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The Effect of Carbon Dioxide Inhalation and Sodium Bicarbonate Ingestion on Egg Shell Deposition
Research Notes THE EFFECT OF CARBON DIOXIDE INHALATION AND SODIUM BICARBONATE INGESTION ON EGG SHELL DEPOSITION F. R. FRANK AND R. E. BURGER Department of Animal Physiology, and Department of Poultry University of California, Davis
Husbandry,
(Received for publication July 2, 1965)
This work was supported in part by Training Grant GM-646 from the Division of General Medical Sciences, U.S.P.H.S.
ventilating fan and controlled humidity. Atmospheric carbon dioxide tensions were increased at regular intervals from 0.3 mm. Hg to 19 mm. Hg over a 21-day period and then maintained at 19 mm. Hg for 21 days. Arterial blood samples were drawn by cardiac puncture at the end of the six-week period in the first trial. Egg weights and numbers, shell thickness, shell weights, and water consumption were measured in both trials. Egg production was maintained at high levels, 74 percent before and 71.3 percent during chronic exposure to carbon dioxide inhalation. Figure 1 shows shell thickness was a function of the level of carbon dioxide tension maintained in the chamber, increasing some 12 percent over the normal at 19 mm. Hg. The observed increase in egg weight of 1.04 gm. was not significant (P < 0.2). Time of oviposition was not altered. An uncompensated respiratory acidosis was observed; pH, 7.48 and 7.40; bicarbonate ion concentration, 20.8 and 21.2 mM/ 1; arterial carbon dioxide tension, 28.3 and 34.9 mm. Hg, respectively, for 0.3 mm. Hg and 19 mm. Hg carbon dioxide tension of the inspired air. Sodium bicarbonate ingestion, as with carbon dioxide inhalation, was predicted to lead to hypercapnia due to respiratory compensation for the induced metabolic alkalosis but to be accompanied by either normal or slightly higher blood pH values. Sodium bicarbonate was added to the
1604
Downloaded from http://ps.oxfordjournals.org/ at New York University on April 22, 2015
Helbacka et al. (1963) reported that exposure of laying hens to an atmosphere containing S percent carbon dioxide for 24 to 48 hours led to a marked decrease in the calcification of the shell. They suggested that deposition of calcium carbonate is dependent upon blood pH and is depressed when blood pH is lowered either by respiratory acidosis or by ammonium chloride feeding (Hall and Helbacka, 1959). This suggestion that depressed deposition of calcite follows low blood pH, independent of arterial carbon dioxide tension, is difficult to reconcile with results obtained with mammalian species. Freeman and Fenn (19S3) found that rats chronically exposed to high levels of inspired carbon dioxide displayed hypercalcifkation of the bone, calcite being the major mineral component deposited in excess over the normal. Schaefer et al. (1963) exposed subjects chronically to 1.5 percent carbon dioxide and suggested that the disturbance noted in calcium metabolism was dependent upon a similar hypermineralization of the bone. In the present study, ten laying hens, 50 weeks old, in each of two experiments were exposed to atmospheres containing regulated levels of carbon dioxide in a 5' by 5' by 12' chamber equipped with individual cages, automated lights and waterers, a
160S
RESEARCH NOTES Sh. Th. -3Z8+.W%0 r~. 76 * ' .10
$heii thickness, mm.
20
Carbon dioxide tension t /^
/
mm. Jiff.
FIG. 1. The dependency of shell thickness on the level of environmental carbon dioxide tension. In the regression equation, shell thickness (Sh.Th.) is in units of 0.01 mm. and carbon dioxide tension of the environment (Pco ) is in mm. Hg.
