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Changes in Fowl Plasma α-Lipoproteins Caused by Starvation
STERCULIA
305
FOETIDA OIL AND HEMATOLOGY
effects of estrogen and androgen on osmotic fragility and fatty acid composition of erythrocytes in the chicken. Can. J. Physiol. Pharm. 44: 379-387. Mattern, C. G. T., F. S. Brackett and B. J. Olson, 1957. Determination of number and size of particles by electrical gating: blood cell. J. Appl. Physiol. 10: 56-70. Phelps, R. A., F. S. Shenstone, A. R. Kemmerer and R. J. Evans, 1965. A review of cyclopropenoid compounds: Biological effects of some derivatives. Poultry Sci. 44: 358-394. Reiser, R., and P. K. Raju, 1964. The inhibition of saturated fatty acid dehydrogenation by dietary
fat containing sterculic and malvalic acids. Biochem. Biophys. Res. Comm. 17: 8-11. Steel, R. G. D., and J. H. Torrie, 1960. Principles and Procedures of Statistics. McGraw-Hill Book Company, Inc., New York. Washburn, K. W., 1965. A genetic and physiological investigation of achromatosis., anemia, and double spurs occuring in a population of domestic fowl. Ph.D. Thesis, The University of Massachusetts, p. 170. Washburn, K. W., 1968. Effects of age of bird and hemoglobin type on the concentration of adult hemoglobin components of the domestic fowl. Poultry Sci. 47 : 1083-1089.
Changes in Fowl Plasma a-Lipoproteins Caused by Starvation R. W. BIDE Animal Pathology Division, Health of Animals Branch, Canada Department of Agriculture, Animal Diseases Research Institute (Western), P.O. Box 640, Lethbridge, Alberta, Canada (Received for publication June 10, 1971)
ABSTRACT Splitting and retardation of the a-lipoprotein bands on starch gel electrophoresis was observed in the plasmas of 15-week old birds starved for as little as 15 hours and in birds given reduced feed levels (10 gm./bird) for as little as four days. Feeding restored the normal condition within 3 hours. The changes were essentially the same as those produced by the addition of turpentines, acids or cationic soaps to plasma in vitro. These changes may provide a sensitive method for evaluating the extent of anorexia caused by disease or experimental conditions. POULTRY SCIENCE 51: 305-309,
1972
INTRODUCTION
MATERIALS AND METHODS
HE plasma a-lipoprotein of the chicken may be altered by the addition of soaps, turpentine and other compounds so that on starch gel electrophoresis the main a-lipoprotein band is "split and retarded" (Darcel et al., 1968) so that several slower bands are seen in place of the single main band (cf. Fig. 1). The physical change involved is probably a net reduction in the negative charges on or in the lipid micelles which may or may not cause a change in micellar structure but certainly results in a reduction in the electrophoretic mobility of the lipoproteins. This report describes similar changes in the avian plasma a-lipoproteins brought about by starvation.
Plasma was collected from heparinized blood drawn either by heart puncture using heparinized "Vacutainer" tubes (Beckton, Dickson Ltd., Rutherford, N.J.) or from wing veins using heparinized syringes. The plasma was stored for no more than 24 hrs. at — 20°C. in acrylic plastic vials before use. All birds were 15-week old White Leghorns of East Lansing Strain #151, which had been raised to 6 weeks of age on Ottawa Starter Ration and then maintained on Ottawa Intermediate Rearing Ration, both custom made by Master Feeds Division of Maple Leaf Milling Co., Calgary, Alberta. Equal numbers of males and fe-
T
306
R. W.
1
2
3
null'
BIDE
4
5
6
7
8
mm
wKm
«
IP
....
