V itam in D Studies and Rickets /
-By
H. ERNEST BECHTEL
<1
A THESIS
1 ’resen ted to the G raduate School of M ichigan State C ollege of Agriculture
and A p p lied Science in Partial F ulfillm ent of Requirements
for the D egree o f D octor o f Philosophy
Chem istry Departm ent
East Lansing, M ichigan
1935
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R eprinted from T h e J o u r n a l o p N u t r i t i o n
Vol. 11, No. 6 and Supplement, June, 1936
A STUDY OF THE SEASONAL VARIATION OF
VITAMIN D IN NORMAL COW’S M IL K 1
H. ERNEST BECHTEL AND C. A. HOPPERT
Departm ent of Chemistry, Michigan State College, E ast Lansing, Michigan
T H R E E FIGU RES
(Received for publication February 7, 1936)
In recent years a great deal of attention has been given
to the importance of vitamin D in the human dietary and
particularly to the desirability and means of enhancing the
milk supply in this factor. The methods by which the latter
has been achieved are well known and relatively little thought
is now given to the antirachitic potency of milk produced
under ordinary farming conditions. Although many studies
have been made of the vitamin D content of cow’s milk, none
of them has been of a comprehensive nature. Table 1 sum
marizes the results of some of these investigations.
Similar trends in variations of the vitamin D potency of
milk are indicated by the work of other investigators. Luce
( ’24) found milk from pastured cows definitely richer in vita
min D than that from stall-fed cows kept in the dark. Luce
concluded that the concentration of the antirachitic factor in
milk depended on the ration and possibly also on the degree
of illumination received by the cow. Chick and Roscoe ( ’26)
found 0.25 gm. of milk fat from pastured cows equal to 0.60
gm. milk fat from cows fed green feed in a dark stall. Ex
posure of the cow to outdoor light without a change in ration
resulted in a twofold increase in the vitamin D content of
1 The data in this paper -were taken from the thesis presented by H. Ernest
Beehtel to the faculty of the Graduate School of Michigan State College in 1935
in partial fulfillment of the requirements for the degree o f doctor of philosophy.
537
T H E JO U R N A L, OP N U T R IT IO N , VOL. 1 1 , N O . 6 AN D S U P P L E M E N T
538
H . E. BECHTEL AND
C. A . H O P P E B T
the milk fat. Dutcher and Honeywell ( ’27) examined some
Kansas butter samples and found that milk fa t from cows
exposed to sunshine was superior in vitamin D potency to fat
produced by cows fed in the dark.
These and other studies which might be mentioned indicate
in a general way the variations that occur in the vitamin D
potency of milk and the factors which contribute to this varia
bility. However, most of the work done has been of a rather
fragmentary nature so that it seemed desirable to make a
more extended study of the subject.
TABLE 1
V I T A M I N D C O N T E N T O F N O R M A L M IL K
U N IT S P E R QUART
SOURCE O F DATA
U.S.P.*
Steenbock
21.3
36.5
7.9
13.5
Mitchell and associates (Pennsylvania) ( ’32) Summer milk
16.2
6.0
Krauss (Ohio) ('33)
Winter milk
Summer milk
7.6
16.2
2.8
6.0
Russell (New Jersey) ( ’33)
Summer milk
Always less than
43.2
16.0
Steenbock and associates (Wisconsin) ( ’30)
June 1925
September 1925
1 The values in this column were obtained by multiplying the number of
Steenbock units by 2.7.
MATERIALS A N D METHODS
This study was begun in 1932, the milk being derived from
various sources. Milk from the Holstein and Guernsey herds
at Michigan State College was assayed monthly over a period
of 2 years. Only the higher producers, namely, cows that
were being milked three times daily were used, the average
number of Holstein and Guernsey cows being fourteen and
eight, respectively, for the 2-year period. From these cows
100-pound portions of composite 24-hour samples were saved
monthly.
Monthly samples of a similar nature were derived from a
few of the highest producing cows in the Holstein herd of the
Michigan Experiment Station. These cows were kept on a
ration of alfalfa hay, silage and corn.
VA K IA TIO N OF V IT A M IN D I N
M IL K
539
From July 1933 to July 1934 monthly samples of Michigan
State College Creamery butter, made from milk produced by
local Michigan dairymen, were also put aside for vitamin D
assays. Approximately fifteen herds, consisting largely of
grade Holstein cows were represented in this group of
samples.
In the case of the milk a small portion was used for a fat
determination, the remainder being run through a cream
separator. The butter obtained by churning the cream was
packed in paper cartons and stored at 0°F. until the samples
were to be assayed. At this time the butter was heated on a
steam bath for about an hour and the relatively pure milk
fat upon which the assays were conducted was suctioned off.
Except when needed for assay, the fats were kept at 0°F.
The vitamin D determinations were carried out by the
curative feeding technic, several changes being made in the
official procedure. The Steenbock basal ration was slightly
modified to obtain more consistent and somewhat better
growth of the rats during the preliminary period. The
rachitogenic diet used throughout this study was composed
of the following:
P e r cen t
Yellow cornmeal
38.0
Oatmeal
37.5
Wheat gluten
20.0
Calcium carbonate
3.0
Sodium chloride
1.0
Yeast powder
0.5
Instead of feeding the fat as a daily supplement during
the first 8 days of the test period, the entire amount was mixed
with 40 gm. of the basal diet. This mixture was found to be
consumed in 6 to 8 days after which the basal diet was fed to
finish the 10-day period. Control rats receiving 29 mg. of
Official Reference Oil equivalent to 2.7 U.S.P. units were used
for comparison. The usual staining technic was applied to
the radii and ulnae.
In carrying out the assays a preliminary test was made
to determine the approximate vitamin D content of the vari
540
H . E. BECHTEL AND
C. A . H O P P E R T
ous samples. The confirmatory tests were then conducted at
three levels using three to five rats at each level. ^
It became apparent early in this investigation that fats
of low potency could not be assayed because of the limited
capacity of the rats to consume fat. Amounts up to 6 and
8 gm. were consumed fairly consistently. However, when the
dosage was increased to 10 gm. approximately only half of the
test animals consumed the fat-basal diet mixture in 8 days.
Attempts were therefore made to effect a concentration of the
vitamin D so that fats of lower potency might be assayed.
Although the work of Kon and Booth ( ’34) indicated that at
least a part of the vitamin I) of butter fat was unstable and
could not be recovered quantitatively in the non-saponifiable
matter, their method as well as several modifications were
given a trial. Sometimes the recovery of vitam in D was
quantitative, but more often it was not, so that this method
of concentration was abandoned.
Inasmuch as the concentration of vitamin D in cod liver
oil may be accomplished by extraction of the oil with alcohol,
this method was next tried and proved to be satisfactory. The
fats of low potency were therefore treated in the following
manner: 100 gm. of melted milk fat was placed in a separa
tory funnel previously warmed in a 37 °C. oven and 100 cc.
of hot ethyl alcohol (95 per cent) was added. The mixture
was then shaken fairly vigorously and the funnel placed into
the 37° C. oven until the layers had separated. The fa t was
then drawn off into a warm beaker and the alcohol layer into
a 500 cc. volumetric flask. The fat was returned to the sepa
ratory funnel and the beaker rinsed with 50 cc. of hot alcohol
which was then added to the fat. The mixture was again
shaken and the layers allowed to separate in the oven. The
separations were made as before and three additional ex
tractions carried out with 50 cc. portions of hot alcohol. B y
this process approximately 20 per cent of the fat was removed
and this fat contained all of the vitamin D. The combined
extracts were brought to a volume of 500 cc. with ether in
order that the fat would be kept in solution. It was observed
V A R IA T IO N OF V IT A M IN D I N
541
M IL K
that the antirachitic value was retained for at least 2 months
when the extract was stored at 0°C. Aliquots of this solution
were poured on 40 gm. portions of the rachitogenic diet in
evaporating dishes and the ether and alcohol allowed to
evaporate spontaneously. These mixtures were then fed in
the usual manner.
TA BLE 2
Antirachitic potency of milk f a t obtained from the Holstein h erd1
DATE
F A T -C O N T A IN
IN G 1 U .S .P .
V IT A M IN D
U N IT
AVERAGE D AILY
PR O D U C T IO N P E R COW
FAT IN
M IL K
V IT A M IN D P E R
QUART O F M IL K
Fat
V itam in D
U.S.P.
gm.
gm.
U.S.P.
per cent
1.5
1.1
1.1
3.3
817
717
862
748
545
701
784
227
3.26
2.88
3.53
2.78
—
—
—
—
—
—
—
—
—
—
—
—
January
February
March
April
May
June
5.6
9.3
3.7
3.7
3.0
826
875
835
748
730
148
94
226
202
243
3.15
3.26
4.6
3.1
6.7
5.9
7.0
1.7
1.1
2.5
2.2
2.6
—
—
—
—
—
—
July
August
September
October
November
December
1.5
1.9
1.5
3.7
7.4
776
862
780
690
753
517
454
520
186
102
3.20
3.28
2.98
2.96
3.19
16.6
14.9
15.5
5.5
3.2
6.1
5.5
5.7
2.0
1.2
—
—
—
—
—
—
—
7.4
7.4
4.4
3.0
—
844
939
871
898
—
114
127
198
299
—
3.17
3.29
3.06
3.31
—
3.6
4.2
6.1
9.9
—
St.2
AVERAGE
D A IL Y
A M O U N T OF
S U N S H IN E
hours 3
1932
July
August
September
October
November
December
17.8
20.2
27.7
6.3
6.6
7.5
10.3
2.3
11.5
10.0
7.8
3.3
4.0
3.0
1933
2.97
2.93
2.89
4.1
7.1
4.4
6.8
8.5
12.7
12.8
10.8
7.1
6.0
2.0
1.4
1934
January
February
March
April
May
1.3
1.6
2.3
3.7
2.3
5.9
5.0
6.0
11.3
1 Average of fourteen cows per month.
2 Steenbock.
’ Average amount of available sunshine according to the East Lansing Weather
Bureau.
