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Determination of the Age of Bruises in Chicken Broilers1
Determination of the Age of Bruises in Chicken Broilers1 M. K. HAMDY, K. N. MAY, W. P. FLANAGAN AND J. J. POWERS Food Technology & Poultry Departments, University of Georgia, Athens, Georgia (Received for publication August 10, 1960)
1 Journal Paper Number 134, College Experiment Station, University of Georgia, College of Agriculture Experiment Stations, Athens, Georgia. •Fouche's reagent is made by mixing 25 g. of
an earlier publication (Hamdy et al., 1957). Morphological Changes: The progression of gross and pathological changes occurring in livestock bruises were reported by Hamdy et al. (1957) and Rickenbacker (1959). Hamdy et al. (1957) stated that the time required for these gross changes to develop was variable but that their sequence was fairly consistent. Morphological studies on poultry bruises confirmed these previous observations except for the time element which was found to be very short compared with human, rabbit, or livestock bruises, apparently due to the high metabolic rate and other physiological factors (body temperature, heart rate). The sequence of the visible changes of poultry bruises was found to be subject to variation due to changes in environmental temperatures as noted in Table 1. Spectrophotometric examination of extracts of poultry bruises (Hamdy et al., 1960) showed that hemoglobin was degraded mostly to biliverdin in birds kept at 86°F. and mostly to bilirubin in birds kept at 70°F. or below. These bile pigments were isolated and biliverdin was found to be responsible for the green color whereas bilirubin formed the yellow-orange color in the bruised tissue. Determination of the Age of a Bruise: The principle of this method is based on previous findings (Hamdy et al., 1960) that bilirubin or biliverdin is formed during the healing process of poultry bruises as a result of the catabolism of hemoglobin present in the damaged areas and that trichloricoacetic acid, 10 ml. of 10% ferric chloride, and 100 ml. of distilled water (Naumann, 1936).
787
Downloaded from http://ps.oxfordjournals.org/ at New York University on April 16, 2015
' I VHE loss to the commercial broiler in-*- dustry due to bruising accounts for many millions of dollars annually. If the age of these bruises could be easily ascertained, a means would then be available for assigning the responsibility for the losses involved and possibly developing incentives for minimizing such losses. At present, the determination of the age of poultry bruises is based on gross observations of the characteristic progression of color alterations in the damaged area. These color changes have been shown to vary considerably due to many physical and physiological factors (Hamdy et al., 1960). It has been reported (Hamdy et al., 1957) that a chemical test based on the presence of bilirubin, a bile pigment resulting from the hemoglobin degradation in bruised tissues, can be used to estimate the age of livestock bruises. The present work was carried out to examine the possible use of this color test for the determination of the age of poultry bruises and use this method to determine the percentage distribution according to the age of bruises trimmed from broilers in a federally inspected processing plant. Bruising and Sampling: Methods of inflicting standard type bruises and for procuring tissue samples for examination were previously reported (Hamdy et al., 1960). Detection of Bilirubin: The bruised and control tissues were excised immediately after slaughtering the birds, scored, and immersed in Fouche* reagent as outlined in
788
M. K. HAMDY, K. N. MAY, W. P. FLANAGAN AND J. J. POWERS TABLE 1.—The effect of environmental temperatures on the external and internal gross appearance of poultry bruises as related to lime*
Age of bruise** Control
Temperature*** (°F.)
External Color
Swelling
—
70 86
Fluid Color
Volume
Normal Normal
—
Normal Normal
12 hrs.
70 86
24 hrs.
70 86
36 hrs.
70 86
-
Yel low-green-purple Diffused, green-purple
++ ++ +++ +++ +++ ++ + +
48 hrs.
70 86
-
Yellow-green (orange) Dark-green
+ +
Orange Dark-green yellow
72 hrs.
70 86
-
Yellow-orange Almost normal
+ +
Yellow Slight yellow
96 hrs.
70 86
—
Slight yellow Normal
—
120 hrs.
70 86
-
Normal Normal
—
2 min.