--—- mflii
• i <
A
FIG. 1. Changes in the a-lipoproteins caused by starvation and reduced feed intake. The gel patterns show plasma patterns from (1,5) normal birds; (2,3) 48-hours-starved birds; (4) bird starved 48 hours and fed, plasma sample obtained 4 hrs. after feeding; (6,7,8) birds on reduced feed for four, five, and six days, respectively.
males were present in each experimental group. Water was provided ad libitum to all birds. Starch gel electrophoresis was carried out at pH 8.6 using a discontinuous buffer system. The plasma samples were prestained for lipoproteins with Sudan Black and then stained for protein after electrophoresis with Buffalo Black. The relationships between bands and the identification of the bands in this system have been described in detail (Merriman and Darcel, 1964; Darcel et al., 1968). The extent of the a-lipoprotein changes were evaluated using a simple scoring system (Darcel et al, 1968). A full effect, value 2, was judged by the absence of the front-running a-lipoprotein band from its normal position, i.e. retarded so that no band was visible in that position (cf. Fig. 1-2, 3), whether or not splitting also occurred. A partial effect, value 1, was scored for any sample in which the fastest and major a-lipoprotein band was split and
the parts retarded resulting in a number of bands but always with some of the main band in the original position (cf. Fig. 1-6, 7, 8). Normal plasma, scored 0, contains 1 heavy and maybe two light bands (Fig. 1-1, 5) but seldom a slow band as well (Fig. 1-4). The accumulated score was divided by the maximum possible score and converted to a percent figure. In the starvation-recovery experiments, feed was withheld from 24 birds for 48 hours. Blood samples were taken from the wing veins before the food was withdrawn and after IS and 48 hours. After the blood samples were taken on the second day, the birds were fed and blood samples were taken each hour for 6 hours. At the end of this period, the birds were sacrificed. The plasma a-lipoproteins in each sample were examined by starch gel electrophoresis. In reduced-feed experiments, 24 birds were penned in groups of 3, all of one sex, and were given 30 gm. of feed per group each day for 10 days. This was about one
307
STARVATION AND PLASMA LIPOPROTEINS
• •
Origin
FIG. 2. Changes in the proteins caused by starvation and reduced feed intake. The gel patterns show plasma patterns from (1,5) normal birds; (2,3) 48 hours-starved birds; (4) bird starved 48 hours and fed, plasma sample obtained 4 hrs. after feeding; (6,7,8) birds on reduced feed for four, five, and six days, respectively.
sixth of the normal daily feed consumption of these birds. Blood samples were taken daily and the a-lipoproteins were examined by starch gel electrophoresis. RESULTS Starvation and Recovery. After 48 hours starvation, the a-lipoproteins in all birds showed the full effect, the main band was retarded with no material remaining in the original position. Bands had appeared near the origin and about ^ of the way up the gel. In some plasmas, the main band was diffuse (Fig. 1-2) and in others it appeared as a number of closer bands (Fig. 1-3). Overnight starvation, 15 hours, produced a 70% effect (Fig. 3) with more than half of the samples being fully affected. Two hours after feeding a number of the birds had "recovered" and three hours later, all birds had normal a-lipoproteins (Fig. 3). The protein bands reflect these same changes (Fig. 2).
Effect oj Reduced Feed. The enforced reduction of food intake to 10 gm./bird/day for 8 days produced the same a-lipoprotein
Hours After Feeding
FIG. 3. The effect of starvation on a-lipoproteins of chicken plasma. The birds were without feed for 48 hours and then fed. The percent lipoprotein effect is a rough measure of the number of birds affected and the extent of the effects as seen following starch gel electrophoresis.
