Effect of vitamins K, D, and C on calcium and phosphorus in goat's milk. II. On the effect of vitamin D injection-香川大学学術情報リポジトリ

Vol. 15, No. 2 (1964)

EFFECT O F VITAMINS

K, D,

AND

C

ON CALCIUM

AND PHOSPHORUS IN GOAT'S MILK

II

On the Effect of Vitamin D Injection Toyoki MIYABE

In the previous paper") i t was reported that vltamin K , when injected to dairy goat, caus- ed an increase of the diffusible calcium or phosphorus together with a decrease of produc- tion of calcium phosphocaseinate. I t is well-known that the entire effect of vitamin D is to incxease the availability and retention of calcium and phosphorus and their utilization for proper mineralization of skeleton. Except in a few particulars, an exact mechanism in which these effects are produced has not yet been known. Little is also known about t h e action of vitamin D affecting the synthetic process of calcium phosphocaseinate in mammar y gland.

This experiment was set up to obtain the results a s described in the later pages on goats that were injected vitamin D for each of middle lactation period.

Materials and Methods

Three Saanen dairy goats of middle lactation period were used in this study. And 300,000 I . U. of vitamin D was injected into the intramuscular portion of buttock of goat. This vitamin D ( Eisai Co. ) contains 100,000 I . U. ( calciferol

2

5 mg in 0.5 ml dissolved i t in oil. Sampling of milk was done twice at

8

or 24 hours after injection early in the morning These goats had been fed exactly on the same ration and milked in the same way thrice a day a t the same interval during about 30 days before the present experiment.

As noted in the previous paper(') calcium was determined by Yanagisawa rnethodc2) and phosphorus by Fiske Subbarow method@) using T8ky6 Kaden photometric calorimeter.

The procedures of diffusible calcium and phosphorus determination weIe also carried out as described previously. T h e divided quantitative analysis of protein in milk was made by Rowland methodc4). The protein was determined by Kjeldahl analysis for nitrogen, and the factor used for milk protein was 6-58.

Results

The milk yields secreted from each goat ( I

,

Il

, and

III)

were 4.5, 2.4, and 2.0 L , re- spectively. Their lactation periods were 4.5, 4, and

4

months and number of times of kid parturition were 3 , 1, a n d

2,

respectively.

The injection of vitamin D, a s shown in Table

I ,

caused no consistent changes in titrata- ble acidity and alcoholic reaction. But if observed in detail, it was accompanied by a slight decrease in acidity, as found at 8 hours of goat

IU.

And the alcoholic reactions exhibited already a strongly positive degree at the start before injection.

Table 1. Effect of injection of vitamin D on titratable acidity and alcohol test of milk.

--

No. Goat I

Amount of vitamin D

1 -

3x105 I.

u.

This reaction also was found to be reduced slightly at 8 hours of goat I

.

In the later page present the following tables ( 2 to 4 ) of the subject : the effect of vita- min D on protein, calcium, and phosphorus, and on calcium or phosphorus-combining ca- pacity against casein when injected to dairy goats.

Due to the intramuscular injection of vitamin D, not so evident change was recognized in the phosphorus or diffusible phosphorus contents, but the protein

,

casein, calcium, and diffusible calcium contents all exhibited clearly the same tendency to decline with lapse of time. And its effect was so smaller in the protein content than in the calcium content.

- - .- - --

Hours after injection 1 0

/

8 1 2 4 1 0 1 8 1 2 4 1 0 1 8 I 2 4 0 8 1 2 4

Table 2 Effect of injection of vitamin D on protein, calcium, and phosphorus in milk

-- -.

No. Goat I Goat II. Goat

IK

- 1

3x106 I. U.

Aver age Amount of vitamin D 3x106 I . U. 3x106 I. U.

Goat II. Goat

rn

-1

3x106 I. U. 3x106 I. U.

Titratable acidity 70% alcohol test

Hours after injection

Average 19.5 21.2 4.l Total protein g/dl Casein g/dl

Casein in 100 of total protei~ Total Ca mg/dl Diff. Ca mg/dl Diff.Ca in 100 of total Ca - Total P mg/dl Diff. P mg/dl Diff. P in 100 of total P

Table 3. Effect of injection of vitamin D on calcium and phosphorus combined with casein. 20 5 4.t

--

21 5 Stt 14.0 # -

-No, Goat I Goat II. Goat l!I

-

Amount of vitamin D 3x106 I. U. 3 X 10s I. U. Average

- Hours after injection

17 7 14.0

ttt

15.0 ttt 13.8 # .- 17.5 ttt 18 9 - -25.5 9.3 3.07 Bound-Cajmg)pescasein(g) Bound-P(mg)pescasein(g) Bound-Ca/P 15.8 i t i # - k , # i t t 15.6 11.9 1.31 2 5 7 12.2 2.11 33.6 12.6 2,67

I

31.2 31.8 19..3 1.95 26.2 131.3 17..6 1.81 15..0 1075 15.7 1.99

Vol .. 15, No. 2 (1964) 157 T h i s was not true for every individual of goats, but the majority showed this tendency. Although the injection of vitamin D had an effect on the variation of the protein or casein content, the magnitude of the effect differed among the individuals ; it was much more remarkable in goat I than in goats

II

and IIt. The rate of change in the protein of vita- min D seems to be similar to that in the protein of vitamin K.