H . E. BECHTEL AND
542
C. A . H O P P E R T
The above method permitted practically complete recovery
of the vitamin D from samples of milk fat of which 2 to 10 gm.
had to be fed of the original fat to get the typical narrow
continuous line of calcification. Inasmuch as there was no
way of checking the fats of lower potency directly, it had to
be assumed that the method was also satisfactory for such
TABLE 3
Anti/raohitic potency of milk fa t obtained from the Guernsey h e rd 1
f a t -c o n t a i n
DATE
1 U .S .P .
V IT A M IN D
U N IT
in g
AVERAGE D A IL Y
PR O D U C T IO N P E R COW
FAT IN
M IL K
V IT A M IN D P E R
Q UART O F M IL K
U .S.P.
St.2
AVERAGE
D A IL Y
A M OUNT OF
S U N S H IN E
Pat
Vitamin D
gm.
gm.
U.S.P.
1.1
1.1
1.3
3.0
3.7
3.7
649
685
708
699
708
930
590
623
545
233
191
251
4.93
4 .86
5.15
4 .52
' 4 .02
4.76
4 3 .8
43.1
38 .7
14.7
10.6
1 2.6
1 6.2
16 .0
1 4.3
5.4
3.9
January
February
March
April
May
June
4 .4
9.3
3.7
3.0
3.0
907
835
762
658
694
206
90
206
219
231
4.42
4 .57
4.33
4.34
4.42
9.8
4.8
11.4
14.1
14.4
3.6
1.8
4 .2
5.2
5.3
—
—
—
—
—
—
July
August
September
October
November
December
1.5
1,9
1.9
2.6
3.7
—
744
853
694
721
912
—
496
449
365
277
246
4.43
5.09
4.61
4.76
4.86
28.8
26.2
23.7
17.9
12.8
1 0.7
9.7
—
—
—
—
1 2.8
1 0.8
7.1
6.0
2.0
1.4
2.2
2.9
4 .4
5.9
2.3
5.9
5.0
6.0
11.3
per cent
hours 3
1932
July
August
September
October
November
December
4 .7
1 1.5
10.0
7.8
3.3
4 .0
3.0
00
00
1933
6.6
4 .7
4.1
7.1
4 .4
6.8
8.5
1 2.7
1934
January
—
February
7.4
March
5.6
April
3.7
May
3.0
1 * ____
1 Average of eight cows
1 Steenbock.
—
—
_
866
835
826
871
117
149
223
290
4.44
4.50
4.52
4.90
5.9
7.8
11.9
15.9
per month.
3 Average amount of available sunshine according to the E ast Lansing Weather
T A BLE 4
Antirachitic potency of milJc fa t obtained from the experiment station h erd 1
FAT C O N T A i:
1 TJ.S.P. V II
D U N IT
lb.
gm .
gm .
U.S.P.
p e r ce n t
24.2
26.3
26.4
23.1
26.0
24.9
—
—
5.5
25.9
20.9
24.0
15.1
13.8
8.3
4.4
3.5
4.1
1.3
1.5
1.5
2.6
3.7
3.7
544
617
581
508
662
435
418
411
387
195
179
118
2.30
2.64
3.18
3.34
4.02
3.29
ZM
MJ
U .S .P .
CS
&
E*
55
DAILY AM OU
S U N S H IN E
Corn
lb.
V IT A M IN D
P E R QUART
M IL K
Vitamin D
Silicon
|
Alfalfa
Ph
lb.
DATE
1933
July
August
September
October
November
December
AVERAGE D A IL Y
PK O DUO TIO N
PER, OOW
o£
£a
AVERAGE D A IL Y
FE E D IN T A K E
m
h o u rs 3
17.3
17.2
20.7
12.5
10.6
8.7
12.8
10.8
7.1
6.0
2.0
1.4
6.4
6.4
7.7
4.6
3.9
3.2
1934
J anuary
2.3
5.9
February
26.1 26.0
74
3.40
0.3
4.4
7.5
327
2.8
5.0
21.2
121
3.73
March
3.2
4.4
531
8.3
3.1
19.7
2.84
6.0
7.5
2.8
April
23.8
535
13.0
3.7
145
9.3
11.3
13.9
26.0
2.2
794
3.13
5.1
3.6 12.6
361
May
1 Average of five cows per month.
2 Steenbock.
3 Average amount of available sunshine according to the East Lansing Weather
Bureau.
TA BLE 5
Antirachitic potency of creamery milk f a t 1
V IT A M IN D P E R Q UART
OF M IL K
F A T C O N T A IN IN G
1 U .S .P . V IT A M IN
D U N IT
FA T I N
M IL K
gm .
p e r cen t
1933
July
August
September
October
November
December
1.3
1.9
1.3
2.6
4.4
3.0
3.5
3.5
3.5
3.5
3.5
3.5
26.3
18.0
26.3
13.1
7.8
11.4
1934
January
February
March
April
May
June
—
3.7
—
—
3.0
1.9
—
3.5
—
—
3.5
3.5
9.2
—
—
11.4
18.0
DATE
U .S .P .
Steenbock
AVERAGE D A IL Y
A M O U N T OF
S U N S H IN E .
h o u rs 2
9.7
6.7
9.7
4.9
2.9
4.2
12.8
10.8
7.1
6.0
2.0
1.4
—
3.4
2.3
5.9
5.0
6.0
11.3
11.8
—
—
4.2
6.7
1 Average of twelve herds of cows per month.
2Average amount of available sunshine according to the East Lansing Weather
Bureau.
543
CO
00
jteth jo
+* Q)
2•H
CO I
n •3
C Q, O
o
in o
co
w w
(3HTW jo qjtynfo j a i )
sq iu fl a n-pureq-FA M ' S ’Il
m
CO
o>
*qi
St <8 M
h co CM
o>
CO
r—
I 05
r -i
milk from the Holstein and Guernsey herds, and available sunshine.
milk from the Experiment Station herd and M. S. 0. Creamery, and available sunshine.
milk fat from Holstein and Guernsey herds, and available sunshine.
strpqsuns s jnog jo jaqnm.ii ©Sujsav
3
ie>
o
Antirachitic potency of
Antirachitic potency of
Antirachitic potency of
H . E. BECHTEL AND
Fig. 1
Fig. 2
Fig. 3
544
C. A . H O P P E R T
H 05
V A R IA T IO N OF V IT A M IN
D IN
M IL K
545
fats. Quantitative recovery was also obtained when a definite
amount of vitamin D from the official reference oil was added
to milk fat and subjected to hot alcohol extraction.
The results of this study of the seasonal variation in the
vitamin D content of cows’ milk are presented in tables 2, 3,
4 and 5 and portions of the data are shown graphically in
figures 1, 2 and 3. The data include not only the results of
the bioassays but also the average daily production of milk
fat and the number of vitamin D units in the milk fat. The
results were also calculated in terms of U.S.P. units per quart
and these values are presented in figures 1 and 2 with the
average daily hours of sunshine available each month. To
simplify the comparison of these results with those given in
the older literature the antirachitic potency is also expressed
in terms of Steenbock units.
Inasmuch as exposure of the cows to sunlight and the in
gestion of sun cured roughages are two important factors
which influence the vitamin D potency of the milk, a brief
reference to the general management of the several dairy
herds is appropriate. The main Holstein and Guernsey herds
of the college were kept under parallel conditions at all times.
Prom May to September, inclusive, these animals were
pastured an average of 8 hours daily and received no hay
or corn silage. During* October they were pastured an aver
age of 5 hours daily and received about 1 pound of hay per
100 pounds of body weight. Prom November to April, in
clusive, the animals were put out doors in dry lot for an
average of 2 hours daily. During this period in 1932-1933
they received besides their allowance of grain an average of
approximately 2 pounds of hay per 100 pounds of body weight,
no corn silage being included. During the correspondingperiod in 1933-1934 the animals received f pound of hay and
3 pounds of corn silage per 100 pounds of body weight in
addition to grain. The average weight of the Guernsey cows
was 1150 and that of the Holsteins 1400 pounds.
The Holstein cows in the experiment station herd were out
of doors in dry lot an average of 7 hours daily from May
546
H . E. BECHTEL AND
C. A . H 0 P P E R T
to September, inclusive, and about 2 hours daily during the
other months. These cows were kept on a ration of alfalfa
hay, corn silage and corn as shown in table 4.
The general management of the local Michigan dairy herds
which served as the source of the college creamery butter
samples was typical of that practiced in this state. The cows
were fed chiefly home grown feeds consisting largely of alfalfa
and cereal grains and were usually pastured as early and as
late as conditions permitted. They were probably exposed
to sunshine for a longer time than the cows in the college
herds.
Regarding the assaying of the various samples of butter,
there was a considerable interval between the time the
samples were obtained and the time the bioassays were made.
This delay was due chiefly to the fact that a satisfactory
method had to be developed before the samples of low vita
min D potency could be assayed. However, there appeared
to be no danger of a loss of antirachitic potency because some
of the older samples were assayed 30 months after the first
test was completed, the results indicating that vitamin D in
milk fat is stable for at least 30 months when the samples are
stored at 0 °F . in the dark.