308
R. W. BIDE
100 -
tt ©
^
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60 -
+» O
ft O
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/ /
a
/
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J
B.
f c» 0
* 1
* 2
" 3
i 4
i 0
i 6
i 7
i 8
9
Days Starvation
FIG. 4. The effect of reduced feed upon the plasma a-lipoproteins of chicken plasma. Birds, in groups of 3, were given 10 gm./bird/day of feed and the percent a-Iipoprotein effects studied by electrophoresis.
changes as starvation for 48 hours. The a-lipoprotein patterns showed no significant changes for 3 days (Fig. 4). Over the following days, the a-lipoprotein patterns change progressively until after 6 days the plasma from all the birds showed split and retarded a-lipoprotein bands. The transition between normal and the fully affected picture was prolonged so that for several days the majority of the plasma patterns were of the "partial effect" shown in Fig. 1-6, 7, 8. As the period of reduced feed was extended the effects increased until on the 8th day all were of the fully affected type shown in Fig. 1-2, 3. Similar changes were seen in the protein patterns (Fig. 2). DISCUSSION Starvation, for as little as 15 hours will cause marked changes in the a-lipoproteins in chicken plasma. The main a-lipoprotein band is split and retarded to varying degrees in a manner very similar to that seen when fowl plasma is treated with old turpentine, acids or cationic detergents (Darcel et al., 1968). These effects would appear
to be the result of changes in the net charge per mass of the lipoprotein micelles. The in vitro changes are probably caused by the addition of positive materials. In vivo the reduction of total negative charge would appear to be a likely mechanism to explain a similar change (Darcel, 1967; Bide and Darcel, 1969). In the case of the starved birds, the mechanisms by which these changes occur are unknown but there are several plausible explanations for the observed changes. There may be an increased utilization of the lipoproteins in compensation for the drop in nutrients. Equally a reduced lipoprotein synthesis may be the cause. Reductions in plasma inorganic phosphate have been reported in birds on either reduced feed or phosphate (Gardiner, 1962) and as there is a large daily requirement for choline (Bird et al., 1966), the absence of either dietary factor may result in reduced synthesis of phospholipid. The position and nature of the plasma a-lipoprotein bands on starch gel would appear to be a sensitive indicator of the feeding state of the chicken. Feeding about one sixth the normal daily feed intake caused changes in the plasma a-lipoprotein picture (Fig. 4). In addition, feeding a starved bird resulted in an essentially normal a-lipoprotein picture in as little as 2 hours in some cases and in all cases after 3 hours' following a 48 hour fast. Over longer periods on a reduced feed intake, the a-lipoprotein bands were altered in a similar way. Although these studies are of a preliminary nature and leave many questions unanswered, several points which arise should be stressed. In human clinical chemistry, blood samples for analysis are often taken after an overnight fast. If a similar practice were to be followed with the chicken, a number of confusing half changes and many erroneous observations would result from the alterations in the a-lipoproteins described in this report. Similarly, in disease conditions in which the feed intake is reduced,
STARVATION AND PLASMA LIPOPROTEINS
there may be changes in the a-lipoproteins which are the result of the anorexia rather than the progress of the disease state. On the positive side, the changes observed were sensitive to changes in feed intake and so may be useful in the evaluation of dietary factors in relation to feed intake on different diets and as a measure of anorexia attendant to disease states. ACKNOWLEDGMENTS The author wishes to thank Mr. H. Klassen, Mr. S. McGee and Mrs. Norma Dow for their technical assistance in this work. Thanks are also due to Dr. E. Gardiner of Canada Agriculture Research Station, Lethbridge and Dr. S. Magwood for their help with the manuscript. The photographic work was done by Mr. N. Kloppenborg and staff of the Photograph Section, Canada Agriculture Research Station, Lethbridge.
309
REFERENCES Bide, R. W., and C. le Q. Darcel, 1969. Changes in the phosphorus in the plasma fractions in avian erythroblastosis. Poultry Sci. 48: 795-798. Bird, H. R., H. J. Almquist, D. R. Clandinin, W. W. Cravens, F. W. Hill and J. McGinnis. Nutrient Requirements of Poultry. 5th Ed. Publication 1345 of Nat. Acad. Sci., National Research Council, Wash., D.C., p. 4, 20. Darcel, C. le Q., 1967. Reduction in the concentration of phosphatidyl serine in the plasma of birds with avian erythroblastosis. Nature, 215: 647-648. Darcel, C. le Q., R. W. Bide and M. Merriman, 1968. Properties of turpentine relative to the effects on fowl plasma in vitro. I. The simulation of "leukemic" changes in normal plasma. Can. J. Biochem. 46: 503-508. Gardiner, E., 1962. The relationship between dietary phosphorus level and the level of plasma inorganic phosphorus in chicks. Poultry Sci. 4 1 : 1156-1163. Merriman, M., and C. le Q. Darcel, 1964. Confirmation of plasma protein changes in avian erythroblastosis. Can. J. Biochem. 42: 293-297.
