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Incubation of Avian Eggs in an Inverted Position
RESEARCH NOTES
1223
TABLE 1.—Effect of thyroidectomy on the hydroxyproline content of the femoral vein in the rooster
Deep femoral v Femur Proximal caudal femoral v
Surgery
Wef weight
Water content
Dry fat-free weight
,i , "ydroxy"rol,ne % of dry weight
embrrjnosus m
ddle caudal femoral iafic n afic a
Distal caudal femoral \ Popliteal v
Internal femoral v
Sartorlus m
Fic. 1. Dissection of the femoral vein in the rooster, Gallus domesticus.
Time after surgery (6 weeks vs. 9 weeks) was not a factor in altering vein composition. The data for both time periods were therefore pooled. Neither collagen nor water content was significantly altered by thyroidectomy. In the chicken, thyroxine and thyrotrophin do not seem to be major factors in the regulation of collagen and water content of veins.
Control left vein right vein
104.0+ 8' 108.6+11
Thyroidectomized left vein 100.5 + 10 right vein 100.4+ 8
50.5+0.4 61.1+0.7
41.8+4 42.3+4
12.1+0.1 12.0+0.2
60.6+1 61.2 + 1
42.0+4 42.4+4
12.02+0.1 12.15+0.1
1 Each value represents the mean ± standard error of the mean of observations on 6 animals.
REFERENCES Asboe-Hansen, G., 1959. Endocrine control of connective tissue. Am. J. Med. 26: 4/0-484. Hisaw, F. L., and M. X. Zarrow, 1950. The physiology of relaxin. Vit. Horm. 8: 151-178. Neumann, R. E., and M. A. Logan, 1950a. The determination of hydroxyproline. J. Biol. Chem. 184: 299 306. Neumann, R. E., and M. A. Logan, 1950b. The determination of collagen and elastin in tissues. J. Biol. Chem. 186: 549-556. Svejcar, J., J. Prerovsky, J. Linhart and J. Kruml, 1963. Content of collagen, elastin, and hexosamine in primary varicose veins. Clin. Sci. 24: 325-330. Zarrow, M. X., and J. Yochim, 1961. Dilation of the uterine cervix of the rat accompanying changes during the estrous cycle, pregnancy, and following treatment with estrogen, progesterone, and relaxin. End. 69 : 292-304.
INCUBATION OF AVIAN EGGS IN AN INVERTED POSITION J. R. CAIN AND U. K. ABBOTT Department of Avian Sciences, University of California, Davis, California 95616 (Received for publication M a r c h 16, 1971)
INTRODUCTION
Egg position during incubation has been considered an important factor in satisfactory hatchability and one which influences the incidence of certain embryo malpositions (Landauer, 1967). In particular, incubation of eggs in an inverted position, i.e., with the small end up (S.E.U.) has
been reported to drastically reduce hatchability and to result in about a 60% incidence of embryos in Malposition 2 (head in small end of the egg) and 5 (feet over head) (Byerly and Olsen, 1931, 1933). Reports in the literature indicate hatchabilities as low as 36.4% (Byerly and Olsen, 1933) although recently El-Ibiary et al.