Practically all of the results given in tables 2, 3, 4 and 5
were obtained by using the alcohol extraction method, al
though most of the summer samples and a few of the more
potent winter samples were also assayed by feeding the
original fat.
d is c u s s io n
In this investigation two assumptions were made which
appear to be justifiable. It was assumed that all of the anti
rachitic potency of cows’ milk is present in the milk fat and
that there was no significant loss in potency incidental to the
separating and churning of the cream.
The standard curative feeding technic was selected for the
bioassays because this method has a number of definite ad
vantages over the prophylactic procedure. Besides the fact
hat the former is much more widely used, it permits the
VA R IA TIO N
OF V IT A M IN
D IN
M IL K
547
feeding of relatively large amounts of fat without interfering
with the test itself. In the prophylactic method the addition
of vitamin D free fat to the basal rachitogenic diet will of
itself cause a definite increase in the ash content of the bones,
the increase depending on the amount of fat added. In this
connection the slight modification of the rachitogenic diet
seems justified because the rats attained a slightly larger size
at the end of the preliminary period and had somewhat better
appetites. This permitted the feeding of larger amounts of
fat which was necessary in the case of the samples of lower
potency. Nevertheless there were limitations in the capacity
of the rachitic rats to consume fat and this necessitated con
centration of the vitamin D. The alcohol extraction method
described above seemed to solve this difficulty.
Inasmuch as Kon and Booth ( ’34) had felt that the vitamin
D in winter milk might be different from that in summer milk
because of the difference in stability to saponification, some
of the more potent winter samples were assayed both by feed
ing the original fat and an equivalent amount of the extract.
Although many of the rats failed to consume the larger doses
of fat during the first 8 days of the experimental period, a
sufficiently large number was used so that an assay at the
10-gm. level was made possible. The results indicated that
there was no apparent loss in vitamin D in making the alcohol
extractions of the winter samples tested, and served as the
basis for the assumption that the assays of fats of still lower
vitamin D potency by this method of concentration might be
reliable.
The results obtained demonstrate that milk produced by
cows managed under practical farming conditions varied as
much as 900 per cent in antirachitic potency, reaching a maxi
mum from June to September and beginning with October,
declining rapidly to a minimum which usually occurred in
February. From the assays made on the milk fats it was
calculated that the maximal potency of the milks examined
in this study was 43.8 U.S.P. units per quart. Values of 20
to 30 units per quart were not uncommon during the summer
548
H . E. BECH TEL AND
C. A . H O P P E E T
months whereas values of 8 units and less were frequently
observed during the winter months. These results in a gen
eral way corroborate those of other investigators.
Regarding the factors which contribute to the variability
in the vitamin D content of milk, the amount of exposure of
the cows to sunlight probably plays the major role. This is
strikingly indicated by the excellent correlation between the
vitamin D potency of the milk and the amount of available
sunshine as shown in figures 1, 2 and 3. Undoubtedly even
better correlation might have been obtained if a record had
been kept of the hours of actual exposure to sunlight as well
as of the ultraviolet intensity of the sunlight. The lack of
agreement during February is to be explained on this basis.
It follows from the above that the vitamin D contained in
ordinary dairy feeds, particularly roughages and silage, how
ever important this source may be to the general well being
and productiveness of the dairy cow, contributes relatively
little to the vitamin D content of the milk. Furthermore the
rapid drop in the antirachitic potency of milk which follows
the decrease in exposure of the cows to sunlight suggests that
under ordinary conditions of management and feeding the
dairy cow has practically no opportunity to build up a reserve
of vitamin D during lactation.
In comparing the Holstein and Guernsey samples as shown
in tables 2 and 3 it is interesting to note that there was little
difference in the antirachitic potency of the milk fat. How
ever, because of the higher per cent of fat in the milk of the
latter breed, the calculated vitamin D content of the milk was
greater.
V A R IA T IO N OF V IT A M IN D I N
M IL K
549
SUMMARY
1. A method is presented for the concentration of the anti
rachitic factors in milk fat thus making possible the biological
assay of fats of low potency.
2. The monthly assay of milk fats from several sources over
a period of 2 years shows that milk may vary as much as
900 per cent in antirachitic potency. Highest values were
obtained during July, August or September and lowest usu
ally in February. Vitamin D values ranging from 4.8 to
43.8 IJ.S.P. units per quart of milk were observed in the case
of Guernsey milk whereas the extreme values for Holstein
milk were 3.1 to 27.7 U.S.P. units per quart.
3. The close correlation between the antirachitic potency of
milk and the amount of available sunshine indicates that the
exposure of cows to sunlight is the major factor contributing
to the vitamin D content of milk.
4. Apparently the cow has little or no opportunity to store
vitamin D during lactation under ordinary dairy management
conditions.
LITERATURE CITED
M. H. R o sc o e 1 9 2 6 Influence o f diet and sunlight u p o n the
amount of vitamin A and vitamin D in the milk afforded by a cow.
Biochem. J., vol. 20, p. 632.
D u t c h e r , R. A. a n d H. H o n e y w e l l 1927 Feeding experiments with rats
using butters furnished by Dr. J. S. Hughes of Kansas. Penna. Agr.
Exp. Sta. 40th Am. Rep. Bui. 218, p. 4.
K o n , S . K. a n d R. G. B o o t h 1934 The vitamin D activity of butter. III. An
attempt to elucidate the nature of the labile factor in butter anti
rachitic for the rat. The antirachitic potency of lard, olive oil, egg oil
and the fatty acids of butters and lard. Biochem. J., vol. 28, p. 121.
K r a t jss , W . E. 1933 Personal communication.
L u c e , E. M. 1924 The influence of diet aud sunlight upon the growth-promoting
and antirachitic properties of the milk afforded by a cow. Biochem.
J., vol. 18, p. 716 and p. 1279.
M it c h e l l , J. McK., J. E i m a n , D. V . W h i p p l e a n d J. S t o k e s 1932 The pro
tective value for infants of various types of vitamin D fortified milk.
Am. J. Pub. Health, vol. 22, p. 1220.
R u s s e l l , W. C. 1933 Increasing the vitamin D content of milk. N. J. Agr.
Exp. Sta. Circ. 285.
Ch i c k ,
H.
and
S t e e n b o c k , H -, E . B . H a r t , B . M . R i i s i n g , C. A . H o p p e r t a n d S . B a s h e r o w
1930 Fat-soluble vitamins X X V III. The antirachitic value of cows’
milk as modified by exposure of the cow to sunlight and to radiations
from a quartz mercury vapor lamp. J. Biol. Chem., vol. 87, p. 103.
THE ANTIRACHITIC ACTIVITY OF VARIOUS PARTS OF THE
CORN PLANT AT THE TIME OF ENSILING
H. E R N E ST B EC H T E L A N D C. A. H O P PE R T
SEC T IO N S OF DAIR Y H U SB A N D R Y A N D CHEM ISTRY
P rev io u s w o rk a t this S ta tio n d em o n strated th a t corn silage has
a n tira c h itic p ro p e rtie s w hen fed to dairy calves, and to ra ts (1). The
p re se n t re p o rt is a co n tin u atio n of this w ork, and is concerned w ith the
v itam in D c o n te n t of several p a rts of the corn p lan t a t th e tim e of
ensiling'.
As indicated elsew here (1), corn is usually h arv ested for silage when
som e p a rts of th e p lan t have becom e d ry in th e field, w hile o th er p o r
tio n s of th e p lan t rem ain green. T he available d ata su g g est th a t corn
silage m ay derive its a n tira c h itic p ro p erties from th e sun-dried p o r
tions of the corn p lan t w hich are p re sen t at th e tim e of ensiling.
Experimental
In 1933, several p a rts of th e corn plant w ere collected from a field
of corn on th e day th a t th e m ateria l w as b eing ensiled. A m ong these
w ere tassels, silk, som e of the upper green leaves, and som e of the low er
dry leaves. E ach of these m ateria ls w as placed im m ediately in a d ry
ing room a t 50°—60° C. a fte r collection, and a fte r about one w eek the
dried m ateria l w as g ro u n d and sto red in stoppered bo ttles a t room te m
p e ra tu re s u n til needed for assaying.
T he vitam in D d eterm in a tio n s w ere m ade by the biological assay
m ethod. T he fo llow ing w as used as the basal rachitogenic d iet:
Y ellow corn m eal
O at m eal ...............
W h e at g lu ten
Calcium ca rb o n ate
Sodium C hloride .
D ried y e a s t ...........
38.0
37.5
20.0
3 .0
1.0
0.5
per
per
per
per
per
per
cent
cent
cent
cent
cent
cent
T h e cu rativ e technique w as used hi accordance w ith stan d ard linete s t p rocedure and vitam in potencies w ere determ ined in term s of
U. S. P. ra t u n its of vitam in D. T he m aterials to be assayed w ere
in co rp o rated a t various levels in 40 gm . of the basal rachitogenic diet
and fed to stan d ard rachitic rats. The re su lts of all assays w ere com
pared w ith th o se from ra ts w hich received a know n am ount of vitam in
D in th e form of in te rn a tio n a l reference cod liver oil.
(1) B e c h te l, H . E r n e s t ; H u f f m a n , C. F . ; D u n c a n , C. W . ; a n d H o p p e r t , C. A.
V i t a m i n D S tu d ie s in C attle. IV. C orn Silage as a S o u rc e of V i t a m i n D for D a ir y
C a ttle. J o u r n a l of D a i r y Science, in P re ss .