Toxigenic Fungi from Poultry Feed and Litter JOSEPH LOVETT U. S. Department of Health, Education, and Welfare, Public Health Service, Food and Drug Administration, Division of Microbiology, Cincinnati, Ohio 45226 (Received for publication June 11, 1971) ABSTRACT Fungi isolated from feed and litter of two Ohio poultry farms were screened for toxin production. Fourteen-day-slant cultures were used to inoculate neopeptone dextrose, Czapek-Dox, and Mycological broth media. Four-day chick embryos were inoculated with 0.2 ml. of culture filtrate via the air cell. Embryo death at 9 days was used as the toxicity indicator. Those fungi found toxigenic in one or more media were Aspergillus chevalieri (one), A. fumigatus (one), A. terreus (two), Pmicillium cyclopium (five), P. patulum (two), and one each Fusarium and Scopulariopsis sp. Of those isolates screened 13% were found toxigenic. POULTRY SCIENCE 51:
INTRODUCTION
M
YCOTOXINS in foodstuffs are recognized as a public health problem of considerable importance. Fungal toxins in feed and foods have become a major research area since the 1961 discovery of the carcinogenicity of the aflatoxins. Hundreds
309-313,
1972
of species of more than a dozen fungal genera are known to be toxigenic. Poultry mycotoxicosis results from ingestion by poultry of toxic fungal metabolites in feed, litter, and water into which feed is spilled (Forgacs et al., 1962; Forgacs, 1966; Abrams, 1965). Some fungi
305
FOETIDA OIL AND HEMATOLOGY
effects of estrogen and androgen on osmotic fragility and fatty acid composition of erythrocytes in the chicken. Can. J. Physiol. Pharm. 44: 379-387. Mattern, C. G. T., F. S. Brackett and B. J. Olson, 1957. Determination of number and size of particles by electrical gating: blood cell. J. Appl. Physiol. 10: 56-70. Phelps, R. A., F. S. Shenstone, A. R. Kemmerer and R. J. Evans, 1965. A review of cyclopropenoid compounds: Biological effects of some derivatives. Poultry Sci. 44: 358-394. Reiser, R., and P. K. Raju, 1964. The inhibition of saturated fatty acid dehydrogenation by dietary
fat containing sterculic and malvalic acids. Biochem. Biophys. Res. Comm. 17: 8-11. Steel, R. G. D., and J. H. Torrie, 1960. Principles and Procedures of Statistics. McGraw-Hill Book Company, Inc., New York. Washburn, K. W., 1965. A genetic and physiological investigation of achromatosis., anemia, and double spurs occuring in a population of domestic fowl. Ph.D. Thesis, The University of Massachusetts, p. 170. Washburn, K. W., 1968. Effects of age of bird and hemoglobin type on the concentration of adult hemoglobin components of the domestic fowl. Poultry Sci. 47 : 1083-1089.