70 86
Red Red Diffused dark red-purple Diffused red-purple Diffused light green-purple Diffused light green-purple
Red Red Dark red Dark-red-purple Brown Brown-slight yellow Yellow-orange Yellow-slight yellow
— —
* Results based on 780 birds. ** All the birds were bruised using the same force. *** Temperature of chicken house.
bilirubin can be detected in the tissues by Fouche's reagent (Hamdy et al., 19S7) in which bilirubin is oxidized to biliverdin by the ferric chloride in the presence of the trichloroacetic acid solution (Naumann, 1936). The results of this method, applied to experimentally inflicted poultry bruises
as shown in Table 2, indicate the absence of chemically detectable bile pigment in control (non-bruised) tissues and in bruises less than 13 hours old. The color reaction of the bruised tissues was diffused light blue after 13-14 hours, diffused light green at 24-36 hours, and dark green at 2-3 days
TABLE 2.—The color of poultry bruises, after reaction with FoucM reag as related to the age of the bruise* Age of bruise
Tissue color after 20 minutes in Fouche's reagent
Normal tissue (control) Zero-13 hrs. 14-24 hrs. 24-36 hrs.
No color. Pink within minutes turning brown. Diffused light blue along with pink and brown. Diffused light green especially.at the periphery with a brown color (in the center) of the bruise. Diffused dark green along with brown center. Dark green spots or crystals imbedded in the bruised area. No brown. No evidence of damaged area. Sometimes slight blue color could be detected.
3 days 4 days 5 days
* The bruises were inflicted on the breast, using a standard technique.
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++ ++ +++ +++ + +
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AGE OF BRUISES TABLE 3.—Percentage distribution of 1,812 condemned bruises according to age
TABLE 4.—Percentage distribution of 1,402 condemned bruises according to anatomical location
Age of bruise
Percent of total
Location of bruise
Percent of total
0-12 hrs. 12-24 hrs. 24-36 hrs. Over 2 days
90 7.5 2.0 0.5
Breast Wing Thigh Leg Leg & thigh
38 32 3 17 10
tors in a commercial processing plant and the results (Table 3 and 4) indicate that 90% of the bruises were inflicted between zero and 13 hours before slaughtering and that 38% of the examined tissues were breast bruises. ACKNOWLEDGMENT
Bruised tissue samples for part of this work were supplied by Athens Poultry Inc., Athens, Georgia. REFERENCES Hamdy, M. K., L. W. Kunkle and F. E. Deatherage, 1957. Bruised tissue II. Determination of the age of a bruise. J. Animal Sci. 16: 490-495. Hamdy, M. K., J. J. Powers and K. N. May, 1960. Some biochemical changes occurring in experimentally inflicted bruises. Food Technol. 14 (4): 46. Naumann, H. N., 1936. CIX. Studies on bile pigments II. A new test for bilirubin in the urine and its use of detection of bilirubin in normal urine. Biochem. J. 30: 762-764. Rickenbacker, J. E., 1959. Biochemical problems in determining the age of bruised animal tissue. Farmer Cooperative Service, U. S. Department of Agriculture. Service Report 42.
NEWS AND NOTES (Continued from page 771) Members of the Committee of Fifty elected were: V. Pringle, Harrisonburg, Virginia; and T. B. Avery, Manhattan, Kansas, to succeed T. Heitz and J. L. Perry. XIITH WORLD'S POULTRY CONGRESS International Travel Service, Inc., 119 South
State Street, Chicago, Illinois, has been appointed by the United States and Canadian Committees to operate tours and coordinate travel arrangements for Congress participation from both countries in connection with the XHth World's Poultry Congress to be held in Sydney, Australia, August 13-18, 1962.
