of Rofenaid into the eggs of laying chicken and
R
* Rofenaid is the Hoffmann-La Roche trademark for a coccidiostat and antibacterial containing sulfadimethoxine and ormetoprim.
turkey hens, and with the length of time required for developing eggs to be clear of the drug, in order to establish a reasonable clearance time for replacement pullets and young breeder hens. MATERIAL AND METHODS Rofenaid, labelled with radioactive carbon-14 either in the SDM or in the ormetoprim portion, was administered to laying chicken hens at a level of 0.02% in the feed, and to laying turkey hens at 0.01% in the feed for 14 days. Eggs were collected daily, separated into yolk, albumen and shell fractions, and assayed for radioactivity by liquid scintillation counting, using a modification of previously reported solubilizing techniques (Laurencot et al., 1971). After the radioactivity had reached a plateau, feeding of the drug was discontinued and analysis of the eggs was continued until no more radioactivity could be detected. Two two-year old Shaver Leghorn hens were used for the determination of the de-
1181
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ABSTRACT Rofenaid® is a coccidiostat and antibacterial for poultry. The active ingredients are a combination of sulfadimethoxine and ormetoprim in a ratio of 5 to 3. The deposition and clearance of Rofenaid® in chicken and turkey eggs were studied in order to establish a clearance time for replacement pullets and young breeder hens. Rofenaid®, labelled with radioactive carbon-" either in the sulfadimethoxine or in the ormetoprim portion was administered to laying chicken hens at a level of 0.02% in the feed, and to laying turkey hens at 0.01% in the feed. Eggs were collected daily, separated into yolk, albumen and shell fractions, and assayed for radioactivity by liquid scintillation counting. When the radioactivity had reached a plateau, feeding of the drug was discontinued and an analysis of the eggs was continued until no more radioactivity could be detected. In the chicken, 0.59% of the daily intake of sulfadimethoxine and 0.83% of the daily intake of ormetoprim were transferred to the whole egg at the plateau level. The time required for the whole egg to clear to the 0.01 p.p.m. level was estimated at 9 days for sulfadimethoxine and 7 days for ormetoprim. In the turkey, 0.71% of the daily intake of sulfadimethoxine and 0.17% of the daily intake of ormetoprim were transferred to the whole egg at the plateau level. The time required for the whole egg to clear to the 0.01 p.p.m. level was estimated at 14 days for sulfadimethoxine and 12 days for ormetoprim.
1182
H. J. LAURENCOT, A. SCHLOSSER AND J. L. HEMPSTEAD
RESULTS In Table 1, the biological data for the four individual treatments are presented. The data are expressed as the average from the individual group of birds used in each treatment during the 14-day treatment period. The number of eggs laid, weight of the whole egg and its fractions; daily feed consumption and equivalent drug intake, expressed as the labelled component of Rofenaid; are shown for the SDM and ormetoprim incorporation into the chicken and turkey eggs. The feed consumption and drug intake were corrected for loss in the drinking water pans and spillage, both inside and outside of the layer cages.
Figure 1 shows the typical results of SDM incorporation into the yolk, albumen and shell fractions of the eggs of an 8month old Shamrock White Leghorn chicken pullet. The pullet was fed feed medicated with 0.02% Rofenaid which contained SDM-14C. The average daily intake of labelled drug was equivalent to 9.5 X 107 disintegrations per minute (d.p.m.) per day. The time scale is in days and represents a 14-day treatment period followed by the withdrawal period. All values are expressed as parts per million (p.p.m.) SDM on a logarithmic scale. The concentration of SDM and its possible metabolites plateau at approximately 3.0 p.p.m. in the albumen, 1.5 p.p.m. in the yolk and 1 p.p.m. in the shell fraction. The level of activity in the yolk took longer to build up, and longer to clear, because of the longer time required for yolk development. On the other hand, both the build-up and clearance in both the albumen and shell were extremely rapid, requiring only a few days. The significant level of SDM deposition in the shell was a surprise to us, but may sim-
TABLE 1.—Summary of average biological data for Rofenaid'1' incorporation into chickens and turkeys during the 14-day treatment period Chicken
Turkey
Treatment Compound SDM 3.0 Number of birds Bird weight-kg. 1.67 Number of eggs laid/bird 13.30 Whole egg weight-g. 44.66 Yolk weight-g. 11.41 28.21 Albumen weight-g. Shell weight-g. S.04 101.10 Feed intake/day/bird-g. 13.27 Drug intake/day/bird-mg.