R e p rin t fro m M ic h ig a n A g ric u ltu ra l E x p e rim e n t Statio n Q u a rte rly
F e b r u a r y , 1936.
B u l l e t i n , V o l . IS. N o . J,
R esu lts
In Table 1 are given the an tirach itic values of the various p a rts of the
corn p lan t assayed.
Table I.
Portion of the Corn P lant A ssayed
V itam in D U nits
per Pound of
D ry M atter*
1226
2449
2449
♦Air dry basis.
+*Ten gram s had no antirachitic action w hen fed to a standard rachitic rat.
Discussion and Summary
B iological assays of the corn plant collected a t th e tim e of ensiling
d em o n strated th a t those p arts of the plant w hich w ere su n -d ried in
the field w ere p o ten t sources of vitam in D. Of p ractical im p o rtan ce
w ere th e dry leaves on the low er portions of th e plant. It w as found
th a t 0.5 gm . of the dry leaves produced a n arro w continuous line of
calcification across the m etaphyses of the radius and ulna in a stan d ard
rach itic ra t. H ow ever, as m uch as 10 gm . o f'd rie d m ateria l prep ared
from th e g reen leaves possessed no dem onstrable q u an tity of v itam in D.
T hese findings are in ag reem en t w ith o th e r w o rk in th e lite ra tu re
which show s th a t fresh, g reen plant m aterials are o rd in arily devoid of
an tira ch itic activity, w hereas plant m aterials allow ed to dry in direct
sunshine develop an tira ch itic qualities.
T he d ata presen ted in T able I. indicate th a t corn silage derives a n ti
rachitic qualities from those p a rts of the corn plant w hich w e re sundried in th e field p rio r to ensiling.
Conclusion
T he an tirach itic activ ity of corn silage is derived chiefly from those
p o rtion s of the corn plant which w ere sun-dried in th e field p rio r to
ensiling.
R ep rin ted from J o u r n a l
of
D
a ir y
S c i e n c e , Ju n e, 1936, V ol. X I X , N o . 6
V IT A M IN D S T U D IE S IN CATTLE
IV . C o r n S i l a g e a s a S o u r c e o f V it a m in D f o r D a ir y C a t t l e *
H. ERNEST BECHTEL,** C. E. HUFFM AN, C. W. DUCAN
a n d C. A. HOPPERT
Sections of D airy Husbandry and of Chemistry, Michigan Agricultural Experiment
Station, E ast Lansing
I n previous reports from this Station, experim ental d ata have been sub
m itted upon the an tirachitic value of sun-cured hay for d airy cattle (1),
the vitam in D sparing action of certain magnesium compounds when only
small am ounts of this vitam in were present in the dairy ration (2), and
the essential n atu re of ra d ia n t energy in the dietary regime of the dairy
calf (3). The present report is concerned with the antirachitic value of
corn silage fo r d airy cattle.
Hess and W einstock (4) reported th a t green plants grown in the dark
contained no vitam in D but became antirachitic afte r ultraviolet irra d ia
tion. Bethke, K en n ard and K ik (5) failed to prevent leg weakness in
chicks when green, fresh red-clover was fed as 50 per cent of the ration.
Steenbock and associates (6) found th a t clover hay cured in the dark was
inactive antirachitically. M ellanby and K illick (7) reported th a t summergrown grass contained more of the calcifying factor th an cabbage. G-reen
spinach grown in m idsumm er has been reported by Chick and Roscoe (8)
to have a slight b u t appreciable antirachitic value. W hile an investigation
regarding the actual feeding of fresh, green plants as a source of vitam in D
to dairy cattle is lacking, the indications are th a t fresh, green pasture
grasses, in general, are poor sources of antirachitic substances.
As previously indicated (1), the antirachitic potency of hays is related
to th eir exposure to solar ultraviolet rays. I t is the usual practice to harvest
corn fo r silage when m any of the ears have become dented, the bottom three
or fo u r leaves and a portion of the husk have become d ry while the rem ain
der of the p lan t is still green. Corn at this stage of m atu rity is m ostly green
plant substance and for this reason has been considered a poor source of
vitam in D, although this point has not been specifically investigated.
Because of the presence of sun-dried leaves, however, and other m aterial
* Published with the permission of the Director of the Experiment Station as Journal
Article No. 250 (n.s.).
** This manuscript is part of a dissertation presented to the Faculty of the Graduate
School of Michigan State College in partial fulfillment o f the requirements for the degree
of Doctor of Philosophy.
Received for publication January 27, 1936.
359
360
BECHTEL* H U F FM A N ; DUNCA N AND HOPPERT
on the p lan t a t the tim e of ensiling, it was thought possible th a t corn silage
m ight have an appreciable antirachitic value.
The object of this investigation was to determ ine the an tira ch itic value
of corn silage for dairy cows.
E X P E R IM E N T A L
The corn silage was made from dent corn which was cu t w hen the
kernels were dented and the lower leaves were dry. One sam ple of silage
was taken fo r each of the following years, 1931-1934, inclusive, fo r biolog
ical assay w ith rats. Each sample of silage was im m ediately p u t in a d ry in g
room a t 50°-60° C. afte r collection and afte r about one week, th e dried
m aterial was ground and stored in stoppered bottles at room tem p eratu res
u n til needed fo r assaying. The vitam in D content of the silage was d eter
m ined by the curative feeding technique w ith ra ts according to a sta n d a rd
line-test procedure. The vitam in D contents of the silage, in term s of
U. S. P. ra t units, are shown in Table 1.
TABLE 1
Vitamin D content o f silage, U. S. P. rat im its
U N IT S PER
P O U N D DRY
M ATTER*
C U R A T IV E M A T E R IA L
Corn
Corn
Corn
Corn
silage
silage
silage
silage
1931
1932
1933
1931
................................................
................................................
................................................
................................................
165
122122
165
* Air dried basis.
The nineteen grade-H olstein dairy calves of either sex w hich were used
in this experim ent were divided into 5 groups. The calves in the first 4
groups were placed on experim ent at b irth b u t the calves in group 5 were
several m onths of age when placed on this experim ent. The m anagem ent
of the calves and the composition of the rachitogenic calf ratio n were sim ilar
to th a t previously reported (1 -3 ). Blood samples were obtained from each
of the experim ental calves every two weeks and the plasm a from each sam ple
was analyzed fo r calcium, inorganic phosphorus (9) and m agnesium (10)
by methods already recorded. A t the tim e of post-m ortem exam ination,
certain bones were saved from each anim al and studied histologically (11).
The chemical analysis of the various feeds used in this investigation are
given in Table 2.
The calves in Group I, C-140, C-161 and C-164, were fed th e u n su p p le
m ented basal rachitogenic ration. The calves in G roup II, C-135, C-139
and C-141, each received the ash from one pound of d ry silage p e r day in
addition to the basal ration. The calves in Group I I I , C-132, C-136, C-137,
361
VITAMIN D STUDIES IN CATTLE
TABLE 2
Chemical analysis of feeds and water
M A T E R IA L
per cent
Whole milk ......................
Skim milk ...........................
Corn and oats ..................
Grain mixture ..................
Corn silage 1931-32* .....
Corn silage 1932-33 ........
Corn silage 1933-34 ........
Corn silage 1934—35 ........
Corn silage ash ................
Water .................................
Ca
M O IS T U R E
11.80
11.30
76.61
78.02
70.51
69.00
per cent
0.120
0.122
0.128
0.494
0.175
0.101
0.103
0.126
7.190
0.0082
P
Mg
per cent
0.093
0.096
0.321
0.364
0.062
0.054
0.054
0.064
2.950
per cent
0.012
0.012
0.156
0.199
0.122
0.067
0.088
0.115
6.770
0.0027
!
* Each year’s analysis began with September.
C-138 and C-145, received from one to six pounds of silage per day per
animal. Following the above curative feeding trials, the calves in Group IV,
C-156, C-159 and C-160, were used in a preventive trial. The calves in
this group received the basal ration but corn silage was added as the sole
source of vitam in D when the calves were 30 days of age. The calves in the
above groups which survived were slaughtered at approxim ately 190 days of
age, w ith the exception of C—159 which was changed to another experim ent.
Calves C-168, C-169, C-188, C-195 and C-167 were subsequently added
to this experim ent as Group V. . The first four calves were rachitic a t the
time when silage was added to th eir basal rachitogenic ration. Calf C-168
was 269 days of age and had been suffering from rickets for about 50 days,
calf C-169 was 348 days of age and had been suffering from rickets for
about 100 days, calf C-188 was 428 days of age and had had rickets for more
th an 200 days and calf C-195 was 395 days of age and had had rickets for
at least 60 days when silage was added to their rations. Calves C-168 and
C-169 were subsequently slaughtered, C-188 died while on experim ent and
C-195 was continued on experim ent to determ ine the effect of the ration
upon grow th and reproduction. Calf C—167 was norm al when silage was
added to the ration.
R E SU L T S
The results obtained from the experim ent are presented in Tables 3-7,
inclusive. Table 3 summarizes the data pertaining to the age when the
calves were placed on experim ent, the age when the supplem ent was added,
the first evidence of rickets, the age of the calves at the term ination of the
experim ent and the term inal plasm a calcium, magnesium and inorganic
phosphorus values. Table 4 gives the data for a representative animal in
Groups I-IV , inclusive.
362
BECHTEL, H U F F M A N , D U N C A N AND HOPPERT
TABLE 3
Summary of data pertaining to calves which received the rachitogenic ration
Calf
no.