Changes in Fowl Plasma a-Lipoproteins Caused by Starvation R. W. BIDE Animal Pathology Division, Health of Animals Branch, Canada Department of Agriculture, Animal Diseases Research Institute (Western), P.O. Box 640, Lethbridge, Alberta, Canada (Received for publication June 10, 1971)
ABSTRACT Splitting and retardation of the a-lipoprotein bands on starch gel electrophoresis was observed in the plasmas of 15-week old birds starved for as little as 15 hours and in birds given reduced feed levels (10 gm./bird) for as little as four days. Feeding restored the normal condition within 3 hours. The changes were essentially the same as those produced by the addition of turpentines, acids or cationic soaps to plasma in vitro. These changes may provide a sensitive method for evaluating the extent of anorexia caused by disease or experimental conditions. POULTRY SCIENCE 51: 305-309,
1972
INTRODUCTION
MATERIALS AND METHODS
HE plasma a-lipoprotein of the chicken may be altered by the addition of soaps, turpentine and other compounds so that on starch gel electrophoresis the main a-lipoprotein band is "split and retarded" (Darcel et al., 1968) so that several slower bands are seen in place of the single main band (cf. Fig. 1). The physical change involved is probably a net reduction in the negative charges on or in the lipid micelles which may or may not cause a change in micellar structure but certainly results in a reduction in the electrophoretic mobility of the lipoproteins. This report describes similar changes in the avian plasma a-lipoproteins brought about by starvation.
Plasma was collected from heparinized blood drawn either by heart puncture using heparinized "Vacutainer" tubes (Beckton, Dickson Ltd., Rutherford, N.J.) or from wing veins using heparinized syringes. The plasma was stored for no more than 24 hrs. at — 20°C. in acrylic plastic vials before use. All birds were 15-week old White Leghorns of East Lansing Strain #151, which had been raised to 6 weeks of age on Ottawa Starter Ration and then maintained on Ottawa Intermediate Rearing Ration, both custom made by Master Feeds Division of Maple Leaf Milling Co., Calgary, Alberta. Equal numbers of males and fe-
T
306
R. W.
1
2
3
null'
BIDE
4
5
6
7
8
mm
wKm
«
IP
....
--—- mflii
• i <
A
FIG. 1. Changes in the a-lipoproteins caused by starvation and reduced feed intake. The gel patterns show plasma patterns from (1,5) normal birds; (2,3) 48-hours-starved birds; (4) bird starved 48 hours and fed, plasma sample obtained 4 hrs. after feeding; (6,7,8) birds on reduced feed for four, five, and six days, respectively.
males were present in each experimental group. Water was provided ad libitum to all birds. Starch gel electrophoresis was carried out at pH 8.6 using a discontinuous buffer system. The plasma samples were prestained for lipoproteins with Sudan Black and then stained for protein after electrophoresis with Buffalo Black. The relationships between bands and the identification of the bands in this system have been described in detail (Merriman and Darcel, 1964; Darcel et al., 1968). The extent of the a-lipoprotein changes were evaluated using a simple scoring system (Darcel et al, 1968). A full effect, value 2, was judged by the absence of the front-running a-lipoprotein band from its normal position, i.e. retarded so that no band was visible in that position (cf. Fig. 1-2, 3), whether or not splitting also occurred. A partial effect, value 1, was scored for any sample in which the fastest and major a-lipoprotein band was split and
the parts retarded resulting in a number of bands but always with some of the main band in the original position (cf. Fig. 1-6, 7, 8). Normal plasma, scored 0, contains 1 heavy and maybe two light bands (Fig. 1-1, 5) but seldom a slow band as well (Fig. 1-4). The accumulated score was divided by the maximum possible score and converted to a percent figure. In the starvation-recovery experiments, feed was withheld from 24 birds for 48 hours. Blood samples were taken from the wing veins before the food was withdrawn and after IS and 48 hours. After the blood samples were taken on the second day, the birds were fed and blood samples were taken each hour for 6 hours. At the end of this period, the birds were sacrificed. The plasma a-lipoproteins in each sample were examined by starch gel electrophoresis. In reduced-feed experiments, 24 birds were penned in groups of 3, all of one sex, and were given 30 gm. of feed per group each day for 10 days. This was about one
307
STARVATION AND PLASMA LIPOPROTEINS
• •
Origin
FIG. 2. Changes in the proteins caused by starvation and reduced feed intake. The gel patterns show plasma patterns from (1,5) normal birds; (2,3) 48 hours-starved birds; (4) bird starved 48 hours and fed, plasma sample obtained 4 hrs. after feeding; (6,7,8) birds on reduced feed for four, five, and six days, respectively.