1224
RESEARCH NOTES
(1966) reported a much less drastic reduction in hatch, using mostly eggs from S.C. White Leghorn pullets. Homma and Shumiya (1964), however, reported that in Japanese quail hatchability was sharply reduced in eggs incubated in an inverted position. We have compared the effects of incubating in the SEU position eggs from three avian species, coturnix quail, turkeys and chickens from several very different sources. MATERIALS AND METHOD
Coturnix (Japanese quail) eggs came from U.C.D. line 908 (Sittmann et al, 1966). The turkey eggs came from a commercial strain of Large Whites obtained from Nicholas Turkey Breeding Farms. The chicken eggs were from commercial strains and from experimental lines. A White Leghorn line in the fourth generation of selection for a high incidence of double-yolked (D.Y.) was chosen because of its variability in egg size and shape. In this case pedigree data available permitted an analysis of hatchability by sister families. The scaleless stock (Abbott and Asmundson, 1962) represented another source of material not selected for egg characteristics, exhibiting considerable variation in this respect and of particular interest because a substantial proportion of the birds lay rather long narrow eggs. The commercial White Leghorn eggs included those from a line obtained a number of years ago from Kimber Farms and two groups obtained directly from DeKalb, one from older hens and a second from pullets. All eggs were incubated in commercial Jamesway 252 incubators at standard conditions of temperature and humidity for each species. Special coturnix inserts were used to adapt chicken egg size trays for the smaller coturnix eggs (Wilson et al., 1961). Eggs were either in the normal (large end up) or the inverted (small end up) position
until transfer time; turning through 90° occurred every 2 hours. During the last 3 days eggs from both treatment groups were transferred to hatching baskets and were in a horizontal position. Eggs were candled twice during incubation; all infertiles and dead embryos were broken out and examined. Late dead-inshell embryos and those that pipped but failed to hatch were scored as: in a normal position for their stage of development; in Malposition 2; Malposition 5; or in one of the other malpositions (Sanctuary, 1924; Insko, 1949). RESULTS AND DISCUSSION
Table 1 indicates clearly that the commercial strains of chickens used were least affected by incubation in the S.E.U. position. Thus hatchability in both DeKalb hens and pullets was reduced about 16% but was still nearly 80%. Very similar results were obtained with the eggs of Kimber stock. Both of the U.C. experimental lines hatched at a considerably lower rate than the high performing commercial lines and both showed a greater reduction in hatchability when incubated in the S.E.U. position. The reduction in hatch was especially marked in the D.Y. line. The analysis by sister families within this line indicated that the reduction in hatchability ranged from 6% to 62% and that hatchability obtained in the normal position was not related in any consistent fashion with that in the inverted position. The commercial turkey line and most especially our coturnix were drastically affected by incubation in the inverted position. Hatchability of turkey eggs was reduced to 33%, a decrease of 40% from control figures, while Japanese quail incubated in the S.E.U. position hatched only about ^ as well as their controls. The majority of late dead-in-shell embryos and those in which pipping had
1225
RESEARCH NOTES TABLE 1.—Effects Species & Line Coturnix 908 Turkey Lg. White Chickens U. C. D.Y. Chickens Scaleless Chickens Kimber Chickens DeKalb Chickens DeKalb Pullets a
dead 0 to 3 days;
of incubation in a small-end-up position (S.E.U.) on hatchability
Treatment
No. Fert.
/o Hatchab.
Dg. 1"
Dg. 21=
%
% DIS«
Pips
Control S.E.U. Control S.E.U. Control S.E.U. Control S.E.U. Control S.E.U. Control S.E.U. Control S.E.U.
639 617 178 263 957 1029 64 334 99 318 178 789 178 411
73.1 17.0 73.6 33.1 76.8 47.0 64.1 44.9 90.9 78.9 94.9 78.8 93.8 78.7
7.0 7.4 10.1 9.9 7.9 11.1 21.8 22.2 3.0 3.2 1.7 2.8 0 4.2
3.9 5.2 2.3 6.1 2.9 4.7 3.2 2.7 1.0 0.6 0.6 1.2 2.8 2.0
7.8 24.8 9.5 31.2 7.1 24.7 6.2 13.5 2.0 6.6 1.7 6.7 2.8 4.9
8.1 45.5 4.5 19.8 4.3 13.0 4.7 16.8 3.0 10.7 1.1 10.5 0.6 10.2
b
dead 4 days to transfer;
e
barely commenced were in Malposition 2; this was especially noticeable in coturnix. Those embryos able to proceed progressively further in pipping were found in Malposition S or in a normal position. This seems to suggest that at transfer time, the embryos had their heads in the small end of the egg, but that many of these were able to assume a quasi-normal orientation during the hatching process. Many embryos which remained with head in the small end were also successful in emerging from the shell. In conclusion, these results indicate that the three avian species tested vary markedly in their response to incubation in the inverted position, with Japanese quail eggs being most affected, turkey eggs intermediate and chickens most resistant. The data also indicate that different chicken lines may respond very differently to this treatment. Thus, the least reduction in hatchability was obtained with eggs from highproducing commercial sources and the greatest decrease from the two U.C. experimental lines in which egg shape and hatchability have not been under any selection pressure. That considerable between-family variation in response can exist was shown by the analysis conducted with the double
%
%
dead in shell.
