Pathological Changes in Chickens, Ducks and Turkeys Fed High Levels of Rapeseed Oil C. Ratanasethkul, C. Riddell, R. E. Salmon and J. B. O'Neil*
ABSTRACT Rations containing 25% of either regular rapeseed oil (36% erucic acid), Oro rapeseed oil (1.9% erucic acid), soybean oil or a mixture of lard and corn oil were fed to chickens, ducks and turkeys. The regular rapeseed oil ration caused growth depression, increased feed conversion and anemia in all species. All the ducks and some of the chickens fed the regular rapeseed oil ration died. These dead birds were affected with hydropericardium and ascites. No deaths in the turkeys could be attributed to the regular rapeseed oil ration but some turkeys fed this ration had degenerative foci characterized by infiltrations of histiocytic and giant cells in the myocardium. Severe fatty change in the heart, skeletal muscles, spleen and kidney was found at an early age in all birds fed the regular rapeseed oil ration. Less severe fatty change but no other lesions were found in birds fed the Oro rapeseed oil and soybean oil rations.
RESUME On a servi 'a des poulets, des canards et des dindes, des rations contenant 25% d'huile de colza reguliere (36% d'acide erucique) ou Oro (1.9% d'acide erucique), de l'huile de soja ou un melange de lard et d'huile de mais. L'huile de colza reguliere provoqua un retard de croissance, une augmentation de la conversion
*Western College of Veterinary Medicine (Ratanasethakul and Riddeli)), Department of Poultry Science (O'Neil), University of Saskatchewan, Saskatoon, Saskatchewan and Canada Department of Agriculture Research Station (Salmon), Swift Current, Saskatchewan. This study was supported by the Rapeseed Utilization Assistance Program of the Rapeseed Association of Canada. Based on a thesis submitted by the senior author to the University of Saskatchewan in partial fulfilment of the requirements for the Master of Science degree in Veterinary Pathology. Submitted January 20, 1976.
360
alimentaire et de l'anemie, chez toutes les especes. Tous les canards et certains des poulets nourris avec la ration contenant de l'huile de colza reguliere moururent. Tous ces oiseaux presentaient un hydropericarde et de l'ascite. Cette ration ne tua pas de dindes, mais le myocarde d'un certain nombre d'entre eux contenait des foyers de degenerescence, caracterises par une infiltration d'histiocytes et de cellules geantes. Le coeur, les muscles squelettiques, la rate et les reins de tous les oiseaux nourris avec la ration contenant de l'huile de colza reguliere developperent rapidement une metamorphose graisseuse marqu&e. Les oiseaux nourris avec les rations contenant de l'huile de colza Oro ou de l'huile de soya, ne developperent pas d'autre lesion qu'un metamorphose graisseuse moins marquee.
INTRODUCTION Rapeseed oil fed at levels of 30-60 cal % in the diet of White Peking ducks produced growth retardation, mortality, increased packed cell volume and reticulocyte count, severe hydropericardium, hypertrophy of heart, cirrhotic changes of the liver, lipidosis of the spleen and fatty infiltration associated with cellular infiltration in the heart and skeletal muscles. The rapeseed oil contained 40% erucic acid and the growth depression and lesions were reproduced by feeding ducks a ration containing glyceryl trierucate (2). Growth depression and fatty infiltration in the myocardium were noted in turkeys fed a diet including 20% rapeseed oil which contained 35% erucic acid. Mortality, hydropericardium and cirrhotic changes in the liver did not occur in these turkeys but focal lesions in the myocardium consisting of necrotic cellular debris, giant cells, histiocytes and heterophils were observed (Salmon, R. E. and C. Riddell, unpublished data, 1971).
Can. J. comp. Med.
The following experiment was designed to study this apparent difference in response of different avian species to high dietary levels of rapeseed oil.
reading the total protein value (TP) which was expressed in g/100 ml of plasma. A blood smear was made from a small drop
of blood from the heparinized capillary tube and stained with Wright-Giemsa stain.