drinking water in an attempt to prevent the expected diuresis from flushing the added sodium ion out of the blood stream. Levels of sodium bicarbonate were increased in drinking water from 0.2 S percent to 1.00 percent over a four-week period and maintained at 1.00 percent for three weeks. At this time arterial blood samples were drawn, and pH and carbon dioxide tensions measured. A diuresis was observed, increasing in intensity as the level of sodium bicarbonate was increased. Egg production, egg weight, and shell thickness did not significantly alter. Blood pH, carbon dioxide, and bicarbonate ion concentration in arterial blood were not different from the control values indicating that when hens were given access to normal levels of chloride in the diet, extra sodium ion given in the drinking water was not retained. Another experiment was designed to limit the intake of chloride ion so that the concentration of bicarbonate ion in blood must increase to maintain total anion equi-
valence to sodium concentrations. A low sodium chloride diet (0.08 percent Na) was fed to a new sample of twelve 50-week-old hens. Initially, sodium chloride was given to the hens in the drinking water at a level of 0.25 percent. After 2 weeks on this regime and when normal egg production (75 percent) and egg shell thickness (0.315 mm.) were observed, sodium bicarbonate at 0.25 percent was substituted for sodium chloride in the drinking water. A diuresis was observed that lasted for about four days. After this period, presumably when excess chloride ion was excreted, the diuresis diminished, and a normal intake of water, as sodium bicarbonate solution, was observed. After two weeks, shell thickness was increased by 6.6 percent. Further increases in sodium bicarbonate levels were not accompanied by further increases in shell thickness. A marked diuresis was not observed even when the level of sodium bicarbonate reached 1 percent in the drinking water. These results tend to substantiate the
Downloaded from http://ps.oxfordjournals.org/ at New York University on April 22, 2015
.&
1606
RESEARCH NOTES ACKNOWLEDGMENT
The authors wish to express their gratitude to D. Lumijarvi for his technical assistance. REFERENCES Freeman, F. H., and W. O. Fenn, 1953. Changes in carbon dioxide stores of rats due to atmospheres low in oxygen or high in carbon dioxide. Am. J. Physiol. 174: 422-430. Hall, K. N., and N. V. Helbacka, 1959. Improving albumen quality. Poultry Sci. 38: 111-114. Helbacka, N. V., J. L. Casterline and C. J. Smith, 1963. The effect of high C0 2 atmospheres on the laying hen. Poultry Sci. 42: 1082-1084. Hunt, J. R., and J. R. Aitken, 1962. The effect of ammonium and chloride ions in the diet of hens on egg shell quality. Poultry Sci. 4 1 : 434438. Schaefer, K. E., B. J. Hastings, C. R. Carey and G. Nichols, Jr., 1963. Respiratory acclimatization to carbon dioxide. J. Appl. Physiol. 18: 1079-1084.
CLINICAL SALMONELLOSIS ACCIDENTALLY INDUCED BY FEED AND WATER DEPRIVATION OF ONE-WEEK-OLD BROILER CHICKS B. W. BIERER AND T. H. ELEAZER The South Carolina Agricultural Experiment Station, Clemson University, Clemson, S.C. (Received for publication July 3, 1965)
Bierer (1960) found that mortality in poults artificially exposed to Salmonella typhimurium infection was greatest when exposure occurred during the first day, was less severe with exposure on the second and third days, and was nonexistent if exposure occurred after the third day. Bierer (1961a) reported that chicks artificially exposed to S. pullorum experienced a higher mortality when brooder temperature was 26.7°C compared to chicks similarly exposed but brooded at 32.2°C. Deaths in 100 pen mate chicks not inoculated were 40% in the low temperature group, cornPublished with permission of the director as Technical Contribution No. 544.
pared to 1% in the chicks brooded at 32.2°C. Bierer (1961b) was unable to satisfactorily induce clinical 5. typhimurium infection by per os inoculation of baby chicks, but succeeded when the bacteria were sprayed on pipping eggs in the incubator. Williams (1959) listed water starvation as one of a number of items sometimes associated with paratyphoid infection under field conditions. In this report the ill effects of feed and water deprivation were being evaluated when postmortem lesions of Salmonellosis were observed in certain groups undertreatment. Twenty battery brooders, each containing 10 one-week-old chicks, were arranged
Downloaded from http://ps.oxfordjournals.org/ at New York University on April 22, 2015
hypothesis of Hunt and Aitken (1962) that the reduction of shell thickness following ammonium chloride-induced metabolic acidosis is through the limiting of blood bicarbonate ion for calcite deposition. However, we feel these effects on shell deposition are not mediated through constituents of the blood directly; the effects are probably mediated through alterations of the bicarbonate buffer within the lumen of the shell gland which is isolated from blood by at least two cells and their membranes. In summary, chronic exposures to regimes designed to increase blood carbon dioxide levels and/or blood bicarbonate levels lead to increases in shell thickness regardless of effects on blood pH. Therefore, blood pH per se cannot be implicated in the regulation of calcification of the egg shell.