sixth of the normal daily feed consumption of these birds. Blood samples were taken daily and the a-lipoproteins were examined by starch gel electrophoresis. RESULTS Starvation and Recovery. After 48 hours starvation, the a-lipoproteins in all birds showed the full effect, the main band was retarded with no material remaining in the original position. Bands had appeared near the origin and about ^ of the way up the gel. In some plasmas, the main band was diffuse (Fig. 1-2) and in others it appeared as a number of closer bands (Fig. 1-3). Overnight starvation, 15 hours, produced a 70% effect (Fig. 3) with more than half of the samples being fully affected. Two hours after feeding a number of the birds had "recovered" and three hours later, all birds had normal a-lipoproteins (Fig. 3). The protein bands reflect these same changes (Fig. 2).
Effect oj Reduced Feed. The enforced reduction of food intake to 10 gm./bird/day for 8 days produced the same a-lipoprotein
Hours After Feeding
FIG. 3. The effect of starvation on a-lipoproteins of chicken plasma. The birds were without feed for 48 hours and then fed. The percent lipoprotein effect is a rough measure of the number of birds affected and the extent of the effects as seen following starch gel electrophoresis.
308
R. W. BIDE
100 -
tt ©
^
•
i
i 10
/ /
60 -
+» O
ft O
/ /
P<
/
3 «•"
/ /
/ /
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I 20 -
J
B.
f c» 0
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" 3
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i 0
i 6
i 7
i 8
9
Days Starvation
FIG. 4. The effect of reduced feed upon the plasma a-lipoproteins of chicken plasma. Birds, in groups of 3, were given 10 gm./bird/day of feed and the percent a-Iipoprotein effects studied by electrophoresis.
changes as starvation for 48 hours. The a-lipoprotein patterns showed no significant changes for 3 days (Fig. 4). Over the following days, the a-lipoprotein patterns change progressively until after 6 days the plasma from all the birds showed split and retarded a-lipoprotein bands. The transition between normal and the fully affected picture was prolonged so that for several days the majority of the plasma patterns were of the "partial effect" shown in Fig. 1-6, 7, 8. As the period of reduced feed was extended the effects increased until on the 8th day all were of the fully affected type shown in Fig. 1-2, 3. Similar changes were seen in the protein patterns (Fig. 2). DISCUSSION Starvation, for as little as 15 hours will cause marked changes in the a-lipoproteins in chicken plasma. The main a-lipoprotein band is split and retarded to varying degrees in a manner very similar to that seen when fowl plasma is treated with old turpentine, acids or cationic detergents (Darcel et al., 1968). These effects would appear
to be the result of changes in the net charge per mass of the lipoprotein micelles. The in vitro changes are probably caused by the addition of positive materials. In vivo the reduction of total negative charge would appear to be a likely mechanism to explain a similar change (Darcel, 1967; Bide and Darcel, 1969). In the case of the starved birds, the mechanisms by which these changes occur are unknown but there are several plausible explanations for the observed changes. There may be an increased utilization of the lipoproteins in compensation for the drop in nutrients. Equally a reduced lipoprotein synthesis may be the cause. Reductions in plasma inorganic phosphate have been reported in birds on either reduced feed or phosphate (Gardiner, 1962) and as there is a large daily requirement for choline (Bird et al., 1966), the absence of either dietary factor may result in reduced synthesis of phospholipid. The position and nature of the plasma a-lipoprotein bands on starch gel would appear to be a sensitive indicator of the feeding state of the chicken. Feeding about one sixth the normal daily feed intake caused changes in the plasma a-lipoprotein picture (Fig. 4). In addition, feeding a starved bird resulted in an essentially