MATERIALS AND METHODS EXPERIMENTAL BIRDS AND RATIONS Thirty-two male and 32 female day old chickens of a commercial broiler chicken strain, 60 mixed sex day old White Peking ducks and 32 male and 32 female day old turkeys of a commercial Broad Breasted White turkey strain were brooded using infrared lamps in 24 floor pens. Each pen contained four male and four female chickens, seven to eight unsexed ducklings or four male and four female turkeys. Unlimited feed was provided for all birds from one to fifty-two days of age at which time the experiment was terminated. Four different rations were used and each ration was fed to two pens of birds of each species. The rations varied only in the type of oil added as outlined in Table I. Ration 1, designed as a control contained 18.75% lard and 6.25% corn oil; ration 2, 25% soybean oil (SBO); ration 3, 25% Oro rapeseed oil (ORSO) ration 4, 25% regular rapeseed oil (RRSO). The ORSO contained 1.9% erucic acid while the RRSO contained 36% erucic acid. Fatty acid analysis of oils and rations was carried out as described previously (6) and results are tabulated in Table II. Birds were weighed weekly and feed consumption recorded. HEMATOLOGICAL EXAMINATION A heparinized microcapillary tube' was filled directly from a punctured vessel. The sealed capillary tubes were centrifuged2 at 11,000 rpm for five minutes. The packed cell volume (PCV) in the capillary tube was determined with a microcapillary reader3 and expressed in percent. A single drop of plasma from the capillary tube was pressed on the prism of a TS meter4 for "'Capilets" heparinized microhematocrit capillary tubes, 75 mm long and 1.0 mm diameter, by Canlab, U.S.A.
2lnternational Micro-capillary Centrifuge Model MB, international Equipment Company, Needham, Massachusetts. 3International Micro-capillary Reader Model CR, International Equipment Company, Needham, Massachusetts. 4TS meter Series TS-B, American Optical Company, Keene, New Hampshire.
Volume 40
-
October, 1976
PATHOLOGICAL EXAMINATION
All birds which died during the experiment were necropsied. One bird of each sex when the sex was known was selected randomly from each pen and necropsied at seven and 28 days. The remaining birds were necropsied at 52 days. Hearts, livers and spleens were weighed. Sections of heart, lung, liver, kidney, spleen, pancreas, duodenum, brailn and superficial pectoral and adductor muscles were collected in 10% formalin for histopathology. These tissues were embedded in paraffin and sectioned at 7 ,.u and stained with hematoxylin and eosin. Frozen TABLE I. Composition (percent) of Control, 25%C Soybean oil, 25%o Oro Rapeseed Oil and 25%`c, Regular Rapeseed Oil Rations for Chickens, Ducks and Turkeys (0-52 days) Ingredient Soybean meal (48%7o protein)...... Herring fish meal (73% protein).. Dried brewers yeast.............. Alphafloc...................... Dehydrated alfalfa............... Dicalcium phosphate............. Calcium carbonate............... Salt............................ Premixa........................ DL-methionine (99%c)............ Oil (variable)b .................. Total ......................
c
52.750 10.000 2.000 3.000 2.000 3.250 1.000 0.250 0.443 0.250 25.000 99.943
Calculated analyss Protein % ....................... 36.30 Metabolizable energy (K cal/kg)c. 3982.00 C/P ratio....................... 110.00 Calcium % ..................... 1.57 Phosphorus % .................. 0.99 Methionine and cystine %cc........ 1.32 Lysine %O....................... 2.16 aPremix contributes, per kg of ration: vit. A, 15,840 I.U.; vit. D3, 3,300 I.U.: vit. E, 26.4 I.U.; vit. B12, 15.8 mcg; riboflavin, 10.8 mg; calcium-d-pantothenate, 15.8 mg; niacin, 90 mg; pyridoxine, 3.6 mg; choline chloride, 720 mg; folic acid, 1.32 mg; menadione sodium bisulphite, 5.28 mg; biotin, 198 mcg; ethoxyquin, 150 mg; amprolium, 150 mg; trace mineral mix, 600 mg bRation 1, 18.75% lard 6.25% com oil; ration 2, 25%to soybean oil; ration 3, 25%o Oro rapeseed oil; ration 4, 25% regular rapeseed oil. Soybean and rapeseed oils were fully refined, bleached and deodorized cOil contributed approximately 58% of metabolizable calories, varying slightly with oil and species
361
TABLE II. Fatty Acid Composition of Oils and Control (#1), 25% Soybean Oil (#2), 25% Oro Rapeseed Oil (#3), and 25% Regular Rapeseed Oil (#4) Rationsa Fatty acid 14:0 16:0 16:1 16:2 18:0 18:1 18:2 18:3 20:1 22:1
Lard 1.5 24.8 3.5 0.5 13.6 45.0 9.9 0.5 0.7
Corn oil
Soybean oil
Oro rapeseed oil
Regular rapeseed oil
10.6
9.9
4.6
3.4
1.5 25.1 60.5 1.9 0.5
4.0 25.7 52.1 7.6 0.7
1.6 58.6 22.6 8.5 2.4 1.9
1.2 21.2 17.4 7.5 13.3 36.1
Mixed rations #3 #1 #2 1.7 0.4 0.3 20.6 10.4 5.9 3.0 0.5 1.7 9.5 4.0 36.7 25.1 54.1 24.4 50.4 24.7 1.7 7.3 8.0 1.1 0.9 2.4 1.6
#4 0.2 4.2 0.5
1.4 21.0 19.8 7.4 12.0 32.2
aExpressed as percent of methyl esters sections of selected tissues were stained with Oil Red 0 using Lillie and Ashburn's method (7).