(Continued on page 807)
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after contusion. Dark green crystals imbeded in the damaged areas were observed 3-4 days after contusion probably due to high local concentration caused by the phagocytic activities of the macrophages. Alkaline hematin produced a pink color turning brown whereas hemoglobin produced dark brown color in the Fouche reagent. The diffused light blue color was found to be due to a concentration of 50100 ppm. bilirubin albuminate adsorbed onto the tissues (Hamdy et al., 1957) and the color was intensified to a dark green when the tissue adsorbed high concentration of bilirubin. The sensitivity and specificity of this chemical test were found to be the same as those reported in an earlier publication (Hamdy et al., 1957). For best results it is recommended that this test be applied immediately after slaughtering the birds by trimming the bruised area with one inch of normal tissue surrounding the bruise and scoring it to facilitate the penetration of the Fouche reagent. This test was applied on condemned parts of poultry carcasses which had been excised by the U.S.D.A. inspec-
1 Journal Paper Number 134, College Experiment Station, University of Georgia, College of Agriculture Experiment Stations, Athens, Georgia. •Fouche's reagent is made by mixing 25 g. of
an earlier publication (Hamdy et al., 1957). Morphological Changes: The progression of gross and pathological changes occurring in livestock bruises were reported by Hamdy et al. (1957) and Rickenbacker (1959). Hamdy et al. (1957) stated that the time required for these gross changes to develop was variable but that their sequence was fairly consistent. Morphological studies on poultry bruises confirmed these previous observations except for the time element which was found to be very short compared with human, rabbit, or livestock bruises, apparently due to the high metabolic rate and other physiological factors (body temperature, heart rate). The sequence of the visible changes of poultry bruises was found to be subject to variation due to changes in environmental temperatures as noted in Table 1. Spectrophotometric examination of extracts of poultry bruises (Hamdy et al., 1960) showed that hemoglobin was degraded mostly to biliverdin in birds kept at 86°F. and mostly to bilirubin in birds kept at 70°F. or below. These bile pigments were isolated and biliverdin was found to be responsible for the green color whereas bilirubin formed the yellow-orange color in the bruised tissue. Determination of the Age of a Bruise: The principle of this method is based on previous findings (Hamdy et al., 1960) that bilirubin or biliverdin is formed during the healing process of poultry bruises as a result of the catabolism of hemoglobin present in the damaged areas and that trichloricoacetic acid, 10 ml. of 10% ferric chloride, and 100 ml. of distilled water (Naumann, 1936).
787
Downloaded from http://ps.oxfordjournals.org/ at New York University on April 16, 2015
' I VHE loss to the commercial broiler in-*- dustry due to bruising accounts for many millions of dollars annually. If the age of these bruises could be easily ascertained, a means would then be available for assigning the responsibility for the losses involved and possibly developing incentives for minimizing such losses. At present, the determination of the age of poultry bruises is based on gross observations of the characteristic progression of color alterations in the damaged area. These color changes have been shown to vary considerably due to many physical and physiological factors (Hamdy et al., 1960). It has been reported (Hamdy et al., 1957) that a chemical test based on the presence of bilirubin, a bile pigment resulting from the hemoglobin degradation in bruised tissues, can be used to estimate the age of livestock bruises. The present work was carried out to examine the possible use of this color test for the determination of the age of poultry bruises and use this method to determine the percentage distribution according to the age of bruises trimmed from broilers in a federally inspected processing plant. Bruising and Sampling: Methods of inflicting standard type bruises and for procuring tissue samples for examination were previously reported (Hamdy et al., 1960). Detection of Bilirubin: The bruised and control tissues were excised immediately after slaughtering the birds, scored, and immersed in Fouche* reagent as outlined in
788
M. K. HAMDY, K. N. MAY, W. P. FLANAGAN AND J. J. POWERS TABLE 1.—The effect of environmental temperatures on the external and internal gross appearance of poultry bruises as related to lime*
Age of bruise** Control
Temperature*** (°F.)
External Color
Swelling
—
70 86
Fluid Color
Volume
Normal Normal
—
Normal Normal
12 hrs.
70 86
24 hrs.
70 86
36 hrs.
70 86
-
Yel low-green-purple Diffused, green-purple
++ ++ +++ +++ +++ ++ + +
48 hrs.
70 86
-
Yellow-green (orange) Dark-green
+ +
Orange Dark-green yellow
72 hrs.
70 86
-
Yellow-orange Almost normal
+ +
Yellow Slight yellow
96 hrs.
70 86
—
Slight yellow Normal
—
120 hrs.
70 86
-
Normal Normal
—
2 min.