Husbandry,
(Received for publication July 2, 1965)
This work was supported in part by Training Grant GM-646 from the Division of General Medical Sciences, U.S.P.H.S.
ventilating fan and controlled humidity. Atmospheric carbon dioxide tensions were increased at regular intervals from 0.3 mm. Hg to 19 mm. Hg over a 21-day period and then maintained at 19 mm. Hg for 21 days. Arterial blood samples were drawn by cardiac puncture at the end of the six-week period in the first trial. Egg weights and numbers, shell thickness, shell weights, and water consumption were measured in both trials. Egg production was maintained at high levels, 74 percent before and 71.3 percent during chronic exposure to carbon dioxide inhalation. Figure 1 shows shell thickness was a function of the level of carbon dioxide tension maintained in the chamber, increasing some 12 percent over the normal at 19 mm. Hg. The observed increase in egg weight of 1.04 gm. was not significant (P < 0.2). Time of oviposition was not altered. An uncompensated respiratory acidosis was observed; pH, 7.48 and 7.40; bicarbonate ion concentration, 20.8 and 21.2 mM/ 1; arterial carbon dioxide tension, 28.3 and 34.9 mm. Hg, respectively, for 0.3 mm. Hg and 19 mm. Hg carbon dioxide tension of the inspired air. Sodium bicarbonate ingestion, as with carbon dioxide inhalation, was predicted to lead to hypercapnia due to respiratory compensation for the induced metabolic alkalosis but to be accompanied by either normal or slightly higher blood pH values. Sodium bicarbonate was added to the
1604
Downloaded from http://ps.oxfordjournals.org/ at New York University on April 22, 2015
Helbacka et al. (1963) reported that exposure of laying hens to an atmosphere containing S percent carbon dioxide for 24 to 48 hours led to a marked decrease in the calcification of the shell. They suggested that deposition of calcium carbonate is dependent upon blood pH and is depressed when blood pH is lowered either by respiratory acidosis or by ammonium chloride feeding (Hall and Helbacka, 1959). This suggestion that depressed deposition of calcite follows low blood pH, independent of arterial carbon dioxide tension, is difficult to reconcile with results obtained with mammalian species. Freeman and Fenn (19S3) found that rats chronically exposed to high levels of inspired carbon dioxide displayed hypercalcifkation of the bone, calcite being the major mineral component deposited in excess over the normal. Schaefer et al. (1963) exposed subjects chronically to 1.5 percent carbon dioxide and suggested that the disturbance noted in calcium metabolism was dependent upon a similar hypermineralization of the bone. In the present study, ten laying hens, 50 weeks old, in each of two experiments were exposed to atmospheres containing regulated levels of carbon dioxide in a 5' by 5' by 12' chamber equipped with individual cages, automated lights and waterers, a
160S
RESEARCH NOTES Sh. Th. -3Z8+.W%0 r~. 76 * ' .10
$heii thickness, mm.
20
Carbon dioxide tension t /^
/
mm. Jiff.
FIG. 1. The dependency of shell thickness on the level of environmental carbon dioxide tension. In the regression equation, shell thickness (Sh.Th.) is in units of 0.01 mm. and carbon dioxide tension of the environment (Pco ) is in mm. Hg.