The content of protein in milk showed a decrease of a quite similar rate a s compared with that of casein in milk. Thus, since there was no difference between t h e two, the r a t e of casein in 100 of total protein ( i . e . , casein distribution) did not seem to have a definite variation likewise. That is to say, the decrease of protein content may also mean

Table 4.. The average rates of increase and decrease after injection of vitamin D.

Amount of vitamin D Hours after injection Total protein (%)

Casein (%)

Casein in 100 of total p~otein (%)

that of casein content. At the same time, much more ema ark able decrease occurred in the calcium and diffusible calcium contents, particularly in the latter content.

I t will be noted that the variation of calcium content occurred remarkably, a s the result of injection of vitamin D, without change of phosphorus content, where the variation was rather larger in the diffusible calcium content than in the calium content.

I t is well-recognized that vitamin D stimulates the absorption of ion-calcium from in- testinal track, but does not appear to exert a direct effect on the absorption of phosphate. I n addition, vitamin D appears to function in the internal tissues. In this experiment, i t seemed to suggest t h a t vitamin D interfered with the secretion of calcium i n mammary gland, so that less calcium was secreted. The injection of vitamin D i s of little or of no important insofar as phosphorus content in milk is concerned, a s compared with calcium content in milk. Then the ratio of Ca/P showed a tendency to decline by about 16 per cent as total average value and no major difference was observed as compared with the case of injection of vitamin K.

Without regard to the decrease of calcium content, the calcium was recognized to be combined with casein by about twice a s compared with the value before injection, a s

Goat I -- 3x106 1.U Ave

.

8

1

24 Total Ca (%) Diff

.

Ca (%) Diff. Ca in 100 of total Ca (%) - - Total P (%) Diff. P (%) Diff. P in 100 of total P (%) Goat 11 -26 7 0.0 Goat JE - Ca / P

/

-8.6 1-6.5

/

-7.6 1-15.6 1-18 0 1-16.8 -15 2 -5.7 -37.1 -63.2 2.4 -3 5 -5.7 -26.6-249-25.8 -32 5 -10 0 -29.6 -5.0 Total 3x106 I.U. 3x1051 U. Ave. average 8

1

24 5 8 -13.1 -17.9 - 13.4 18.3 4 2 -2.1 4.1 6 3 - -3.1 -10 0 -7.2 -16 2 -16.7 -17.2 2 5 -5 5 -32.3 22.5 14 2 -18 3 2 4 -0.1 -2.4 7.6 6 2 -8.6 5 2 6.3 -6.3 -- 0.1 -25.1 -25.6 7.9 7.4 -0 8 -17.2 -13.4 5.9 -2.5 4 8 7.4

158 Tech.. Bull,, Fac Agr. Kagawa Univ.. found in goat I An instance was found where considerable depression in the bound-calci- um occurring a t the rate of about 20 per cent, a s shown in goat

II.

The bound-phos- phorus displayed the same tendency a s the bound-calcium, although to a lesser degree and considerably more irregularly. This phase was contrarily indicated a s compar ed with vitamin K. The alcoholic reaction was changed into positive from negative by the injection of vitamin D. This may be considered t o be due to the fact that vitamin D has a function to combine calcium with casein in milk and the ratio of bound-Ca/P becomes evidently high and unbalanced.

Summary

An experiment was carried out in which vitamin D was injected to three Saanen dairy goats for the manner of calcium and phosphorus to calcium phosphocaseinate in milk which was secreted in mammary.

And 300,000 I . U . of vitamin D was injected into the intramuscular portion of buttock of goat ..

The present data, as the results of injection of vitamin D , indicated that much more I e - markable decrease occurred in the caIcium and diffusible calcium contents in milk, Par- ticularly in the latter content, without change of the phosphorus and diffusible contents in milk. From these facts, it seemed to suggest that vitamin D interfered with the se-, cretion of calcium in mammary gland, so that less calcium was secreted.

And vitamin D did not appear to exert a direct effect on the secretion of phosphorus. Nevertheless, vitamin D had a n action of binding the calcium to casein regardless of r e - markable decrease of the calcium content in milk And the tendency exhibited to decline the casein production.

(Received Decembe~ 10, 1963)

References

(I) MIYABE, T . : Tech. B u l l , , F a c , A g r . , K a g a w a (3) SUBBAROW, F . S.. :

,I

Biol.. C h e m . , 66, 375

Univ.,lS, 149 (1964). (1925).

(2) YANAGISAWA, F. : Seikagaku Zikkenhi3, 1, (4) R O W L A N D , S. J , : l . D a i r , y Res.. , 9 , 42 (1938)..

Vol. 15, No. 2 (1964) 159

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