70 86
Red Red Diffused dark red-purple Diffused red-purple Diffused light green-purple Diffused light green-purple
Red Red Dark red Dark-red-purple Brown Brown-slight yellow Yellow-orange Yellow-slight yellow
— —
* Results based on 780 birds. ** All the birds were bruised using the same force. *** Temperature of chicken house.
bilirubin can be detected in the tissues by Fouche's reagent (Hamdy et al., 19S7) in which bilirubin is oxidized to biliverdin by the ferric chloride in the presence of the trichloroacetic acid solution (Naumann, 1936). The results of this method, applied to experimentally inflicted poultry bruises
as shown in Table 2, indicate the absence of chemically detectable bile pigment in control (non-bruised) tissues and in bruises less than 13 hours old. The color reaction of the bruised tissues was diffused light blue after 13-14 hours, diffused light green at 24-36 hours, and dark green at 2-3 days
TABLE 2.—The color of poultry bruises, after reaction with FoucM reag as related to the age of the bruise* Age of bruise
Tissue color after 20 minutes in Fouche's reagent
Normal tissue (control) Zero-13 hrs. 14-24 hrs. 24-36 hrs.
No color. Pink within minutes turning brown. Diffused light blue along with pink and brown. Diffused light green especially.at the periphery with a brown color (in the center) of the bruise. Diffused dark green along with brown center. Dark green spots or crystals imbedded in the bruised area. No brown. No evidence of damaged area. Sometimes slight blue color could be detected.
3 days 4 days 5 days
* The bruises were inflicted on the breast, using a standard technique.
Downloaded from http://ps.oxfordjournals.org/ at New York University on April 16, 2015
++ ++ +++ +++ + +
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AGE OF BRUISES TABLE 3.—Percentage distribution of 1,812 condemned bruises according to age
TABLE 4.—Percentage distribution of 1,402 condemned bruises according to anatomical location
Age of bruise
Percent of total
Location of bruise
Percent of total
0-12 hrs. 12-24 hrs. 24-36 hrs. Over 2 days
90 7.5 2.0 0.5
Breast Wing Thigh Leg Leg & thigh
38 32 3 17 10
tors in a commercial processing plant and the results (Table 3 and 4) indicate that 90% of the bruises were inflicted between zero and 13 hours before slaughtering and that 38% of the examined tissues were breast bruises. ACKNOWLEDGMENT
Bruised tissue samples for part of this work were supplied by Athens Poultry Inc., Athens, Georgia. REFERENCES Hamdy, M. K., L. W. Kunkle and F. E. Deatherage, 1957. Bruised tissue II. Determination of the age of a bruise. J. Animal Sci. 16: 490-495. Hamdy, M. K., J. J. Powers and K. N. May, 1960. Some biochemical changes occurring in experimentally inflicted bruises. Food Technol. 14 (4): 46. Naumann, H. N., 1936. CIX. Studies on bile pigments II. A new test for bilirubin in the urine and its use of detection of bilirubin in normal urine. Biochem. J. 30: 762-764. Rickenbacker, J. E., 1959. Biochemical problems in determining the age of bruised animal tissue. Farmer Cooperative Service, U. S. Department of Agriculture. Service Report 42.
NEWS AND NOTES (Continued from page 771) Members of the Committee of Fifty elected were: V. Pringle, Harrisonburg, Virginia; and T. B. Avery, Manhattan, Kansas, to succeed T. Heitz and J. L. Perry. XIITH WORLD'S POULTRY CONGRESS International Travel Service, Inc., 119 South
State Street, Chicago, Illinois, has been appointed by the United States and Canadian Committees to operate tours and coordinate travel arrangements for Congress participation from both countries in connection with the XHth World's Poultry Congress to be held in Sydney, Australia, August 13-18, 1962.
(Continued on page 807)
Downloaded from http://ps.oxfordjournals.org/ at New York University on April 16, 2015
after contusion. Dark green crystals imbeded in the damaged areas were observed 3-4 days after contusion probably due to high local concentration caused by the phagocytic activities of the macrophages. Alkaline hematin produced a pink color turning brown whereas hemoglobin produced dark brown color in the Fouche reagent. The diffused light blue color was found to be due to a concentration of 50100 ppm. bilirubin albuminate adsorbed onto the tissues (Hamdy et al., 1957) and the color was intensified to a dark green when the tissue adsorbed high concentration of bilirubin. The sensitivity and specificity of this chemical test were found to be the same as those reported in an earlier publication (Hamdy et al., 1957). For best results it is recommended that this test be applied immediately after slaughtering the birds by trimming the bruised area with one inch of normal tissue surrounding the bruise and scoring it to facilitate the penetration of the Fouche reagent. This test was applied on condemned parts of poultry carcasses which had been excised by the U.S.D.A. inspec-