yolk line, which also indicated that hatchability in the normal position per se was not positively correlated with degree of reduction in the S.E.U. position. It seems evident that the selection and crossing procedures used to develop the high hatching commercial strains have also resulted in embryos better able to hatch in sub-optimal environments. It is likely that the more uniform egg shapes achieved in such stocks play an important role in this regard. In marked contrast, turkey eggs in which selection for hatchability and egg characteristics has been largely neglected in favor of growth and conformation, proved to be much less resistant to this incubation stress. Similarly, coturnix have not been directly selected for hatchability or egg characteristics and here again their poor performance also appears to emphasize the importance of these traits. Perhaps this comparative study can in part reconcile the rather different results reported in the older literature. Clearly the results of such a study depend on the characteristics of the stock used. REFERENCES Abbott, U. K., and V. S. Asmundson, 1962. Responses to selection under severe environmental
1226
RESEARCH NOTES
stress. Proc. Xllth World's Poultry Congr., Sydney, Australia: 30. Byerly, T. C , and M. W. Olsen, 1931. The influence of gravity and air-hunger on hatchability. Poultry Sci. 10: 281-287. Byerly, T. C , and M. W. Olsen, 1933. Time and manner of determination of the malposition head-in-small-end-of-egg. Poultry Sci. 12: 261265. El-Ibiary, H. M., C. S. Shaffner and E. F. Godfrey, 1966. Hatchability of eggs set small end up. Poultry Sci. 45: 419-420. Homma, K., and S. Shumiya, 1964. Studies on the hatchability of the Japanese quail; 1. Orientation of the egg and frequencies of malpositions. Jap. J. Animal Reprod. 10: 81-84. Landauer, W., 1967. The hatchability of chicken
eggs as influenced by environment and heredity. Monograph 1 (Rev.). Univ. Conn., Agr. Exp. Sta., Storrs, Conn. Sanctuary, W. C , 1924. One cause of dead chicks in the shell. Poultry Sci. 4 : 141-143. Sittmann, K., H. Abplanalp and R. A. Fraser, 1966. Inbreeding depression in Japanese quail. Genetics, 54: 371-379. Insko, W. M., Jr., 1949. Physical conditions in incubation. In: L. W. Taylor., (ed.) Fertility and hatchability of chicken and turkey eggs. John Wiley and Sons, Inc., N.Y., p. 317. Wilson, W. O., U. K. Abbott and H. Abplanalp, 1961. Evaluation of coturnix (Japanese quail) as pilot animal for poultry. Poultry Sci. 40: 651-657.
COMPARISON OF ORGANOCHLORINE RESIDUE LEVELS IN CHICKEN PIECES, GIBLETS, AND BRAIN1 MARY E. ZABIK AND KAYE FUNK Department of Food Science and Human Nutrition, Michigan State University, East Lansing, Michigan 48823 (Received for publication March 25, 1971) INTRODUCTION
Cummings et al. (1967) reported low level feeding of selected pesticides to hens resulted in levels up to 10 times the level fed in adipose tissue while liver accumulated levels approximately equal to those fed and breast muscle had levels of ^ of the amount in the feed. While the present investigators were studying the effect of cooking on thigh meat and skins from hens fed feed contaminated with 25 p.p.m. of lindane, dieldrin and DDT, it was noted that pesticide concentrations based on fat content were approximately equal in both raw tissues and about twice the values in the feed. It was decided to compare pesticide levels in the raw meat and skin with 1
Michigan Agricultural Journal No. 5420.
Experiment
Station
those of the raw giblets and brain to compare distribution following high levels of pesticide feeding. EXPERIMENTAL Ten White Leghorn hens, approximately 10 months of age, were fed ad libitum a diet contaminated with 25 p.p.m. each of analytical grade lindane, dieldrin, and p,p'DDT for 5 weeks prior to slaughter. Thigh meat, thigh skin, heart, liver, gizzards, and brain tissue were analyzed for pesticide residues. Using hexane-acetone extraction, acetonitrile partitioning and Florisil-Celite column clean-up procedures described by Funk et al. (1971) and electron capture GLC conditions as outlined by Zabik and Dugan (1971) residue levels were expressed as p.p.m. in the fat of each tissue.
