ENVIRONMENT AND HEALTH Altered Metabolism of Carotenoids During Pale-Bird Syndrome in Chickens Infected with Eimeria acervulina1 JULIUSZ K. TYCZKOWSKI and PAT B. HAMILTON2 Department of Poultry Science, North Carolina State University, Raleigh, North Carolina 27695-7608 MICHAEL D. RUFF Animal Parasitology Institute, USDA, Agricultural Research Service, Beltsville Agricultural Research Center, West, Beltsville, Maryland 20705 (Received for publication November 8, 1990)
1991 Poultry Science 70:2074-2081 INTRODUCTION
Pale-bird syndrome (PBS) is a synonym for generalized malabsorption syndrome, but in its strictest sense it refers to abnormal pigmentation. The yellow color of poultry carcasses and egg yolks is provided by oxycarotenoids that are not synthesized de novo by poultry and must be absorbed from the botanical components of their diets (Marusich and Bauernfeind, 1981). The utilization of dietary oxycarotenoids by poultry is affected by many agents such as aflatoxin (Tyczkowski and Hamilton, 1987), ochratoxin (Schaeffer et a/., 1987), reoviruses (Ruff and Rosenberger, 1985), and coccidia (Ruff, 1986) or interactions between coccidia and mycotoxins (Huff and Ruff, 1982) or coccidia and reoviruses (Ruff and
The use of trade names in this publication does not imply endorsement by the North Carolina Agricultural Research Service of the products mentioned, nor criticism of similar ones not mentioned. ^"o whom correspondence should be addressed.
Rosenberger, 1985). The coccidial species Eimeria acervulina causes a severe depression of serum oxycarotenoids (Ruff and Fuller, 1975), which correlates widi a decrease in integumentary oxycarotenoids and with a decrease in perceived color (Dua et al., 1967). This consequence of E. acervulina infection is related presumably to the fact that E. acervulina infections are localized in the duodenum and upper jejunum (Ruff and Fuller, 1975), which are the sites for absorption of oxycarotenoids in chickens (Tyczkowski and Hamilton, 1986a). The prospect of studying PBS caused by coccidiosis in more detail than previously possible was offered by the development of HPLC methods for the analysis of different forms and isomers of carotenoids in diets and tissues (Tyczkowski and Hamilton, 1984) and by the development of experimental models in which known oxycarotenoids are fed instead of the customary unknown mixtures (Tyczkowski and Hamilton, 1986b). The objective of the present study was to investigate the progression of changes in tissue carotenoids of
2074
Downloaded from http://ps.oxfordjournals.org/ at Oakland University on June 3, 2015
ABSTRACT The progression of changes in carotenoid metabolism during pale-bird syndrome caused by a coccidial infection was investigated. Male broiler chickens 15 days of age on a yellow corn and soybean meal-based diet were infected with Eimeria acervulina oocysts and their serum, liver, and toe webs were sampled at 0,4, 6, and 10 days postinfection for HPLC analysis of carotenoids. At 4 days postinfection a drastic reduction (71%) in serum lutein, the main body carotenoid, and smaller reductions in liver (58%) and toe webs (38%) occurred. Derivative forms of lutein, mainly esters, continued to be lost from tissues for 10 days postinfection. These carotenoids were apparently lost via the intestinal tract because birds placed on a white corn and soybean meal-based diet at time of infection had lutein in their jejunal contents even at 7 days postinfection. The loss of carotenoids from the body was accompanied by a decreasing ability to absorb canthaxanthin, a red carotenoid, from the intestinal contents. The absorption of canthaxanthin measured at 0, 3, 4, 5, 6, and 7 days reached its low point of 1% of preinfection ability on Day 5 before a slow recovery commenced. Thus, the pale-bird syndrome caused by E. acervulina appeared to be the result of a loss of previously absorbed carotenoids coupled with drastic malabsorption of dietary carotenoids. (Key words: Eimeria acervulina, lutein, malabsorption, carotenoids, excretion)
CAROTENOID METABOLISM AND EIMERIA ACERVUUNA
chickens during infection with E. acervulina in order to determine whether the PBS caused by E. acervulina is the result of an efflux of oxycarotenoids from the serum and tissues, an inability to absorb oxycarotenoids from the intestinal tract, or a combination of both mechanisms. MATERIALS AND METHODS
Husbandry
Survey of Tissue Carotenoids During Infection At 0, 4, 6, and 10 days postinfection, four groups of four birds were bled (2.5 mL per bird) for serum collection, killed by cervical dislocation, and their livers and one foot were removed. The samples of each tissue were combined on a group basis and stored at -20 C prior to analysis. The toe webs of the feet after thawing were sampled with a circular (3 mm diameter) punch (Marusich and Bauernfeind, 1981) to provide four circles per foot. The livers were homogenized in deionized water (1:5, wt/vol).
