The Vitamin A and Vitamin D Content of Cod Liver Meal*

The Vitamin A and Vitamin D Content of Cod Liver Meal*

THE VITAMIN A AND VITAMIN D CONTENT OF COD LIVER MEAL* ETHEL M. CRUICKSHANK.t Ε Β. HART AND J. G. HALPIN (From the Departments of Agricultural Chemist...

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THE VITAMIN A AND VITAMIN D CONTENT OF COD LIVER MEAL* ETHEL M. CRUICKSHANK.t Ε Β. HART AND J. G. HALPIN (From the Departments of Agricultural Chemistry and Poultry Husbandry, University of Wisconsin, Madison)

VITAMIN D

In testing the vitamin D content, nine groups of chicks (one day old Leghorns ) were used ; seven birds in six of the groups and six in the remaining three. These birds were housed in pens in an attic where direct light had no access. They were kept on screens in place of shavings. The basal ration used was one that will invariably lead to a rachitic condition in chicks in 5 to 6 weeks in the absence of the antirachitic vitamin or its equivalent, ultra violet light. The constituents were 97 parts yellow corn. 2 parts CaCOg, 1 part NaCl. Skimmed milk in addition was allowed ad libitum. The rations of the different groups were as follows : Group I Basal ration. Group II Basal ration + irradiation (10 minutes daily except Sundays). Group I I I Basal ration -\- 2% cod liver oil. Group IV Basal ration-|- 1 % cod liver meal. Group V Basal ration + 3% cod liver meal. Group VI Basal ration + 5% cod liver meal. Group VII Basal ration -)- 7% cod liver meal. Group VIII Basal ration + 10% cod liver meal. Group I X Basal ration + 2% cod liver meal -\- 2% Casein. * Published with the permission of the Director of the Wisconsin Agricultural Experimient Station. IFellow of British Ministry of Agriculture and Fisheries. 9

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(Received for Publication 7-20-27) Since cod liver meal is now coming on the market in con­ siderable quantities as an ingredient for poultry feeds it was believed important to ascertain the vitamin A and vitamin D potency of this material by testing it with chickens. The particular meal used was obtained in 1926 from St. John's Newfoundland and had a high oil content, viz., 45 per cent ; while it also contained 43 per cent of protein.

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VITAMIN A

To test the vitamin A content of the meal, the experiment was repeated, save that in this case white "corn (which lacks vitamin A) was used in place of the yellow. In this experiment (No. I I ) , however, the cod liver oil used for the control group proved

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Casein was added in the ease of Group I X in order to raise the protein level, as it was thought that possibly part of the beneficial action of the cod liver meal might be attributed to the high protein content. These rations thus provided a group where no vitamin D was supplied, a group where its place was taken by ultra violet light and one where it was supplied in the form of cod liver oil ; while the other groups were designed to show the efficiency of various levels of the cod liver meal in preventing the development of rickets. The birds were kept on these rations for 6 weeks, by which time Group I was exhibiting symptoms of rickets, viz., inactivity, awkwardness of gait and ruffled condition of feathers. The extent of calcification was measured by the silver nitrate test and also by the ash analyses of the alcohol extracted tibias. In carrying out the silver nitrate test the tibia was carefully dissected out, freed from adhering tissue and placed in 10 percent formalin. At the end of three days the bone was removed and washed free from formalin and a longitudinal section cut from the proximal end. This section was treated with 1.5 per cent AgNOg and exposed to the light so that the calcified area becomes black while the cartilage retained its yellowish white color, and an indication of the extent of calcification was thus obtained. The following table shows the ash analyses of the bones from the different groups. From these figures it will be seen that the chicks on the basal ration only, were markedly rachitic, the ash of the tibia varying from 33 to 37 per cent; while in the case of groups II and I I I where the antirachitic factor was abundant, the ash ranged froiii 46 to 52 per cent. The groups getting difi:erent levels of cod liver meal showed a normal ash even when getting only 1 per cent of cod liver meal. The silver nitrate test confirmed these findings, showing good calcification in all the groups save No. I where there was a wide uncalcified area of proliferating cartilage.