1223
TABLE 1.—Effect of thyroidectomy on the hydroxyproline content of the femoral vein in the rooster
Deep femoral v Femur Proximal caudal femoral v
Surgery
Wef weight
Water content
Dry fat-free weight
,i , "ydroxy"rol,ne % of dry weight
embrrjnosus m
ddle caudal femoral iafic n afic a
Distal caudal femoral \ Popliteal v
Internal femoral v
Sartorlus m
Fic. 1. Dissection of the femoral vein in the rooster, Gallus domesticus.
Time after surgery (6 weeks vs. 9 weeks) was not a factor in altering vein composition. The data for both time periods were therefore pooled. Neither collagen nor water content was significantly altered by thyroidectomy. In the chicken, thyroxine and thyrotrophin do not seem to be major factors in the regulation of collagen and water content of veins.
Control left vein right vein
104.0+ 8' 108.6+11
Thyroidectomized left vein 100.5 + 10 right vein 100.4+ 8
50.5+0.4 61.1+0.7
41.8+4 42.3+4
12.1+0.1 12.0+0.2
60.6+1 61.2 + 1
42.0+4 42.4+4
12.02+0.1 12.15+0.1
1 Each value represents the mean ± standard error of the mean of observations on 6 animals.
REFERENCES Asboe-Hansen, G., 1959. Endocrine control of connective tissue. Am. J. Med. 26: 4/0-484. Hisaw, F. L., and M. X. Zarrow, 1950. The physiology of relaxin. Vit. Horm. 8: 151-178. Neumann, R. E., and M. A. Logan, 1950a. The determination of hydroxyproline. J. Biol. Chem. 184: 299 306. Neumann, R. E., and M. A. Logan, 1950b. The determination of collagen and elastin in tissues. J. Biol. Chem. 186: 549-556. Svejcar, J., J. Prerovsky, J. Linhart and J. Kruml, 1963. Content of collagen, elastin, and hexosamine in primary varicose veins. Clin. Sci. 24: 325-330. Zarrow, M. X., and J. Yochim, 1961. Dilation of the uterine cervix of the rat accompanying changes during the estrous cycle, pregnancy, and following treatment with estrogen, progesterone, and relaxin. End. 69 : 292-304.
INCUBATION OF AVIAN EGGS IN AN INVERTED POSITION J. R. CAIN AND U. K. ABBOTT Department of Avian Sciences, University of California, Davis, California 95616 (Received for publication M a r c h 16, 1971)
INTRODUCTION
Egg position during incubation has been considered an important factor in satisfactory hatchability and one which influences the incidence of certain embryo malpositions (Landauer, 1967). In particular, incubation of eggs in an inverted position, i.e., with the small end up (S.E.U.) has
been reported to drastically reduce hatchability and to result in about a 60% incidence of embryos in Malposition 2 (head in small end of the egg) and 5 (feet over head) (Byerly and Olsen, 1931, 1933). Reports in the literature indicate hatchabilities as low as 36.4% (Byerly and Olsen, 1933) although recently El-Ibiary et al.