STATISTICAL EVALUATION Body weights, feed conversions, relative weights of organs and blood values were subjected to analysis of variance and the means compared by Duncan's multiple range test (10).
RESULTS BODY WEIGHTS AND FEED CONVERSION
TABLE III. Average Body Weight (g) and Feed Conversion (g feed/g body weight gained) of Chickens, Ducks and Turkeys Fed Control, 25% Soybean Oil (SBO), 25% Oro Rapeseed Oil (ORSO) and 25% Regular Rapeseed Oil (RRSO) Rations
Body weight at 28
at 49
0-49
days
days
days
724.3a
1522.48
1.47a
667.5a
1430.98
1.74b
467.7b
1058.4b
1.67b
Ducks Control 25% SBO 25% ORSO.
1655.5a
2871.7a
1532.5a
2715.08
1.88a
1582.58
2831.3a
2.29b
275.5b
-c
c
Turkeys Control .... 257o SBO .. 25%o ORSO. 25% RRSO.
819.1a 758.4a 746.3a
2383.2a 2241.98 2035.7a
1.318 1.36a
Chickens Control
25% SBO 25%o ORSO. 25% RRSO.
25%o RRSO.
No significant differences were noted among the average body weights of the birds of the same species fed the control, SBO or ORSO rations throughout the experiment. The average body weights of all species fed the RRSO ration were significantly less (P
ABSTRACT Rations containing 25% of either regular rapeseed oil (36% erucic acid), Oro rapeseed oil (1.9% erucic acid), soybean oil or a mixture of lard and corn oil were fed to chickens, ducks and turkeys. The regular rapeseed oil ration caused growth depression, increased feed conversion and anemia in all species. All the ducks and some of the chickens fed the regular rapeseed oil ration died. These dead birds were affected with hydropericardium and ascites. No deaths in the turkeys could be attributed to the regular rapeseed oil ration but some turkeys fed this ration had degenerative foci characterized by infiltrations of histiocytic and giant cells in the myocardium. Severe fatty change in the heart, skeletal muscles, spleen and kidney was found at an early age in all birds fed the regular rapeseed oil ration. Less severe fatty change but no other lesions were found in birds fed the Oro rapeseed oil and soybean oil rations.
RESUME On a servi 'a des poulets, des canards et des dindes, des rations contenant 25% d'huile de colza reguliere (36% d'acide erucique) ou Oro (1.9% d'acide erucique), de l'huile de soja ou un melange de lard et d'huile de mais. L'huile de colza reguliere provoqua un retard de croissance, une augmentation de la conversion
*Western College of Veterinary Medicine (Ratanasethakul and Riddeli)), Department of Poultry Science (O'Neil), University of Saskatchewan, Saskatoon, Saskatchewan and Canada Department of Agriculture Research Station (Salmon), Swift Current, Saskatchewan. This study was supported by the Rapeseed Utilization Assistance Program of the Rapeseed Association of Canada. Based on a thesis submitted by the senior author to the University of Saskatchewan in partial fulfilment of the requirements for the Master of Science degree in Veterinary Pathology. Submitted January 20, 1976.