VITAMINS A S H ANALYSES or

IN COD LITER

MEAL

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T I B I A S OF S I X - W E E K S OLD C H I O K S GETTING COD L I V E B MEAL AS A SOURCE OP V I T A M I N D

Group I Basal R a t i o n

Basal -f I r r a d i a t i o n

Per-cent of asli

Number of bird

Per-cent of a s h

Died 35.37 Died 37.15 34.35 33.89 37.05

6442 6451 6460 6469 6478 6487 6496

50.95 52.46 45.73 47.84 46.73 Died 50.63

Group V Group IV Basal + 1% Cod Liver Basal + 3 % Ood Liver Meal Mea I 6444 6453 6462 6471 6480 6489 6498

45.10 49.18 46.36 Bone lost Died 49.94 48.39

Group VII Basal + 7% Cod Liver Meal 6447 6456 6465 6474 6483 6492

47.92 49.59 45.20 Died 43.82 45.71

6445 6454 6463 6472 6481 6490 6499

45.35 47.05 44.95 44.89 44.13 47.82 44.51

Group V I I I Basal + 10% Cod Liver M«al 6448 47.00 6457 46.95 47.63 6466 6475 43.70 6484 47.41 48.12 6493

Group I I I Basal + 2 % Cod Liver Oil Number P e r cent of a s b of bird 6443 6452 6461 6470 6479 6488 6497

48.88 47.13 Died 47.26 46.87 Died 49.67

Group VI Basal + 5% Cod Liver Meal 6446 6455 6464 6473 6482 6491 6500

Died 46.56 46.95 44.32 44.95 47.17 45.71

Group I X Basal -)- 2 % Cod Liver Meal + 2.5% Casein 6449 6458 6467 6467 6476 6485 6494

48.00 47.85 42.85 42.85 46.00 46.90 46.21

to have little or no vitamin A in it, so a third experiment (No. I l l ) was run, using a different brand of cod liver oil. The basal ration used was 92 parts white corn, 5 parts yeast, 2 parts CaCOg and 1 part NaCl, plus skimmed milk ad libitum. Yeast was added to insure an abundance of vitamin Β as some of the symptoms exhibited by the birds in experiment I I closely re­ sembled those of polyneuritis.

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Number of bird 6441 6450 6459 6468 6477 6486 6495

Group I I

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Among these groups, therefore, No. I had neither A nor D supplied; No. II had D supplied by irradiation; No. I l l was getting both A and D from the cod liver oil; while the remaining groups from 4 to 8 inclusive had sufficient D in the ration from the cod liver meal, and it could be determined whether they were getting enough A from the different levels. Groups IX and X had irradiation in addition to the cod liver meal to ensure that an insulBciency of vitamin D was not complicating the results. Group I came down early—in about 4 to 5 weeks. The symptoms exhibited were staggering gait and loss of control, accompanied by emaciation and weakness and a general ruffled condition of the feathers. Ophthalmia, however, was not apparent. Post-mortems were done on seven birds and of these two had pneiumonia; 3 had nodular lesions in mouth and pharynx; while all showed kidney lesions. In these lesions the kidneys were pale in color or mottled grey, due to the fact that the renal tubules were filled with white material, which proved to be urates. The ureters also were frequently distended and filled with these urates. In two cases a urate deposit was found in the pericardial sac and on the liver as well as in the kidneys. In the second group where only vitamin A was deficient the same staggering movement and lack of control was observed. Both in this group and Group I trembling was noted in several of the chicks. Autopsies were performed on 5 and it was found

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In the third experiment there were 10 groups of ten chicks each (Leghorns and started when one day old), and the rations fed were as follows : Group I Basal ration. Group II Basal ration + irradiation (10 minutes daily). Group I I I Basal ration + 2% cod liver oil. Group IV Basal ration -f- 1% cod liver meal. Group V Basal ration + 3% cod liver meal. Group VI Basal ration + 5% cod liver meal. Group VII Basal ration -|- 7% cod liver meal. Group VIII Basal ration -|- 10% cod liver meal. Group IX Basal ration + 3 % cod liver meal + irradiation. Group X Basal ration + 5% cod liver meal + irradiation.

VITAMINS

IN COD LIVER

MEAL

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that only one had nodular lesions in the mouth and pharjnx ; submaxillary edema ivas found in two, Avhile kidney lesions were present in all. In Group 111 (cod liver oil group) the chicks appeared to be in excellent condition. Their appetite was always good and

weight increase was satisfactory. On autopsy all organs were found to be normal. One marked difference between this group and the A-deficient lots was the deposition of fat in the abdominal cavity of the normal birds. Group IV which were receiving 1 per cent cod liver meal

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FiGUBE I

Effect of Vitamin A Deficiency in Chicks 6 to 7 Weeks Old. Left— A normal chick which received 2 per cent of cod liver oil plus the basal ration. In this specimen t h e r e were no kidney lesions or u r a t e s in t h e ureters. Right—A chick t h a t received 5 per cent of cod liver meal plus the basal ration. Note the mottled kidneys and u r e t e r s distended with accumulated u r a t e s .