3
Roxanthin Red 10, Hoffinann-LaRoche, Inc., Nulley, NJ 07110.
TABLE 1. Composition of diet Ingredients and calculated analysis Ground com1 Soybean meal (48.5% CP) Cottonseed oil Dicalcium phosphate (21% Ca, 18% P) Ground limestone Salt DL-methionine (98%) Vitamin premix Mineral premix Calculated analysis: Protein, % Fat, % ME, kcal per kg
Percentage 62.22 32.00 2.00 2.50 .60 .32 .26 .05 .05 21.00 4.50 3,041
1 The corn was yellow or white as specified for the experiment 2 Vitamin premix provided the following per kilogram of diet: vitamin A, 6,600 IU; cholecalciferol, 3,300 IU; vitamin E, 5.5 IU; niacin, 44 mg; pantothenic acid, 11 mg; riboflavin, 6.6 mg; menadione sodium bisulfite, 4.4 mg; pyridoxine, 1.1 mg; thiamine, 1.1 mg; folic acid, .22 mg; biotin, 28 Hg; vitamin Bj2, 13 Mg; selenium, .10 mg. 3 Mineral premix provided the following per kilogram of diet: Mn, 60 mg; Zn, 60 mg; Fe, 20 mg; Cu, 4.4 mg; I, .35 mg; Co, .05 mg.
Egress and Absorption of Carotenoids During Infection In another experiment, birds at 0, 3,4, and 5 days postinfection were removed from the yellow com and soybean meal-based diet. They were placed on a white corn and soybean mealbased diet (Table 1) essentially free of lutein (
1991 Poultry Science 70:2074-2081 INTRODUCTION
Pale-bird syndrome (PBS) is a synonym for generalized malabsorption syndrome, but in its strictest sense it refers to abnormal pigmentation. The yellow color of poultry carcasses and egg yolks is provided by oxycarotenoids that are not synthesized de novo by poultry and must be absorbed from the botanical components of their diets (Marusich and Bauernfeind, 1981). The utilization of dietary oxycarotenoids by poultry is affected by many agents such as aflatoxin (Tyczkowski and Hamilton, 1987), ochratoxin (Schaeffer et a/., 1987), reoviruses (Ruff and Rosenberger, 1985), and coccidia (Ruff, 1986) or interactions between coccidia and mycotoxins (Huff and Ruff, 1982) or coccidia and reoviruses (Ruff and
The use of trade names in this publication does not imply endorsement by the North Carolina Agricultural Research Service of the products mentioned, nor criticism of similar ones not mentioned. ^"o whom correspondence should be addressed.