C-140
C-161
C-164
EXPER.
STA RTED,
AGE
R A C H IT IC .
AGE
SUPPLEM ENT
ADD ED . AGE
days
days
73
127
108
Group II.
0135
0139
0141
0132
0136
0137
0138
0145
1
1
1
1
1
1
1
1
Group IV.
0156
0159
0160
1
1
1
192
269
348
428
366
88 D 1
162 D
158 D
9.4
9.3
8.8
| Mg
p
mg. per 100 cc.
13.80
9.06
6.69
2.33
2.19
1.87
6.51
8.62
5.84
2.80
1.68
2.43
Basal ration plus silage ash
93
90
79
72
71
73
Group III.
Basal ration plus silage
93
95
92
86
92
93
81
67
86
92
108 D
114 D
193 S2
193
191
192
193
190
S
S
S
S
S
6.8
7.3
8.4
8.2
8.6
8.6
10.0
10.0
3.81
3.68
3.50
7.35
2.27
2.00
2.45
2.82
2.45
Basal ration plus silage before the onset of rickets
32
33
30
Group V.
0167
0168
0169
0188
0195
T E R M IN A L P L A SM A
Ca
days
days
Group I. Basal ration
none
>i
}>
1
1
1
EXPER.
ENDED.
AGE
192
269
348
428
366
161
116
95
186 S
190s
193 S
7.5
3.63
1.56
8.0
3.01
1.80
_
Basal ration plus silage (older calves)
none
210
248
225
300
11404
371 S
414 S
520 S
813“
11.9
12.3
8.5
7.27
6.79
4.13
1D denotes died. 2 S denotes slaughtered. 3 Changed to another experiment.
on experiment. * Changed to another experiment, cow still alive.
3.12
2.45
2.33
* Still
Group I .— The three calves in this group received the unsupplem ented
basal ration. A norexia was m anifested by C-140 and C-161 a fte r th e onset
of rickets and C-161 also had a convulsion a t 162 days of age. C -164 h a d a
severe convulsion at 128 days of age at which tim e 5 cc. of parathorm one
was injected subcutaneously. A m ild convulsion was again noted a t 157
days of age and the calf was found dead on the follow ing day. T he condi
tion of the pen indicated th a t the calf had died in a convulsion.
Group I I .— The three calves in this group h ad th e ir basal rachitogenic
ration supplem ented w ith silage ash. C-135 contracted b ilatera l pneum onia
363
VITAMIN D STUDIES IN CATTLE
H
03
3 bl
r$
02i
p?
H C3 CO tO O CO
0 3 Q M H 0 0 rH
0 3 0 3 0 3 0 3 rH 0 3
0 3 rH rH rH 0 3 0 3
03 03 03 03
0 3 0 3 0 3 C3 H
H
H
.-2
na
a> o5
CO 1C
1C 0 3
CO* CO*
1C 0 3 b - 0 0
O i H j CO 0 3
b-*t > 1C t >
S rP
C O fllO O O H H i
CO 0 0 CO 0 0 rH 0 0
1C CO CO 1C 1C i d
.§ P
CD CD 1C b ’ 1C CD IQ
rP
tuo«
9 9
d
O
Growth data, mineral and silage intalces and blood plasma values of representative calves
5q |
*
rH CO ©
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0 5 0 0 b 4 CD b - 0 0
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W r j N O 05 00 r j
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c q H j rH c o c o
0 3 0 0 0 5 03
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c d b 4 0 5 o cd t 4
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r—I
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CO 0 0 0 0 t> c*
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d £g
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f^s* O CO CD 0 5 0 5
rH rH H rH H
00 05 N O
0 0 CO b - 0 0
rH 0 3 0 3 0 3
0 3 CO 0 3
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H H rH rH 0 3 0 3 0 3
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364
BECHTEL, H U F F M A N , D U N C A N AND HOPPERT
and died at 108 days of age. C—139 had a convulsion a t 88 days of age and
died at 114 days of age. C—141 had convulsions at 129 and 167 days of age.
P arathorm one was injected afte r each convulsion. Convulsions were again
noted at 183 and 184 days of age. ’ The calf was slaughtered a t 193 days
of age.
Group I I I — A fter 90 days of age, the five calves in this group received
the basal ration supplem ented w ith small am ounts of silage. A ll of the
calves showed the clinical evidences of rickets from 62 to 93 days of age.
C-137 had convulsions at 83 and at 90 days of age. The corn silage supple
m ent was started at 92 days of age and convulsions were observed at 128
days of age. The calves in this group were slaughtered at approxim ately
190 days of age. C-145 was representative of this group, therefore, the
calcium, phosphorus and magnesium intakes and the concentration of these
constituents in the blood plasm a are presented in Table 4.
Group I V .— The three calves in this group had th e ir ratio n supplem ented
w ith corn silage at about 30 days of age and before the onset of rickets.
The calves did not develop rickets un til they were from 95 to 161 days of
age. They did not consume enough silage a t an earlier age to prevent
rickets b u t the silage consum ption tended to delay the sym ptoms. As they
became older an insufficient am ount of silage was consumed to prevent
rickets because of anorexia. C—156 and C—160 were slaughtered a t about
190 days of age and C-159 was changed to another experim ent a t 210 days
of age. The representative experim ental d a ta secured from C-159 are
presented in Table 4.
Group V .—W ith the exception of C-167, the older calves w hich were
used in this group to determ ine the antirachitic effect produced by the
ingestion of large am ounts of silage were rachitic when placed on experim ent.
C alf C-167. This calf had previously been used to determ ine th e efficacy
of solar ultraviolet radiation in preventing the m anifestations of rickets (3).
Table 5 presents the growth data, feed consum ption and blood plasm a values
from 210 days of age un til the term ination of the experim ent. She received
corn silage as the sole source of vitam in D at 192 days of age an d ate it w ith
avidity from the very beginning, grew norm ally and m aintained a sleek
appearance at all times. C-167 was first bred a t 467 days of age and again
at 487 and 515 days of age. She aborted a t 601 days of age an d was again
bred at 662 and 765 days of age. B eginning at 920 days of age, all grain
was w ithheld from the ration and corn silage was the sole source of food
other than wood shavings, salt and w ater. This heifer gave b irth to a norm al
85-pound heifer calf at 1032 days of age and had a retain ed placenta. C-167
a\erag ed approxim ately 45 pounds of milk p er day, containing 3.5 p er cent
fat, during the first 100 d aj s of lactation on a ratio n w hich consisted of the
rachitogenic grain m ixture, corn silage, wood shavings, salt and w ater. Pre-
365
VITAMIN D STUDIES IN CATTLE
M g
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366
BECHTEL, H U F F M A N , D U N C A N AND HOPPERT
lim inary results indicate th a t the m ilk produced by C—167 contained con
siderably less th an 2.7 U. S. P. vitam in D u n its p e r q u art. The blood plasm a
data fail to reveal any significant variations from norm al. H ow ever, there
were two grad u al drops in the inorganic phosphorus values. The first drop
was associated w ith the n a tu ra l physiological disturbance due to abortion
and the second was due to p artu ritio n .
C alf C -168. This calf received from 15 to 20 pounds of corn silage per
day as the sole source of vitam in D from 269 days of age u n til the end of the
experim ent. The calf was suffering from severe rickets w hen first placed on
experim ent and the blood calcium and inorganic phosphorus values were
7.6 and 9.84 mg. per 100 cc. of plasma, respectively. The silage was readily
consumed from the beginning and at 303 days of age the calf was able to
rise to its feet w ith much less difficulty th an a week earlier. T he plasm a
calcium and inorganic phosphorus values were 9.3 and 5.48 mg. a t 303 days
of age and by the following week they had re tu rn e d to norm al, 10.9 an d 6.76
mg., respectively. The gains in body w eight were also g re ater fro m this
tim e on u n til the anim al was cured of rickets and slaughtered a t 371 days
of age. The term inal calcium and inorganic phosphorus values w ere 11.9
and 7.27 mg. The kidneys showed extensive areas of scar tissue.
C alf C-169. This calf received 15 pounds of corn silage p e r day as the
sole source of vitam in D beginning a t 348 days of age. I t was severely
rachitic and had difficulty in rising to its feet. The blood calcium an d inor
ganic phosphorus values were 8.0 and 5.84 mg. per 100 cc. of plasm a. The
corn silage was readily consumed and 10 days la te r the calf was able to rise
to its feet w ith much less difficulty. The blood values were approaching
th eir norm al concentrations by 370 days of age and the calf began to gain in
body weight. C-169 was cured from rickets when slaughtered a t 414 days
of age and the term inal calcium and inorganic phosphorus values were 12.3
and 6.79 mg. The kidneys showed more extensive areas of scar tissue th an
observed in C-168. The d ata are tabulated in Table 6.
Calf C—188. This calf was 428 days of age and h ad been suffering from
rickets fo r more th an 200 days when corn silage was added to the ra tio n as
the sole source of vitam in D. The anim al was severely rach itic a t this tim e
and was only m aintaining a constant body weight. The blood calcium and
inorganic phosphorus values were 7.2 and 7.06 mg. p e r 100 cc. of plasm a.
The silage was readily eaten b u t the grain was refused p a r t of th e time.
By 472 days of age the anim al was extrem ely stiff and lam e and was ra re ly
seen standing so th a t it became necessary to place the feed in a basket on the
floor of the stall. The plasm a calcium had now increased to 7.8 mg. b u t the
inorganic phosphorus had declined to 4.51 mg. P lacing the feed before the
anim al resulted in an increase in silage consum ption b u t no im provem ent
was noticed in its well-being. A t 504 days of age the anim al was observed
367
VITAM IN D STUDIES IN CATTLE
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BECHTEL, H U F F M A N , DUN C A N AND HOPPERT
struggling to its feet, breathing h ard and w ith nostrils distended.