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were in very poor condition. Two of the birds showed typical ophthalmia. Autopsies were performed on eight of them and of these six had nodular lesions in the mouth and pharynx and in one case these extended down the oesophagus. Five had more or less marked submaxillary edema, while kidney lesions were present in each bird examined. Group V manifested the symptoms of vitamin A deficiency in practically the same degree as the preceding group. Out of 8 autopsies three showed nodular lesions and three submaxillary edema. In some birds the eyes were swollen, and there was some watery discharge, but the condition was not that of typical ophthalmia. Lung lesions were found in two cases, while 7 showed kidney lesions and marked distension of the ureters with urates. Group VI. Evidently the A requirements were not supplied even by 5 per cent of cod liver meal though the weight increase here was better than in lots IV and V. Nine autopsies were made. Five had nodular lesions in mouth and pharynx, six had submaxillary edema, one showed congestion of the lungs, and kidney lesions were found in eight. Two of these birds had been killed and the kidney lesions in these were very slight. Group VII. Five birds were alive in this lot at the end of the experiment as compared with 2 in Group VI and none in Group V. Out of 10 autopsies 6 had lesions in mouith and pharynx, 8 had submaxillary edema (6 of these being slight) and one had typical ophthalmia. One had pneumonia and 7 had kidney lesions. Group VIII. (10 per cent cod liver meal) was the best of all the groups, save the control, though it was not normal. Eventually the movement of the chicks became rather bad, but the cod liver meal at this level had enough vitamin A to delay to some extent the onset of the symptoms. Post-mortem examination of 8 birds showed that 4 had slight submaxillary swelling, and there were some slight lesions in mouth and pharynx, but these were not marked. Five had kidney lesions, but the majority of these were slight. Nutrition was markedly better than in the other groups, with the exception of the control. Group I X which had 3 per cent cod liver meal plus irradiation

VITAMINS IN COD LIVER MEAL

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DISCUSSION

From the foregoing data it will be seen that this particular sample of cod liver meal contained enough vitamin D to protect the chickens against rickets even when fed at levels of 1 per cent and 3 per cent. Since the meal contained 45 per cent of oil, they were only slightly less than 5 per cent cod liver oil in their ration when 1 per cent of the meal was fed, which Hart, Steenbock and Lepkovsky^ pound sufficient to promote good calcification in chickens. With regard to the vitamin A content of the meal, even 10 per cent was not sufficient to protect the birds, though it was evident that some vitamin A was present, since the group receiving 10 per cent of meal continued to put on weight, survived longer, and did not show pathological symptoms in such a marked degree as the other groups. The clinical symptoms which were most obvious were the staggering gait and loss of control which were particularly marked in these lots getting the lower levels of meal. In many cases and for some time before the birds died spasms of trembling were observed. Occasionally the neck was twisted to one side so that it was almost impossible for the birds to eat. This also happened when the loss of control became very marked, so that lack of food probably ultimately hastened death in many instances. Nodular lesions in the mouth and pharynx, though not found in every ease, were very prevalent, and, according to Beaeh^ are not associated with any other disease or deficiency. Frequently there were small lumps of caseous matter in the pharynx, and sometimes they blocked the larynx and probably were the primary cause of death.

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was no better than the corresponding Group V which was not irradiated. Out of 8 autopsies 4 showed submaxillary edema, 4 had lesions of mouth and pharynx, and one showed ophthalmia. Pour showed kidney lesions and in two of the birds there was a slight deposition of urates in the pericardial sac. Group X gave results very similar to the preceding group. Six of the birds autopsied had submaxillary edema and nodular lesions of the mouth and pharynx, one had ophthalmia of both eyes and one had pneumonia. Four had kidney lesions.

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SUMMARY

The sample of cod liver meal used in this experiment contained sufficient vitamin D to promote good calcification in chickens when fed at levels of 1 per cent and 3 per cent. Even 10 per cent of the cod liver meal was not sufficient to protect chickens against a vitamin A deficiency. BIBLIOGRAPHY 1 HART, E . B . , STEENBOCK, H . AND LÎEPKOVSKÏ·, S., J. Biol. Chem. 1925,

Ixv, 571. 2 BEACH, J. R., Am. Vet. Med. Assn. 1921, xi, 303.

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Submaxillary edema was a frequent symptom, and there seemed to be somia tendency to edema in the body itself. Kidney lesions were found in practically all the birds that died during the experiment. In the birds that were killed at the termination of the experiment these lesions wre found to be very slight, though judging by external symptoms many of the birds were in a very weak condition and unlikely to live more than a day or so longer. I t seems probable therefore that the accumulation of urates in the ureters occurs only shortly before death. Though all the birds that died from vitamin A deficiency in this experiment showed kidney lesions, a finding that is in accordance with the work of Beach, Hinshaw and others, this condition is apparently not confined to this deficiency. In some field cases that were brought to our notice the same lesions were found in eoccidiosis, where the crops Avere full of yellow com, indicating that the lesions were due to the eoccidiosis rather than to an A deficiency.