360
alimentaire et de l'anemie, chez toutes les especes. Tous les canards et certains des poulets nourris avec la ration contenant de l'huile de colza reguliere moururent. Tous ces oiseaux presentaient un hydropericarde et de l'ascite. Cette ration ne tua pas de dindes, mais le myocarde d'un certain nombre d'entre eux contenait des foyers de degenerescence, caracterises par une infiltration d'histiocytes et de cellules geantes. Le coeur, les muscles squelettiques, la rate et les reins de tous les oiseaux nourris avec la ration contenant de l'huile de colza reguliere developperent rapidement une metamorphose graisseuse marqu&e. Les oiseaux nourris avec les rations contenant de l'huile de colza Oro ou de l'huile de soya, ne developperent pas d'autre lesion qu'un metamorphose graisseuse moins marquee.
INTRODUCTION Rapeseed oil fed at levels of 30-60 cal % in the diet of White Peking ducks produced growth retardation, mortality, increased packed cell volume and reticulocyte count, severe hydropericardium, hypertrophy of heart, cirrhotic changes of the liver, lipidosis of the spleen and fatty infiltration associated with cellular infiltration in the heart and skeletal muscles. The rapeseed oil contained 40% erucic acid and the growth depression and lesions were reproduced by feeding ducks a ration containing glyceryl trierucate (2). Growth depression and fatty infiltration in the myocardium were noted in turkeys fed a diet including 20% rapeseed oil which contained 35% erucic acid. Mortality, hydropericardium and cirrhotic changes in the liver did not occur in these turkeys but focal lesions in the myocardium consisting of necrotic cellular debris, giant cells, histiocytes and heterophils were observed (Salmon, R. E. and C. Riddell, unpublished data, 1971).
Can. J. comp. Med.
The following experiment was designed to study this apparent difference in response of different avian species to high dietary levels of rapeseed oil.
reading the total protein value (TP) which was expressed in g/100 ml of plasma. A blood smear was made from a small drop
of blood from the heparinized capillary tube and stained with Wright-Giemsa stain.
MATERIALS AND METHODS EXPERIMENTAL BIRDS AND RATIONS Thirty-two male and 32 female day old chickens of a commercial broiler chicken strain, 60 mixed sex day old White Peking ducks and 32 male and 32 female day old turkeys of a commercial Broad Breasted White turkey strain were brooded using infrared lamps in 24 floor pens. Each pen contained four male and four female chickens, seven to eight unsexed ducklings or four male and four female turkeys. Unlimited feed was provided for all birds from one to fifty-two days of age at which time the experiment was terminated. Four different rations were used and each ration was fed to two pens of birds of each species. The rations varied only in the type of oil added as outlined in Table I. Ration 1, designed as a control contained 18.75% lard and 6.25% corn oil; ration 2, 25% soybean oil (SBO); ration 3, 25% Oro rapeseed oil (ORSO) ration 4, 25% regular rapeseed oil (RRSO). The ORSO contained 1.9% erucic acid while the RRSO contained 36% erucic acid. Fatty acid analysis of oils and rations was carried out as described previously (6) and results are tabulated in Table II. Birds were weighed weekly and feed consumption recorded. HEMATOLOGICAL EXAMINATION A heparinized microcapillary tube' was filled directly from a punctured vessel. The sealed capillary tubes were centrifuged2 at 11,000 rpm for five minutes. The packed cell volume (PCV) in the capillary tube was determined with a microcapillary reader3 and expressed in percent. A single drop of plasma from the capillary tube was pressed on the prism of a TS meter4 for "'Capilets" heparinized microhematocrit capillary tubes, 75 mm long and 1.0 mm diameter, by Canlab, U.S.A.
2lnternational Micro-capillary Centrifuge Model MB, international Equipment Company, Needham, Massachusetts. 3International Micro-capillary Reader Model CR, International Equipment Company, Needham, Massachusetts. 4TS meter Series TS-B, American Optical Company, Keene, New Hampshire.
Volume 40
-
October, 1976
PATHOLOGICAL EXAMINATION
All birds which died during the experiment were necropsied. One bird of each sex when the sex was known was selected randomly from each pen and necropsied at seven and 28 days. The remaining birds were necropsied at 52 days. Hearts, livers and spleens were weighed. Sections of heart, lung, liver, kidney, spleen, pancreas, duodenum, brailn and superficial pectoral and adductor muscles were collected in 10% formalin for histopathology. These tissues were embedded in paraffin and sectioned at 7 ,.u and stained with hematoxylin and eosin. Frozen TABLE I. Composition (percent) of Control, 25%C Soybean oil, 25%o Oro Rapeseed Oil and 25%`c, Regular Rapeseed Oil Rations for Chickens, Ducks and Turkeys (0-52 days) Ingredient Soybean meal (48%7o protein)...... Herring fish meal (73% protein).. Dried brewers yeast.............. Alphafloc...................... Dehydrated alfalfa............... Dicalcium phosphate............. Calcium carbonate............... Salt............................ Premixa........................ DL-methionine (99%c)............ Oil (variable)b .................. Total ......................