The
following week C—188 was emaciated, unable to rise to its feet a n d refused
all feed and water. The plasma calcium and inorganic phosphorus values
at this time had declined to 7.1 and 2.44 mg. The animal lapsed into a coma
tose condition at 518 days of age and failed to respond appreciably either
to intravenous injections of solutions of calcium gluconate, glucose an d m ag
nesium sulfate or to a subcutaneous injection of viosterol. D eath occurred
at 520 days of age. There were patches of scar tissue in the kidneys, the
muscle tissue showed considerable edema and there were evidences of muscle
in ju ry around the leg joints. The most characteristic autopsy finding was
the generalized condition of pitting and erosion of the artic u la r surfaces of
the long bones. Illustrations and a detailed description of the histological
findings are given elsewhere (11).
Calf C-195. This calf was 366 days of age and had suffered from rickets
for more than 60 days when corn silage was added to th e rachitogenic ration
as the sole source of vitamin D. The an im al’s legs were stiff and the knees
were bowed markedly.
The plasma calcium and inorganic phosphorus
values were 9.5 and 3.01 mg. per 100 cc. at this time. The appetite for
silage was only fa ir but it gradually improved and by the following month
the calf was able to rise to its feet with much less effort. The addition of
silage had little effect upon the blood constituents d u rin g the first two
months b ut during the next two months, the concentration of calcium
returned to normal and the inorganic phosphorus manifested a slow but
steady rise. E stru s was first noted at 413 days of age, at which tim e the
bowing of the knees had become less marked. Im provement was ra p id after
that time. C-195 had recovered from rickets by 543 days of age, was bred
and thereafter made normal gains in body weight. The heifer gave birth
to a normal 82-pound bull calf at 813 days of age. The d a ta secured from
this animal are summarized in Table 7.
DISCUSSION
W hen calves less than 190 days of age were used as test animals it was
impossible to either cure or prevent rickets by supplem enting the basal
rachitogenic ration with corn silage. A larger percentage of the calves
survived until 190 days of age, however, when silage supplem ented the basal
ration (Table 3). This suggested that silage exerted some antirachitic activ
ity although the amount ingested was insufficient to m ain tain or to restore
health. H igher intakes of silage were precluded by anorexia. Calf C-145
had the best appetite for silage b ut the average consumption was less th an
six pounds per day. This amount of silage was equivalent to a daily intake
of approximately 5 grams of silage dry-m atter p er kilo of body weight and
was ineffective in curing rickets. The ingestion of three pounds of corn
silage per day failed to prevent rickets in young calves.
369
VITAM IN D STUDIES IN CATTLE
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BECHTEL, H U F F M A N , DUN C A N AND HOPPERT
W hen older calves were used, the clinical manifestations of rickets were
alleviated in three out of four eases by the daily ingestion of 15 to 20 pounds
of silage (Tables 6—7). Backets was also prevented in C—167 from six
months to three years of age by the ingestion of corn silage (Table 5). The
abortion of C-167 was probably not associated w ith a vitam in D deficiency
in view of the results reported by Moore and associates (12). The decrease
in the concentration of inorganic phosphorus in the plasm a of C—167 and
C-195 at the time of parturitio n and the abortion of C-167 was associated
with the physiologic disturbances caused by these acts ra th e r th a n by a
deficiency of the antirachitic factor. I t has been shown (9) t h a t such a
change in the composition of the blood plasma is not uncommon in dairy
cattle at the time of parturition.
I n order to compare the results obtained with the various animals it
seems convenient to express the intake of corn silage in term s of gram s of
silage dry-matter per kilo of body weight. On this basis, 7 to 10 gm. of
silage dry-matter per day was not only effective in curing and preventing
rickets but also supplied a sufficient amount of the antirachitic factor for
normal growth and reproduction.
Calf C-188 ingested from 15 to 20 pounds of silage p er d ay which was
equivalent to 8 to 12 gm. of silage dry-m atter per kilo of body weight, yet
failed to recover from rickets. This was the only one of the older calves
which failed to respond favorably to silage feeding. This calf is regarded
as an atypical case. The inability to eure C-188 is ascribed to the severity
of the rachitic condition which had developed before curative measures were
begun. F ailure to utilize its feed properly, together with its emaciated
appearance and autopsy findings in the joints, indicate th a t the animal was
suffering from far-advanced rickets which had given rise to other n u tr i
tional disturbances refractory to ordinary vitam in D therapy.
H u ty ra
and Marek (13) state th at if rickets is fa r advanced and has given rise to
emphatic nutritional disturbances, death always follows, either through
exhaustion or through some complication. The ultim ate healing of the
process is frequently prevented by ulcers formed in the course of the disease
in the articular cartilages.
Bioassays made with rats indicated th a t 7.5 to 10.0 gm. of d ry corn silage
contained an equivalent of 2.7 U. S. P. units of vitam in D (Table 1).
Assuming th a t 10.0 gm. of dry silage contained 2.7 U. S, P. r a t units, d uring
the period 1931-1934 inclusive, the daily intake of vitam in D un its was
calculated for each of the experimental calves (Tables F-7).
The representative tables included in this pap e r show t h a t the addition
of corn silage to the rachitogenic ration appreciably increased th e intake
of magnesium. In view of the results of a recent investigation (2), it is
possible th at the presence of magnesium in silage m ay augm ent the efficacy
of the antirachitic material in corn silage.
VITAMIN D STUDIES IN CATTLE
371
SUM M ARY AND CONCLUSIONS
1. I t was the purpose of this investigation to determine the antirachitic
value of corn silage for dairy calves by the use of both curative and p re
ventive feeding trials.
2. Samples of corn silage for the years 1931-1934 contained from 122 to
165 U. S. P. vitamin D units per pound of dry matter.
3. Calves less than 190 days of age were unsuitable test animals for both
curative and preventive trials because of anorexia and failure to ingest
adequate amounts of silage. In one instance, the feeding of corn silage to a
yearling calf failed to cure severe rickets complicated with muscle atrophy
and erosion of the articular surfaces.
4. The daily ingestion of an equivalent of 7.0 to 10.0 gm. of dry corn
silage per kilo of body weight was effective in curing and preventing rickets
in yearling calves and also supplied sufficient antirachitic material for
normal growth and reproduction in dairy cows.
5. When corn is cut at the usual stage of m aturity for corn silage it
possesses definite antirachitic qualities when fed to dairy cows.
The writers are indebted to Mr. C. C. Lightfoot for technical assistance
in the determination of the blood values and to Mr. 0. B. W inter and Miss
Lillian I B utler for the chemical analyses of the feeds.
LITERATURE CITED
1. H o f f m a n , C. F., a n d D u n c a n , C. W. Vitamin D studies in cattle. I. The anti
rachitic value of hay in the ration of dairy cattle. J o u r n a l of D a ir y S c ie n c e
18: 511. 1935.
2.
, A N D ------------------- . Vitamin D studies in cattle. II. The vitamin
D s p a r in g a c tio n o f m a g n e s iu m in th e r a t io n o f d a ir y c a t t le .
3.
4.
5.
6.
7.
8.
J o u r n a l of D a ir y
18: 605. 1935.
D u n c a n , C. W., a n d H u f f m a n , C. F. Vitamin D studies in cattle. III. Influence
of solar ultraviolet radiation upon the blood chemistry and mineralmetabolism
o f dairy calves. J o u r n a l of D a ir y S c ie n c e , 19: 291, 1936.
H e s s , A. F ., a n d W e i n s t o c k , M. Antirachitic properties imparted to inert fluids
and to green vegetables by ultraviolet radiation. J. Biol. Chem. 62: 301. 1924.
B e t h k e , R. M., K e n n a r d , D. C., a n d K i k , M. C. Nutritional studies of the growing
chick. I. Relation of sunlight and green clover to leg weakness in chicks. J.
Biol. Chem. 63: 377. 1925.
S t e e n b o c k , H., H a r t , E. B., E l v e h j e m , C. A., a n d K l e t z i e n , S. W. F. D ie t a r y
factors influencing calcium assimilation. VI. The antirachitic properties of
hay as related to climatic conditions with some observations on the effect of
irradiation with ultraviolet light. J. Biol. Chem. 66: 425. 1925.
M ellAnby, M., and K illicic, E. M. CXIII. A preliminary study of factors influenc
ing calcification processes in the rabbit. Biochem. J. 20: 902. 1926.
C h ic k , H., a n d R o s c o e , M. H . The antirachitic value of fresh spinach. Biochem. J.
20: 1. 1926.
S c ie n c e
372
BECHTEL, H U F F M A N , D U N C A N AND HOPPERT
9. H u f f m a n , C. F ., D u n c a n , C. W., R o b i n s o n , C. 8 ., a n d L a m b , L . W. Pliosphorus re
quirement of dairy cattle when alfalfa furnishes the principal source o f protein.
Mich. Agr. Exp. Sta. Tech. Bull. 184. 1933.
10. D u n c a n , C. W., H u f f m a n , C. F., a n d R o b i n s o n , C. S. Magnesium studies in calves.
I. Tetany produced by a ration of milk or milk with various supplements. J.
Biol. Chem, 108; 35. 1935.