normal a-lipoprotein picture in as little as 2 hours in some cases and in all cases after 3 hours' following a 48 hour fast. Over longer periods on a reduced feed intake, the a-lipoprotein bands were altered in a similar way. Although these studies are of a preliminary nature and leave many questions unanswered, several points which arise should be stressed. In human clinical chemistry, blood samples for analysis are often taken after an overnight fast. If a similar practice were to be followed with the chicken, a number of confusing half changes and many erroneous observations would result from the alterations in the a-lipoproteins described in this report. Similarly, in disease conditions in which the feed intake is reduced,
STARVATION AND PLASMA LIPOPROTEINS
there may be changes in the a-lipoproteins which are the result of the anorexia rather than the progress of the disease state. On the positive side, the changes observed were sensitive to changes in feed intake and so may be useful in the evaluation of dietary factors in relation to feed intake on different diets and as a measure of anorexia attendant to disease states. ACKNOWLEDGMENTS The author wishes to thank Mr. H. Klassen, Mr. S. McGee and Mrs. Norma Dow for their technical assistance in this work. Thanks are also due to Dr. E. Gardiner of Canada Agriculture Research Station, Lethbridge and Dr. S. Magwood for their help with the manuscript. The photographic work was done by Mr. N. Kloppenborg and staff of the Photograph Section, Canada Agriculture Research Station, Lethbridge.
309
REFERENCES Bide, R. W., and C. le Q. Darcel, 1969. Changes in the phosphorus in the plasma fractions in avian erythroblastosis. Poultry Sci. 48: 795-798. Bird, H. R., H. J. Almquist, D. R. Clandinin, W. W. Cravens, F. W. Hill and J. McGinnis. Nutrient Requirements of Poultry. 5th Ed. Publication 1345 of Nat. Acad. Sci., National Research Council, Wash., D.C., p. 4, 20. Darcel, C. le Q., 1967. Reduction in the concentration of phosphatidyl serine in the plasma of birds with avian erythroblastosis. Nature, 215: 647-648. Darcel, C. le Q., R. W. Bide and M. Merriman, 1968. Properties of turpentine relative to the effects on fowl plasma in vitro. I. The simulation of "leukemic" changes in normal plasma. Can. J. Biochem. 46: 503-508. Gardiner, E., 1962. The relationship between dietary phosphorus level and the level of plasma inorganic phosphorus in chicks. Poultry Sci. 4 1 : 1156-1163. Merriman, M., and C. le Q. Darcel, 1964. Confirmation of plasma protein changes in avian erythroblastosis. Can. J. Biochem. 42: 293-297.
Toxigenic Fungi from Poultry Feed and Litter JOSEPH LOVETT U. S. Department of Health, Education, and Welfare, Public Health Service, Food and Drug Administration, Division of Microbiology, Cincinnati, Ohio 45226 (Received for publication June 11, 1971) ABSTRACT Fungi isolated from feed and litter of two Ohio poultry farms were screened for toxin production. Fourteen-day-slant cultures were used to inoculate neopeptone dextrose, Czapek-Dox, and Mycological broth media. Four-day chick embryos were inoculated with 0.2 ml. of culture filtrate via the air cell. Embryo death at 9 days was used as the toxicity indicator. Those fungi found toxigenic in one or more media were Aspergillus chevalieri (one), A. fumigatus (one), A. terreus (two), Pmicillium cyclopium (five), P. patulum (two), and one each Fusarium and Scopulariopsis sp. Of those isolates screened 13% were found toxigenic. POULTRY SCIENCE 51:
INTRODUCTION
M
YCOTOXINS in foodstuffs are recognized as a public health problem of considerable importance. Fungal toxins in feed and foods have become a major research area since the 1961 discovery of the carcinogenicity of the aflatoxins. Hundreds
309-313,
1972
of species of more than a dozen fungal genera are known to be toxigenic. Poultry mycotoxicosis results from ingestion by poultry of toxic fungal metabolites in feed, litter, and water into which feed is spilled (Forgacs et al., 1962; Forgacs, 1966; Abrams, 1965). Some fungi