drinking water in an attempt to prevent the expected diuresis from flushing the added sodium ion out of the blood stream. Levels of sodium bicarbonate were increased in drinking water from 0.2 S percent to 1.00 percent over a four-week period and maintained at 1.00 percent for three weeks. At this time arterial blood samples were drawn, and pH and carbon dioxide tensions measured. A diuresis was observed, increasing in intensity as the level of sodium bicarbonate was increased. Egg production, egg weight, and shell thickness did not significantly alter. Blood pH, carbon dioxide, and bicarbonate ion concentration in arterial blood were not different from the control values indicating that when hens were given access to normal levels of chloride in the diet, extra sodium ion given in the drinking water was not retained. Another experiment was designed to limit the intake of chloride ion so that the concentration of bicarbonate ion in blood must increase to maintain total anion equi-
valence to sodium concentrations. A low sodium chloride diet (0.08 percent Na) was fed to a new sample of twelve 50-week-old hens. Initially, sodium chloride was given to the hens in the drinking water at a level of 0.25 percent. After 2 weeks on this regime and when normal egg production (75 percent) and egg shell thickness (0.315 mm.) were observed, sodium bicarbonate at 0.25 percent was substituted for sodium chloride in the drinking water. A diuresis was observed that lasted for about four days. After this period, presumably when excess chloride ion was excreted, the diuresis diminished, and a normal intake of water, as sodium bicarbonate solution, was observed. After two weeks, shell thickness was increased by 6.6 percent. Further increases in sodium bicarbonate levels were not accompanied by further increases in shell thickness. A marked diuresis was not observed even when the level of sodium bicarbonate reached 1 percent in the drinking water. These results tend to substantiate the
Downloaded from http://ps.oxfordjournals.org/ at New York University on April 22, 2015
.&
1606
RESEARCH NOTES ACKNOWLEDGMENT
The authors wish to express their gratitude to D. Lumijarvi for his technical assistance. REFERENCES Freeman, F. H., and W. O. Fenn, 1953. Changes in carbon dioxide stores of rats due to atmospheres low in oxygen or high in carbon dioxide. Am. J. Physiol. 174: 422-430. Hall, K. N., and N. V. Helbacka, 1959. Improving albumen quality. Poultry Sci. 38: 111-114. Helbacka, N. V., J. L. Casterline and C. J. Smith, 1963. The effect of high C0 2 atmospheres on the laying hen. Poultry Sci. 42: 1082-1084. Hunt, J. R., and J. R. Aitken, 1962. The effect of ammonium and chloride ions in the diet of hens on egg shell quality. Poultry Sci. 4 1 : 434438. Schaefer, K. E., B. J. Hastings, C. R. Carey and G. Nichols, Jr., 1963. Respiratory acclimatization to carbon dioxide. J. Appl. Physiol. 18: 1079-1084.
CLINICAL SALMONELLOSIS ACCIDENTALLY INDUCED BY FEED AND WATER DEPRIVATION OF ONE-WEEK-OLD BROILER CHICKS B. W. BIERER AND T. H. ELEAZER The South Carolina Agricultural Experiment Station, Clemson University, Clemson, S.C. (Received for publication July 3, 1965)
Bierer (1960) found that mortality in poults artificially exposed to Salmonella typhimurium infection was greatest when exposure occurred during the first day, was less severe with exposure on the second and third days, and was nonexistent if exposure occurred after the third day. Bierer (1961a) reported that chicks artificially exposed to S. pullorum experienced a higher mortality when brooder temperature was 26.7°C compared to chicks similarly exposed but brooded at 32.2°C. Deaths in 100 pen mate chicks not inoculated were 40% in the low temperature group, cornPublished with permission of the director as Technical Contribution No. 544.
pared to 1% in the chicks brooded at 32.2°C. Bierer (1961b) was unable to satisfactorily induce clinical 5. typhimurium infection by per os inoculation of baby chicks, but succeeded when the bacteria were sprayed on pipping eggs in the incubator. Williams (1959) listed water starvation as one of a number of items sometimes associated with paratyphoid infection under field conditions. In this report the ill effects of feed and water deprivation were being evaluated when postmortem lesions of Salmonellosis were observed in certain groups undertreatment. Twenty battery brooders, each containing 10 one-week-old chicks, were arranged
Downloaded from http://ps.oxfordjournals.org/ at New York University on April 22, 2015
hypothesis of Hunt and Aitken (1962) that the reduction of shell thickness following ammonium chloride-induced metabolic acidosis is through the limiting of blood bicarbonate ion for calcite deposition. However, we feel these effects on shell deposition are not mediated through constituents of the blood directly; the effects are probably mediated through alterations of the bicarbonate buffer within the lumen of the shell gland which is isolated from blood by at least two cells and their membranes. In summary, chronic exposures to regimes designed to increase blood carbon dioxide levels and/or blood bicarbonate levels lead to increases in shell thickness regardless of effects on blood pH. Therefore, blood pH per se cannot be implicated in the regulation of calcification of the egg shell.