11. B e c h t e l , H . E., H a l l m a n , E. T., H u f f m a n , C. F., a n d D u n c a n , C. W, Pathology
of rickets in dairy calves. Mich. Agr. Exp. Sta. Tech. Bull. 150. 1936.
12. M oore , L. A., H u f f m a n , C. F., a n d D u n c a n , C. W. Blindness in cattle associated
with a constriction of the optic nerve and probably of nutritional origin. J.
Nutrition 9: 533. 1935.
13. H u t r y a , F., a n d M a r e k , J. Special Pathology and Therapeutics of the Diseases
of Domestic Animals. Alexander Eger, Chicago 3: 244. 1926.
T echnical B ulletin No. 150
May, 1936
PATHOLOGY OF RICKETS IN
DAIRY CALVES 7
H.
E R N E S T B E C H T E L , E. T. H A L LM A N , C. F. H U F FM A N
'' A N D C. W . D U N C A N
AGRICULTURAL EXPERIMENT STATION
MICHIGAN STATE COLLEGE
O f Agriculture and A pplied Science
SE C T IO N S O F D A IR Y H U S B A N D R Y , A N IM A L
PATH O LO G Y, A N D C H E M IST R Y
E ast Lansing, Michigan
T e c h n i c a l B u l l e t i n N o . 150
M a y , 1936
PATHOLOGY OF RICKETS IN
DAIRY CALVES
H.
ERNEST BECHTEL, E. T. HALLMAN, C. F. HUFFMAN
AND C. W. DUNCAN
AGRICULTURAL EXPERIMENT STATION
MICHIGAN STATE COLLEGE
Of Agriculture and Applied Science
SECTIONS OF DAIRY HUSBANDRY, ANIMAL
PATHOLOGY, AND CHEMISTRY
E a s t L a n s in g , M ic h ig a n
TABLE OF CONTENTS
Page
n trod u ctio n .........................................................................................................
-3
Materials and M e t h o d s ............................ ......................................................
^
lesu lts ..................................................................................................................
C haracteristic S y m p to m s of V itam in D Deficiency in C a lv e s . .
^
5
T h e N orm al C ostochondral J u n c t i o n ................................................
5
1.
Zone of R eserv e Cells
*• • •
5
2.
Zone of Cell P ro life ra tio n .............................■'...................
6
3.
Zone of Cell G ro w th ...............................................................
7
4.
Zone of M a tu re C e l l s .............................................................
7
5. Zone of C artilage R em o val ............................................... *
6.
Zone of Ossification ...............................................................
7.
Zone of Com pact S ub stance ..............................................
T h e Costochondral J u n c tio n in R i c k e t s ..........................................
1.
Zone of R e s tin g C artilage ..................................................
2.
Zone of Cell P ro life ra tio n ...................................................
3.
Zone of Cell G ro w th .............................................................
4.
Zone of M a tu re Cells ............................................................
5.
Zone of Cartilage R em o v al .................................................
6.
Zone of O s s i f i c a t i o n ................................................................
8
8
10
10
H
11
11
11
12
14
7.
Zone of Com pact S u bstance ...............................................
T he C ostochondral J u n c tio n in H e a lin g R ic k e ts ......................
R o e n tg e n o g ra p h ic A ppearan ce of N o rm a l and R ick etic R ib s.
G ro w th as a M odifying F a c to r .........................................................
C om plications in R ick ets ......................................................................
T y p e of R ickets and Plistological A lte ra tio n s .............................
D i s c u s s i o n .............................................................................................................
S u m m a r y ......................................................................................... ; ...................
Conclusions ...........................................................................................................
L ite ra tu re Cited ................................................................................................
14
15
15
16
16
17
17
19
21
22
Tables ....................................................................................................................
Illustrations .........................................................................................................
23
32
P A T H O L O G Y O F R IC K E T S IN
DAIRY CALVES
H . E R N E S T B E C H T E L , E . T. H A L L M A N , C.
F. H U F F M A N
A N D C. \ V . D U N C A N
INTRODUCTION
Recognition of the im portance of vitamin D in dairy calf nutrition
has resulted in considerable w o rk on the production and prevention
of rickets in calves by the Michigan (1-6), the Pennsylvania (7) and
the W isconsin A gricultural E xperim ent Stations (8). This report p ri
marily- concerns results of a study of the pathology of ricketic bones
produced in calves at the Michigan A gricultural E xperim ent Station.
F ew histological data are available which deal with rickets in calves.
H u ty ra and M arek (9) presented a good clinical picture of calf rickets
b u t reported nothing concerning- the histological findings, although
th e y included several general anatomical changes which w ere due to
rickets. These investigators also noted th a t the bone development of
a one-year-old child corresponded to the bone development of a calf
during- fetal life. F ro m this observation, they concluded th at calf
rickets and late rickets in children were similar. M arek and W ell
m an (10) reported considerable d a ta 'u p o n the histology of rickets in
sheep and swune but relatively few data upon calves. The Pennsylvania
station (7) presented a brief description of a photom icrograph of the
n inth costochondral junction taken from a ricketic calf. No histological
s tu d y of calf rickets was included in the report from the Wisconsin
station (8). T heiler’s (11) w ork in South Africa was concerned with
aphosphorosis in heifers betw een two and three years of age, so th a t
his photom icrographs are not directly applicable to the present study
because of the age of his animals. H arris (12) has published extensively
on hum an rickets. Maxwell, H u and Turnbull (13) have described in
detail the histology of a case of fetal rickets in the hum an being. The
histology of rickets in the ra t has been studied by Pappenheim er (14)
and Dodds and Cameron (15) ’who have stated th a t the histological
pictures in hum an and in ra t rickets are fundamentally identical. The
w o rk of the latte r group of investigators has been a helpful guide in
th e present histological study.
P au city of fundam ental data in the literatu re concerning the histol
og y of rickets in dairy calves is the justification for reporting the pres
e n t study in detail.
M ICH IGA N T E C H N IC A L B U L L E T IN NO. 150
MATERIALS AND METHODS
M o r e t h a n 1 0 0 d a i r y calves h a v e b e e n u s e d in t h e e x p e r i m e n t s o n low'
v i t a m i n D r i c k e t s w h i c h h a v e m a d e r i c k e t i c b on e s a v ai la bl e f o r this
Avork. F r o m t hi s g r o u p , five n o r m a l a n d e le ve n r i c k e t i c ca lv e s w e r e
s e le ct ed f or t h e d et ai le d h i s t ol o gi c a l s t u d y w h i c h is p r e s e n t e d in t hi s
bulletin. T h e m a j o r i t y of t h e s ele ct e d case s Avere m a l e calves o f g r a d e
H o l s t e i n b r ee di ng - a n d v a r i e d f r o m 1 5 1 t o 5 2 0 d a y s of a g e a t d e a t h .
T h e d u r a t i o n of r ic ke ts, as in di ca te d b y t h e b lood p l a s m a a n a l y s e s f o r
c alc iu m a n d i n o r g a n i c p h o s p h o r u s , v a r i e d f r o m 3 8 t o 2 3 9 d a y s so t h a t
thi s g r o u p r e p r e s e n t s m a n y s t a g e s in t h e s e v e r i t y of t h e disease.
E a rly in the investigation it becam e a p p a re n t t h a t th e co sto c h o n
dral junction at the v en tra l end of th e rib wras th e best index to th e
severity of rick ets because it showed to a g r e a t e r deg ree the sam e
changes displayed by the hum erus, femur, m etac arp u s and m e ta ta r s u s .
M id-frontal sections of about 4 mm. in thickness w e re ta k e n from
the last th ree inches of the v e n tra l end of the left eighth rib of each
calf. These sections w ere studied by the com parison of r o e n tg e n o
gram s, p h o to g rap h s of specimens stained in silver n it r a t e solution, and
histological sections.
A f t e r
c o n s id e r a b le
a d o p te d
in
th e
s p e c im e n s
ru n n in g w a s h e d
T h is
w e r e
fix ed
w a t e r ,
a
a q u e o u s
few'
h o u r s
p r o c e d u r e
c a tio n ,
o t h e r
tio n
o f
AA'hich A v a s
Avas
o s te o id
u s e d
s e c tio n s
a lso
s ta in e d
t h ic k n e s s
a c id u la te d
a
p e r
o f
a n d
p e r
u s u a l
a n d
to
lie
less
e ig h t
p e r
a lc o h o l
c e n t
a n d
m u c h
Avere
flat
a n d
s o lu tio n ,
e o sin
a c e tic
in
s ta in e d
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th e
o f
slides.
th e
T h e
o f
b y
s o lu tio n
S u c h
h e m o t o x y l i n
in
w e r e
t h r e e
u p
liver,
t h e
a t
H a r r i s ’
s e c o n d s
in
five
b y
th e
s e c
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in
a s
e o s in
t e c h
t i m e - c o n
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s e v e r a l
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m a d e
tis s u e s
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Avas
fo r
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a ls o
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T h e
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e o s in
s o lu tio n ,
of
in
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p e r
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5
a n d
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T h e
w e r e
s ta i n in g
p a ra ffin
Avas
:
2 4
a
m e t h o d
e n c o u n t e r e d
t h e
so lu tio n .
Z e n k e r ’s
in
Avas
folloA ved
50
w re e k s
m e t h o d
T h e
M o s t
s o lu tio n .