1224
RESEARCH NOTES
(1966) reported a much less drastic reduction in hatch, using mostly eggs from S.C. White Leghorn pullets. Homma and Shumiya (1964), however, reported that in Japanese quail hatchability was sharply reduced in eggs incubated in an inverted position. We have compared the effects of incubating in the SEU position eggs from three avian species, coturnix quail, turkeys and chickens from several very different sources. MATERIALS AND METHOD
Coturnix (Japanese quail) eggs came from U.C.D. line 908 (Sittmann et al, 1966). The turkey eggs came from a commercial strain of Large Whites obtained from Nicholas Turkey Breeding Farms. The chicken eggs were from commercial strains and from experimental lines. A White Leghorn line in the fourth generation of selection for a high incidence of double-yolked (D.Y.) was chosen because of its variability in egg size and shape. In this case pedigree data available permitted an analysis of hatchability by sister families. The scaleless stock (Abbott and Asmundson, 1962) represented another source of material not selected for egg characteristics, exhibiting considerable variation in this respect and of particular interest because a substantial proportion of the birds lay rather long narrow eggs. The commercial White Leghorn eggs included those from a line obtained a number of years ago from Kimber Farms and two groups obtained directly from DeKalb, one from older hens and a second from pullets. All eggs were incubated in commercial Jamesway 252 incubators at standard conditions of temperature and humidity for each species. Special coturnix inserts were used to adapt chicken egg size trays for the smaller coturnix eggs (Wilson et al., 1961). Eggs were either in the normal (large end up) or the inverted (small end up) position
until transfer time; turning through 90° occurred every 2 hours. During the last 3 days eggs from both treatment groups were transferred to hatching baskets and were in a horizontal position. Eggs were candled twice during incubation; all infertiles and dead embryos were broken out and examined. Late dead-inshell embryos and those that pipped but failed to hatch were scored as: in a normal position for their stage of development; in Malposition 2; Malposition 5; or in one of the other malpositions (Sanctuary, 1924; Insko, 1949). RESULTS AND DISCUSSION
Table 1 indicates clearly that the commercial strains of chickens used were least affected by incubation in the S.E.U. position. Thus hatchability in both DeKalb hens and pullets was reduced about 16% but was still nearly 80%. Very similar results were obtained with the eggs of Kimber stock. Both of the U.C. experimental lines hatched at a considerably lower rate than the high performing commercial lines and both showed a greater reduction in hatchability when incubated in the S.E.U. position. The reduction in hatch was especially marked in the D.Y. line. The analysis by sister families within this line indicated that the reduction in hatchability ranged from 6% to 62% and that hatchability obtained in the normal position was not related in any consistent fashion with that in the inverted position. The commercial turkey line and most especially our coturnix were drastically affected by incubation in the inverted position. Hatchability of turkey eggs was reduced to 33%, a decrease of 40% from control figures, while Japanese quail incubated in the S.E.U. position hatched only about ^ as well as their controls. The majority of late dead-in-shell embryos and those in which pipping had
1225
RESEARCH NOTES TABLE 1.—Effects Species & Line Coturnix 908 Turkey Lg. White Chickens U. C. D.Y. Chickens Scaleless Chickens Kimber Chickens DeKalb Chickens DeKalb Pullets a
dead 0 to 3 days;
of incubation in a small-end-up position (S.E.U.) on hatchability
Treatment
No. Fert.
/o Hatchab.
Dg. 1"
Dg. 21=
%
% DIS«
Pips
Control S.E.U. Control S.E.U. Control S.E.U. Control S.E.U. Control S.E.U. Control S.E.U. Control S.E.U.
639 617 178 263 957 1029 64 334 99 318 178 789 178 411
73.1 17.0 73.6 33.1 76.8 47.0 64.1 44.9 90.9 78.9 94.9 78.8 93.8 78.7
7.0 7.4 10.1 9.9 7.9 11.1 21.8 22.2 3.0 3.2 1.7 2.8 0 4.2
3.9 5.2 2.3 6.1 2.9 4.7 3.2 2.7 1.0 0.6 0.6 1.2 2.8 2.0
7.8 24.8 9.5 31.2 7.1 24.7 6.2 13.5 2.0 6.6 1.7 6.7 2.8 4.9
8.1 45.5 4.5 19.8 4.3 13.0 4.7 16.8 3.0 10.7 1.1 10.5 0.6 10.2
b
dead 4 days to transfer;
e
barely commenced were in Malposition 2; this was especially noticeable in coturnix. Those embryos able to proceed progressively further in pipping were found in Malposition S or in a normal position. This seems to suggest that at transfer time, the embryos had their heads in the small end of the egg, but that many of these were able to assume a quasi-normal orientation during the hatching process. Many embryos which remained with head in the small end were also successful in emerging from the shell. In conclusion, these results indicate that the three avian species tested vary markedly in their response to incubation in the inverted position, with Japanese quail eggs being most affected, turkey eggs intermediate and chickens most resistant. The data also indicate that different chicken lines may respond very differently to this treatment. Thus, the least reduction in hatchability was obtained with eggs from highproducing commercial sources and the greatest decrease from the two U.C. experimental lines in which egg shape and hatchability have not been under any selection pressure. That considerable between-family variation in response can exist was shown by the analysis conducted with the double
%
%
dead in shell.