c
52.750 10.000 2.000 3.000 2.000 3.250 1.000 0.250 0.443 0.250 25.000 99.943
Calculated analyss Protein % ....................... 36.30 Metabolizable energy (K cal/kg)c. 3982.00 C/P ratio....................... 110.00 Calcium % ..................... 1.57 Phosphorus % .................. 0.99 Methionine and cystine %cc........ 1.32 Lysine %O....................... 2.16 aPremix contributes, per kg of ration: vit. A, 15,840 I.U.; vit. D3, 3,300 I.U.: vit. E, 26.4 I.U.; vit. B12, 15.8 mcg; riboflavin, 10.8 mg; calcium-d-pantothenate, 15.8 mg; niacin, 90 mg; pyridoxine, 3.6 mg; choline chloride, 720 mg; folic acid, 1.32 mg; menadione sodium bisulphite, 5.28 mg; biotin, 198 mcg; ethoxyquin, 150 mg; amprolium, 150 mg; trace mineral mix, 600 mg bRation 1, 18.75% lard 6.25% com oil; ration 2, 25%to soybean oil; ration 3, 25%o Oro rapeseed oil; ration 4, 25% regular rapeseed oil. Soybean and rapeseed oils were fully refined, bleached and deodorized cOil contributed approximately 58% of metabolizable calories, varying slightly with oil and species
361
TABLE II. Fatty Acid Composition of Oils and Control (#1), 25% Soybean Oil (#2), 25% Oro Rapeseed Oil (#3), and 25% Regular Rapeseed Oil (#4) Rationsa Fatty acid 14:0 16:0 16:1 16:2 18:0 18:1 18:2 18:3 20:1 22:1
Lard 1.5 24.8 3.5 0.5 13.6 45.0 9.9 0.5 0.7
Corn oil
Soybean oil
Oro rapeseed oil
Regular rapeseed oil
10.6
9.9
4.6
3.4
1.5 25.1 60.5 1.9 0.5
4.0 25.7 52.1 7.6 0.7
1.6 58.6 22.6 8.5 2.4 1.9
1.2 21.2 17.4 7.5 13.3 36.1
Mixed rations #3 #1 #2 1.7 0.4 0.3 20.6 10.4 5.9 3.0 0.5 1.7 9.5 4.0 36.7 25.1 54.1 24.4 50.4 24.7 1.7 7.3 8.0 1.1 0.9 2.4 1.6
#4 0.2 4.2 0.5
1.4 21.0 19.8 7.4 12.0 32.2
aExpressed as percent of methyl esters sections of selected tissues were stained with Oil Red 0 using Lillie and Ashburn's method (7).
STATISTICAL EVALUATION Body weights, feed conversions, relative weights of organs and blood values were subjected to analysis of variance and the means compared by Duncan's multiple range test (10).
RESULTS BODY WEIGHTS AND FEED CONVERSION
TABLE III. Average Body Weight (g) and Feed Conversion (g feed/g body weight gained) of Chickens, Ducks and Turkeys Fed Control, 25% Soybean Oil (SBO), 25% Oro Rapeseed Oil (ORSO) and 25% Regular Rapeseed Oil (RRSO) Rations
Body weight at 28
at 49
0-49
days
days
days
724.3a
1522.48
1.47a
667.5a
1430.98
1.74b
467.7b
1058.4b
1.67b
Ducks Control 25% SBO 25% ORSO.
1655.5a
2871.7a
1532.5a
2715.08
1.88a
1582.58
2831.3a
2.29b
275.5b
-c
c
Turkeys Control .... 257o SBO .. 25%o ORSO. 25% RRSO.
819.1a 758.4a 746.3a
2383.2a 2241.98 2035.7a
1.318 1.36a
Chickens Control
25% SBO 25%o ORSO. 25% RRSO.
25%o RRSO.
No significant differences were noted among the average body weights of the birds of the same species fed the control, SBO or ORSO rations throughout the experiment. The average body weights of all species fed the RRSO ration were significantly less (P