ormetoprim
bSuDi M vi
2.0 1.80 9.50 55.70 17.02 32.86 5.82 99.38 7.83
4.0 11.34 8.75 77.70 22.49 47.43 7.79 206.00 13.35
orm e . " toprim
3.0 11.34 8.00 75.24 23.73 43.60 7.98 180.70 7.03
WITHORAWAL TIME
FIG. 1. SDM deposition into the yolk, albumen and shell fractions of the eggs of an eight-month old Shamrock White Leghorn pullet.
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position and clearance of the ormetoprim portion of Rofenaid in the chicken egg. For the determination of the SDM portion of Rofenaid in the chicken egg, four eightmonth old Shamrock White Leghorn pullets were used. The feed used was Purina Eggena H 16% protein, pelleted layer feed, reground to facilitate the blending of the labelled compound into the feed. For the determination of the deposition and clearance of both the ormetoprim and SDM portions in the turkey egg, the medicated and labelled feed was fed to separate groups of four 11-month old Broad Breasted Bronze turkey hens. A 17% protein turkey layer ration was used.
1183
ROFENAID DEPOSITION AND CLEARANCE
TIME IN DAYS
FIG. 3. Ormetoprim deposition into the yolk, albumen and shell fractions of the eggs of a twoyear old Shaver Leghorn chicken hen.
bumen cleared much more rapidly than the yolk. Figure 4 shows the reproducibility of the uptake and clearance of ormetoprim and its metabolites in the whole egg of two twoyear old Shaver Leghorn chicken hens. Figure 5 shows the typical distribution
X • Bird 2 0 • Bird 3 a • Bird 4
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FIG. 2. SDM deposition into the whole eggs of three eight-month old Shamrock White Leghorn pullets.
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WITHDRAWAL TIME
TIME IN DAYS
Fig. 4. Ormetoprim deposition into the whole eggs of two two-year old Shaver Leghorn chicken hens.
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ply reflect the possibility that SDM can form a calcium salt and may be deposited as such. We did not determine the actual form in which SDM was deposited in the shell. Figure 2 shows the reproducibility of the uptake and clearance of SDM and its metabolites in the whole eggs of three of the four 8-month old Shamrock White Leghorn chicken pullets, as one pullet went out of egg production during the experiment. Figure 3 shows a similar experiment in a two-year old Shaver Leghorn chicken hen, but with the Rofenaid spiked with labelled ormetoprim at a level at which an individual bird would take up approximately 11.1 X 107 d.p.m. per day. The results for ormetoprim incorporation are entirely different from the picture encountered with SDM. The highest levels are found in the yolk at approximately 3.S p.p.m. This undoubtedly reflects the greater lipid solubility of ormetoprim. The albumen levels are over ten times smaller, at approximately 0.25 p.p.m., and the shell levels are approximately 0.07 p.p.m. Again the shell and al-
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H. J. LAUEENCOT, A. SCHLOSSER AND J. L. HEMPSTEAD
_•
YOLK X »ALB. — • - o -SHELL
6
10 12 14
6
8
10 12 14
WITHDRAWAL TIME
TIME IN DAYS
FIG. 5. SDM deposition in the yolk, albumen and shell fractions of the eggs of an eleven-month old Broad Breasted Bronze turkey hen.