yolk line, which also indicated that hatchability in the normal position per se was not positively correlated with degree of reduction in the S.E.U. position. It seems evident that the selection and crossing procedures used to develop the high hatching commercial strains have also resulted in embryos better able to hatch in sub-optimal environments. It is likely that the more uniform egg shapes achieved in such stocks play an important role in this regard. In marked contrast, turkey eggs in which selection for hatchability and egg characteristics has been largely neglected in favor of growth and conformation, proved to be much less resistant to this incubation stress. Similarly, coturnix have not been directly selected for hatchability or egg characteristics and here again their poor performance also appears to emphasize the importance of these traits. Perhaps this comparative study can in part reconcile the rather different results reported in the older literature. Clearly the results of such a study depend on the characteristics of the stock used. REFERENCES Abbott, U. K., and V. S. Asmundson, 1962. Responses to selection under severe environmental
1226
RESEARCH NOTES
stress. Proc. Xllth World's Poultry Congr., Sydney, Australia: 30. Byerly, T. C , and M. W. Olsen, 1931. The influence of gravity and air-hunger on hatchability. Poultry Sci. 10: 281-287. Byerly, T. C , and M. W. Olsen, 1933. Time and manner of determination of the malposition head-in-small-end-of-egg. Poultry Sci. 12: 261265. El-Ibiary, H. M., C. S. Shaffner and E. F. Godfrey, 1966. Hatchability of eggs set small end up. Poultry Sci. 45: 419-420. Homma, K., and S. Shumiya, 1964. Studies on the hatchability of the Japanese quail; 1. Orientation of the egg and frequencies of malpositions. Jap. J. Animal Reprod. 10: 81-84. Landauer, W., 1967. The hatchability of chicken
eggs as influenced by environment and heredity. Monograph 1 (Rev.). Univ. Conn., Agr. Exp. Sta., Storrs, Conn. Sanctuary, W. C , 1924. One cause of dead chicks in the shell. Poultry Sci. 4 : 141-143. Sittmann, K., H. Abplanalp and R. A. Fraser, 1966. Inbreeding depression in Japanese quail. Genetics, 54: 371-379. Insko, W. M., Jr., 1949. Physical conditions in incubation. In: L. W. Taylor., (ed.) Fertility and hatchability of chicken and turkey eggs. John Wiley and Sons, Inc., N.Y., p. 317. Wilson, W. O., U. K. Abbott and H. Abplanalp, 1961. Evaluation of coturnix (Japanese quail) as pilot animal for poultry. Poultry Sci. 40: 651-657.
COMPARISON OF ORGANOCHLORINE RESIDUE LEVELS IN CHICKEN PIECES, GIBLETS, AND BRAIN1 MARY E. ZABIK AND KAYE FUNK Department of Food Science and Human Nutrition, Michigan State University, East Lansing, Michigan 48823 (Received for publication March 25, 1971) INTRODUCTION
Cummings et al. (1967) reported low level feeding of selected pesticides to hens resulted in levels up to 10 times the level fed in adipose tissue while liver accumulated levels approximately equal to those fed and breast muscle had levels of ^ of the amount in the feed. While the present investigators were studying the effect of cooking on thigh meat and skins from hens fed feed contaminated with 25 p.p.m. of lindane, dieldrin and DDT, it was noted that pesticide concentrations based on fat content were approximately equal in both raw tissues and about twice the values in the feed. It was decided to compare pesticide levels in the raw meat and skin with 1
Michigan Agricultural Journal No. 5420.
Experiment
Station
those of the raw giblets and brain to compare distribution following high levels of pesticide feeding. EXPERIMENTAL Ten White Leghorn hens, approximately 10 months of age, were fed ad libitum a diet contaminated with 25 p.p.m. each of analytical grade lindane, dieldrin, and p,p'DDT for 5 weeks prior to slaughter. Thigh meat, thigh skin, heart, liver, gizzards, and brain tissue were analyzed for pesticide residues. Using hexane-acetone extraction, acetonitrile partitioning and Florisil-Celite column clean-up procedures described by Funk et al. (1971) and electron capture GLC conditions as outlined by Zabik and Dugan (1971) residue levels were expressed as p.p.m. in the fat of each tissue.