Rosenberger, 1985). The coccidial species Eimeria acervulina causes a severe depression of serum oxycarotenoids (Ruff and Fuller, 1975), which correlates widi a decrease in integumentary oxycarotenoids and with a decrease in perceived color (Dua et al., 1967). This consequence of E. acervulina infection is related presumably to the fact that E. acervulina infections are localized in the duodenum and upper jejunum (Ruff and Fuller, 1975), which are the sites for absorption of oxycarotenoids in chickens (Tyczkowski and Hamilton, 1986a). The prospect of studying PBS caused by coccidiosis in more detail than previously possible was offered by the development of HPLC methods for the analysis of different forms and isomers of carotenoids in diets and tissues (Tyczkowski and Hamilton, 1984) and by the development of experimental models in which known oxycarotenoids are fed instead of the customary unknown mixtures (Tyczkowski and Hamilton, 1986b). The objective of the present study was to investigate the progression of changes in tissue carotenoids of
2074
Downloaded from http://ps.oxfordjournals.org/ at Oakland University on June 3, 2015
ABSTRACT The progression of changes in carotenoid metabolism during pale-bird syndrome caused by a coccidial infection was investigated. Male broiler chickens 15 days of age on a yellow corn and soybean meal-based diet were infected with Eimeria acervulina oocysts and their serum, liver, and toe webs were sampled at 0,4, 6, and 10 days postinfection for HPLC analysis of carotenoids. At 4 days postinfection a drastic reduction (71%) in serum lutein, the main body carotenoid, and smaller reductions in liver (58%) and toe webs (38%) occurred. Derivative forms of lutein, mainly esters, continued to be lost from tissues for 10 days postinfection. These carotenoids were apparently lost via the intestinal tract because birds placed on a white corn and soybean meal-based diet at time of infection had lutein in their jejunal contents even at 7 days postinfection. The loss of carotenoids from the body was accompanied by a decreasing ability to absorb canthaxanthin, a red carotenoid, from the intestinal contents. The absorption of canthaxanthin measured at 0, 3, 4, 5, 6, and 7 days reached its low point of 1% of preinfection ability on Day 5 before a slow recovery commenced. Thus, the pale-bird syndrome caused by E. acervulina appeared to be the result of a loss of previously absorbed carotenoids coupled with drastic malabsorption of dietary carotenoids. (Key words: Eimeria acervulina, lutein, malabsorption, carotenoids, excretion)
CAROTENOID METABOLISM AND EIMERIA ACERVUUNA
chickens during infection with E. acervulina in order to determine whether the PBS caused by E. acervulina is the result of an efflux of oxycarotenoids from the serum and tissues, an inability to absorb oxycarotenoids from the intestinal tract, or a combination of both mechanisms. MATERIALS AND METHODS
Husbandry
Survey of Tissue Carotenoids During Infection At 0, 4, 6, and 10 days postinfection, four groups of four birds were bled (2.5 mL per bird) for serum collection, killed by cervical dislocation, and their livers and one foot were removed. The samples of each tissue were combined on a group basis and stored at -20 C prior to analysis. The toe webs of the feet after thawing were sampled with a circular (3 mm diameter) punch (Marusich and Bauernfeind, 1981) to provide four circles per foot. The livers were homogenized in deionized water (1:5, wt/vol).
3
Roxanthin Red 10, Hoffinann-LaRoche, Inc., Nulley, NJ 07110.
TABLE 1. Composition of diet Ingredients and calculated analysis Ground com1 Soybean meal (48.5% CP) Cottonseed oil Dicalcium phosphate (21% Ca, 18% P) Ground limestone Salt DL-methionine (98%) Vitamin premix Mineral premix Calculated analysis: Protein, % Fat, % ME, kcal per kg
Percentage 62.22 32.00 2.00 2.50 .60 .32 .26 .05 .05 21.00 4.50 3,041
1 The corn was yellow or white as specified for the experiment 2 Vitamin premix provided the following per kilogram of diet: vitamin A, 6,600 IU; cholecalciferol, 3,300 IU; vitamin E, 5.5 IU; niacin, 44 mg; pantothenic acid, 11 mg; riboflavin, 6.6 mg; menadione sodium bisulfite, 4.4 mg; pyridoxine, 1.1 mg; thiamine, 1.1 mg; folic acid, .22 mg; biotin, 28 Hg; vitamin Bj2, 13 Mg; selenium, .10 mg. 3 Mineral premix provided the following per kilogram of diet: Mn, 60 mg; Zn, 60 mg; Fe, 20 mg; Cu, 4.4 mg; I, .35 mg; Co, .05 mg.
Egress and Absorption of Carotenoids During Infection In another experiment, birds at 0, 3,4, and 5 days postinfection were removed from the yellow com and soybean meal-based diet. They were placed on a white corn and soybean mealbased diet (Table 1) essentially free of lutein (