and concentration of SDM in the eggs of an eleven-month old Broad Breasted Bronze turkey hen. The hen was placed on a medicated feed containing 0.01% Rofenaid labelled with C14 in the SDM portion for fourteen days. The average daily intake was equivalent to 10.1 X 107 d.p.m. per day. The concentration of SDM and its possible metabolites plateau at approximately 1.5 p.p.m. in the albumen, 1.3 p.p.m. in the shell and 0.6 p.p.m. in the yolk. Here, a higher concentration of SDM was found in the shell than in the yolk at the plateau levels. In the chicken the concentration of SDM was higher in the yolk than in the shell at the plateau levels. Again, the levels of activity in the yolk take longer to build up and longer to clear, because of the longer time required for yolk development. The albumen and shell fractions build up and clear rapidly, reflecting their shorter development time. Figure 6 shows the reproducibility of the
• • ' • ' • ' • ' 2 4 6 8 10 12 14 2 TREATMENT TIME
4
I
' 6
8
10 12 14 16 18 20
WITHORAWAL TIME
TIME IN DAYS
Fig. 6. SDM deposition into the whole eggs of four eleven-month old Broad Breasted Bronze turkey hens,
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6
TREATMENT TIME
uptake and clearance of SDM and its metabolites in the whole eggs of four 11month old Broad Breasted turkey hens. Figure 7 shows a similar experiment in an 11-month old Broad Breasted Bronze turkey hen, but with the Rofenaid containing carbon-14 labelled ormetoprim at a level at which an individual bird would take up approximately 63.5 X 106 d.p.m. per day. Here again, as in the chicken, the highest levels of ormetoprim incorporation are found in the yolk at approximately 0.40 p.p.m. The albumen levels are approximately ten times smaller at 0.05 p.p.m., and the shell levels are 0.04 p.p.m. Again, the shell and albumen clear much more rapidly than the yolk. Figure 8 shows the reproducibility of the uptake and clearance of ormetoprim and its metabolites in the whole eggs of two 11month old Broad Breasted Bronze turkey hens. Table 2 summarizes the data for the in-
ROFENAID DEPOSITION AND CLEARANCE
,.
YOLK
X • ALB. o • SHELL
< > Bird 5 X • Bird 6
•
2
4
6
8
TREATMENT
'
•
K> 12 TIME TIME
'
<
14
'
2
'
'
'
4
6
a
10
12
WITHDRAWAL
|
14 16 TIME
IN DAYS
FIG. 8. Ormetoprim deposition into the whole eggs of two eleven-month old Broad Breasted Bronze turkey hens.
concentration of ormetoprim is approximately three times higher than the concentration of SDM. In the yolk of the turkey, the concentration of ormetoprim is less than that of SDM. This lower concentration of ormetoprim found in the turkey egg yolk, as compared to the chicken egg yolk, probably reflects the difference in Rofenaid concentrations in the feed and relative bird size. In the chicken, the feed was medicated at a level of 0.02% Rofenaid. The equivaTABLE 2.—Average incorporation and clearance
of Rofenaid* in chicken and turkey eggs Chicken eggs Parameter
2 4 6 8 10 12 14 2 TREATMENT TIME I
4
6 S 10 12 14 16 18 20 WITHDRAWAL TIME
TIME IN DAYS
Fig. 7. Ormetoprim deposition into the yolk, albumen and shell fractions of the eggs of an eleven-month old Broad Breasted Bronze turkey hen.
ormeSDM toprim
Drug plateau, 7-14 days Whole egg-p.p.m. 1.51 Yolk-p.p.m. 1.20 Albumen-p.p.m. 2.05 Shell-p.p.m. 0.80 Percent of daily uptake at drug plateau in: Whole egg 0.595 Yolk 0.107 0.461 Albumen Shell 0.027 Clearance time to 0.01 p.p m. Whole egg-days 8.6
Turkey eggs SDM
ormetoprim
1.00 3.11 0.25 0.07
1.231 0.612 1.527 1.310
0.166 0.405 0.052 0.042
0.825 0.707 0.114 0.005
0.707 0.103 0.528 0.075
0.174 0.139 0.030 0.005
7.3
13.9
12.1
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corporation of the ormetoprim portion of Rofenaid in the eggs of two Shaver Leghorn chicken hens, three Broad Breasted Bronze turkey hens; and the incorporation of the SDM portion of Rofenaid into three Shamrock White Leghorn chicken pullets and four Broad Breasted Bronze turkey hens. The levels of SDM and ormetoprim are directly compared in the whole eggs and their fractions at the drug plateau, for the percent of the daily drug intake found at the plateau and for the clearance data of the two components of Rofenaid in both the chicken and turkey eggs. The concentration of SDM is higher than that of ormetoprim in the whole eggs of the chickens and turkeys. Among the egg fractions of both the chicken and turkey, the highest concentration of SDM was found in the albumen fraction and the highest concentration of ormetoprim was found in the yolk fraction. In the yolk of the chicken, the
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H. J. LAURENCOT, A. SCHLOSSER AND J. L. HEMPSTEAD
DISCUSSION The data from the individual eggs and their fractions reflect the non-uniformity of egg production in regard to development time, total egg weights, or proportional weights of yolk, albumen or shell fractions, and the number of eggs laid in a given sequence or time interval. For these reasons the data from similar fractions of eggs from similarly treated birds cannot be averaged regardless of the similarity of the time of egg laying. The time elements involved in the formation of the yolk, (7-9 days), albumen (25 hours), shell membrane (1 hour) and shell (20 hours) of chicken eggs have been reported with considerably accuracy (Card, 1952). The period of rapid development of the yolk in the turkey has been reported by
Bacon (1970) to require 11 to 14 days. Nestor et al. (1970) have reported a 12 day period of time required for the yolk to develop from a very small size of 0.6 grams to the final size before ovulation. Similar information on turkey eggs albumen and shell development is not available. The time required for the formation of the individual egg fractions is important when studying the clearance of a drug in eggs. When one compares the incorporation, plateau and withdrawal portions of the curves for the albumen and shell fractions for both SDM and ormetoprim incorporation in both the chicken and turkey eggs, we find a rapid uptake, a long plateau level and a rapid clearance in these fractions (Figures 1, 3, 5 and 7). This is due to the relatively short time required for the development of these tissues. On the other hand, the yolk fractions of both labelled components require a longer period of time to reach a plateau level and a longer period of time to clear due to the much longer periods of time required for yolk development. Although the concentration of SDM is lower than that of ormetoprim in the yolk of the chicken and higher in the yolk of the turkey, the longer time required for SDM clearance is more than likely due to the greater absorption and tissue distribution of SDM (Fellig et al., 1971). This greater absorption and higher tissue levels of SDM may contribute for a longer period of time and to a greater extent to the deposition of SDM and its metabolites into a developing yolk, than would be contributed by the lower tissue levels of ormetoprim and its metabolites into a developing yolk. This would account for the observed longer clearance time required for SDM and for the change in the slope of the clearance curves for SDM in the whole egg (Figures 2 and 6) after approximately five days withdrawal from the medicated feed. After five days, the concentration levels shown
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lent intake of ormetoprim per day was 4.35 mg./kg. body weight. In the turkey, the feed was, medicated at a level of 0.01% Rofenaid. The equivalent intake of ormetoprim per day was 0.62 mg./kg. body weight. In the chicken, 0.595% of the daily intake of SDM and 0.825% of the daily intake of ormetoprim were transferred to the whole egg at the plateau level. In the turkey, 0.707% of the daily intake of SDM and 0.174% of the daily intake of ormetoprim were transferred to the whole egg at the plateau level. The distribution of the total activity incorporated into the egg fractions again show the greatest percent incorporation of SDM into the albumen fraction and the greatest percent incorporation of ormetoprim into the yolk fraction. The time required for the whole egg to clear to the 0.01 p.p.m. level was estimated in the chicken at 8.6 days for SDM and 7.3 days for ormetoprim, and in the turkey at 13,9. days for SDM and 12.1 days for ormetoprim.
ROFENAID DEPOSITION AND CLEARANCE
REFERENCES Bacon, W., 1970. The relationship between clutch position and rapid development, yolk weight, and rest in turkey, chicken, and quail egg yolks. Research Summary, 47: 27-28. Ohio Agriculture Research and Development Center, Wooster, Ohio. Card, L. E., 19S2. Poultry Production, Chap. 2. Lea and Febiger, Philadelphia.
Fellig, J., J. Westheimer, M. J. Walsh and W. L. Marusich, 1971. Tissue clearance of Rofenaid in chicken and turkeys. Poultry Sci. SO: 1777-1783. Laurencot, H. J., and J. L. Hempstead, 1971. Liquid scintillation counting of biological materials. I. Solubilized whole blood. In: Organic scintillators and liquid scintillation counting. Edited by Horrocks, D. and C. Peng, p. 635-657. Academic Press, N.Y. Marusich, W. L., E. Ogrinz, M. Brand and M. Mitrovic, 1969. Safety and compatibility of sulfadimethoxine potentiated mixture (Ro 5-0013), a new broad spectrum coccidiostat-antibacterial, in chickens. Poultry Sci. 48: 217-222. Marusich, W. L., E. F. Ogrinz, B. Hecht and M. Mitrovic, 1971. Safety of sulfadimethoxine potentiated mixture (Rofenaid), a new broad spectrum coccidiostat-antibacterial, in turkeys. Poultry Sci. 50: 512-517. Mitrovic, M., E. G. Schildknecht and G. Fusiek, 1969a. Anticoccidial activity of sulfadimethoxine potentiated mixture (Ro 5-0013) in chickens. Poultry Sci. 48: 210-216. Mitrovic, M., G. Fusiek and E. G. Schildknecht, 1969b. Antibacterial activity of sulfadimethoxine potentiated mixture (Ro 5-0013) in chickens. Poultry Sci. 48: 1151-1155. Mitrovic, M., E. G. Schildknecht and G. Fusiek, 1971a. Anticoccidial activity of sulfadimethoxine potentiated mixture (Rofenaid) in turkeys. Poultry Sci. 50: 517-524. Mitrovic, M., G. Fusiek and E. G,.Schildknecht, 1971b. Antibacterial activity of sulfadimethoxine potentiated mixture (Rofenaid) in turkeys. Poultry Sci. 50: 525-529. Nestor, K. E., W. Bacon and P. A. Renner, 1970. Yolk production in egg-type and meat-type turkeys. Research Summary, 47: 29-30. Ohio Agriculture Research and Development Center, Wooster, Ohio.
NEWS AND NOTES (Continued from page 1174) HUBBARD NOTES
AUBURN NOTES
An expansion of hatching facilities for the Hubbard Farms, Little Rock, Arkansas, has been completed. Hatching facilities were increased by 30 percent, and include additional chick sorting space and packing room. The Operation Manager is Wade Helms, and Shannon Skelley is Senior Sales and Service Representative.
The Alabama Poultry Industry Association presented a cheque for $1,748 to Auburn University to sponsor Alabama's 1972 4-H Club ehickenque project conducted by the Cooperative Extension Service. Approximately 1,000 youths participate in the county, district and state levels of competition in the art of barbecuing chicken. A check for $2,500 in support of research on pesti-
(Continued on page 1197)
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represent the activity found in the yolk fraction only, little being contributed by the albumen or shell fraction. These activity levels of SDM and its metabolites found in the yolk at this time reflect not only prior incorporation into the developing yolk from the medicated feed during the very early stages of yolk development, but also incorporation apparently from the relatively high tissue levels of SDM and its metabolites immediately after medicated feed withdrawal, during the early stages of tissue clearance and later stages of the same yolk development. The relatively low tissue plateau concentrations of ormetoprim, which clear faster than the higher tissue plateau concentrations of SDM, apparently do not contribute to as great an extent to the overall activity already incorporated into the developing yolk after five days withdrawal from the medicated feed, as expressed by little change in the slope of the clearance curves for ormetoprim in the whole egg (Figures 4 and 8).
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