E a c h
a c id
o n
Avere
six
T h is
C e llo id in
difficulty
p e r f e c tly
to
e x c e lle n t
tis s u e s.
p e r m a n e n tl y .
fixed
C.
tis s u e s
Aveek.
e x c lu s iv e ly .
in
37°
f o u r
in
f e w
le a s t
t h e
f r o m
n itric
a
a t
a t
e v e r y
th e
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s tu d ie s
u n til
r e q u ir e d
o s s e o u s
w 'a s h e d
a lc o h o l
d e c alc ified
r e s u lte d
t e c h n i q u e
h is to lo g ic a l
f o rm a lin ,
c e n t
d i c h r o m a t e
to
fo llo w d n g
f o r
r e n e w e d
so lu tio n .
g lac ia l
Avere
p e r
a n d
m ic r o n s
c e n t
t h e
s e c tio n s
c e n t
80
w a t e r ,
a n d
h e m o t o x y l in
sp le e n
te c h n iq u e
th e
o f
o n e -h a lf
c e n t
2
p e r
in
w 'o rk
t h e
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a lm o s t
la b o rio u s ,
in
w a s
fa r
cello id in
a
10
d e c a lc ify in g - s o lu tio n s ,
s u m in g ,
tio n s
in
of
p o t a s s i u m
u s u a lly
s o lu tio n
betA veen
n iq u e
in
h a r d e n e d
s o lu tio n
c a lc ify in g
p r e l i m i n a r y
p r e p a r a ti o n
a
7 0
d r o p s
k i d n e y
p a ra ffin
s o lu tio n s
in
m a n n e r .
RESULTS
The ages of the calves at death, the d u ra tio n of the rick etic condition
and o th e r d ata p e rta in in g to the anim als used in th e study of bone
histology are sum m arized in Table 1. T h e groAvth d a ta and th e c o n
c e n tratio n s of calcium, inorganic ph osp h o ru s and m a g n esiu m of the
blood plasm a of each individual calf are p re s e n te d in Tables 2-9, in
clusive.
PATHOLOGY OF RICKETS IX DAIRY CALVES
Characteristic Symptoms of Vitamin D Deficiency in Calves
In this investigation the decrease in the concentration of calcium
a n d /o r inorganic phosphorus of the blood plasma was am ong the first
signs of low vitamin D rickets. The most conspicuous post m ortem
findings, however, were those which apparently accompanied the blood
changes and which occurred in the bones. Clinically, the skeletal changes
included bow ing of the forelegs either forw ard or to the side, swell
ing of the knee and hock joints, straig htening of the pasterns, occa
sional ring-like swellings on the pasterns, and hum ping of the back
(Fig. 1). P o ste rio r paralysis occurred in cases of fractured vertebrae.
F rac tu red femora sometimes occurred. Other symptoms frequently
observed were stiffness of gait, dragg ing of the rear feet, standing
with the rear legs crossed, irritability, tetany, rapid respiration, bloat,
anorexia for grain and roughages but not for milk, weakness and in
ability to stand for an)- length of time, and finally the retard ation or
complete cessation of g ro w th in bod)' weight.
The post m ortem examinations of the ricketic calves showed th at
the principal alterations were confined to the skeleton. The liver, kid
neys and spleen were negative, but the gall bladders frequently con
tained as much as 500 cc. of a viscous (sometimes ropy) orange to
yellow colored bile. The gall bladders of normal calves contained
less than 100 cc. of am ber to olive-green colored bile.
Bile stained
ingesta occurred in the upper part of the small intestine in a few cases.
E n teritis was also occasionally seen. Excessive accumulations of syno
vial fluid in the joints were frequently observed in the more ricketic
calves. This m aterial was especially abundant in the larger joints and
varied from a thick, viscous fluid to a heavy, jelly-like substance.
The alterations in the skeletons may be more readily understood by
first studying the process of osteogenesis as discussed by Maximow
and Bloom (16) and by Leriche and Policard (17). An understanding
of the origin and importance of the epiphyseal or interm ediary cartilage
is especially desirable.
The Normal Costochondral Junction
The following detailed discussion of the normal costochondral junc
tion will serve as a w orkin g basis in interp retin g the histological pic
ture of ricketic bone. This description is based on material from five
norm al dairy calves which were between 161 and 317 days of age at
the time of slaughter.
F or convenience of description the ventral-epiphyseal end of the
rib, beginning at the union of the rib with the sternum and proceeding
vertebrallv, was divided into seven zones. The first five of these zones
are according to the divisions suggested by Dodds and Cameron (15)
for the epiphyseal cartilage (Fig. 3).
1.
Z o n e o f Reserve Cells—This area comprised the last several milli
m eters of the ventral end of the rib and usually term inated in a m oder
ately concave fashion where it joined the second zone. The tissue was
typical hyaline cartilage and was traversed in several places by vascular
bundles. "the diam eters of which were sometimes as g reat as 0.75 mm.
(Fig. 3). The num ber of vascular bundles was g re ater when the sec
ond^zone was approached. There were indications, as referred to below,
M IC H IG A N T E C H N IC A L B U L L E T IN NO. 150
th a t th e blood vessels in th e cartila g en o u s end of th e rib a n a s to m o s e d
w ith the vascular sy stem in th e diaphysis.
C artilage cell lacunae w e re irre g u la rly sca tte red , usually only p a r tly
filled by th e contained cells, and becam e m ore n u m e ro u s as th e second
zone w as approached. T h e y varied from oval o r cresce n t form s to m o re
spherical shapes which ch a rac te rized th e m a tu r e cells (F ig. 4). L a
cunae adjacen t to the la rg e r vascular bundles w e re fre q u e n tly flatten e d
w ith th e ir long 'axes tra n s v e rs e to th o se of th e bundles. T h e y o u n g ,
crescen t-sh aped cells w e re usually gro u p e d in pairs, except in th e v e n
tra l-p e rip h era l regio ns of Zone 1 w h e re as m an y as eig ht cells o c c u rre d
in one group. W h e n a r ra n g e d in isogenous pairs— and this see m e d
to be m ore freq uen t n ea r th e second zone— th e c resce n t shaped cells
w e re s itu ated w ith th e ir concave surfaces directly apposed to each
other. In this p a r t of Zone 1, th e in te rs titia l sub stan ce b e tw e e n is o g e n
ous pairs of cells w as som etim es invisible w h e n view ed in one plane, as
in a section, and tw o im m a tu re cells app e are d to be in one oval-sh ap ed
cavity (Fig. 4).
Individual cartilag e cells, w h e n n o t lost or sh ru n k e n , c o n fo rm ed in
shape to th e ir respective lacunae. T h e m a tu re r e s tin g cell, w h e n it
filled its lacuna, av e rag e d a b o u t 20 m icrons in lo n g e s t d iam eter. T h e
cytoplasm was reticular, m o d era tely basophilic, and con taine d several
vacuoles. T h e nucleus, w hich w as basophilic and reticular, w a s a b o u t
six m icrons in d iam eter and contained several c h ro m a tin g ran u le s.
C artilage cells w e re occasionally found which s u g g e s te d sta g e s in m i to
sis, while o th e r cartilag e cells appeared to be in v ariou s s ta g e s of
degeneration.
2.
Zone of Cell Proliferation — P ro ce ed in g fro m Zone 1 t o w a r d th e
diaphysis th e re w e re m an y pairs of isogenous cells a r r a n g e d p r e
dom inately a t first and la te r exclusively w ith th e ir lo n g axes p e r p e n
dicular to the long axis of th e bone. T his m a rk e d th e b e g in n in g of
Zone 2 (Fig. 3).
D a u g h te r cells, w h e n th ey o rig in ated in this area, ap p e are d to u n d e r
go im m ed iate division and g ave rise to fo u r cells. T his pro cess usually
continued and produced 8, 16, or m o re cells in one isogenous group.
F o r th e eig h th costochondral ju nction of th e d airy calf, th e n u m b e r
of such cells in one g ro up w hich appeared m o s t freq u en tly , a p p ro x i
m ated 32 cells— indicating t h a t each p a r e n t cell h ad divided five tim es.
This n u m b e r was variable, how ever, and it w a s difficult to d e te r m in e
ac cu rately w hen the cells w e re so closely apposed to each oth er. T h e y
frequ en tly failed even to ap p ro x im a te a m ultiple of tw o. T his m ay
have been caused by th e plane of sectioning, or possibly th e g ro u p w a s
not fully developed a t the tim e of tissue fixation or t h a t c e rta in cells
in such a g roup divided m o re actively th a n others.
Cells in th ese isogenous g rou ps continu ed to a r r a n g e th em selv es
d u rin g pro liferatio n w ith th e ir long axes perp en d icu lar to th e lo n g
axis of th e bone and piled one above th e o th e r so t h a t th e lo n g axis
of th e g ro u p w as parallel to th a t of the bone. T hus, a g ro u p or colum n
of cells arose from one p a r e n t cell. T h e s e colum ns ex ten d e d ac ro ss th e
bone w ith each colum n s e p a ra te d from its n e ig h b o r by h o m o g e n e o u s
in terstitia l substance. A dditional colum ns aro se and a r r a n g e d in m o re
or less ta n d e m fashion to form a ro w co n sistin g of g ro u p s o r colum ns
of cartilage cells. As n e w g roup s aro se th e older ones w e re seem in g ly
PATHOLOGY OF RICKETS IN DAIRY CALVES
7
pushed on to w a rd the diaphysis of the bone. Groups within row s w ere
sep arated by a thick mass of interstitial m atrix in contrast to the thin
tran sv e rse walls betw een cells within any one group.
C artilage cells in this zone were approxim ately of equal size and stage
of im m aturity. In th e axial p a rt of the bone this zone term in ated on
its v erte b ral side in a concave m anner as it m erged into the n ex t zone.
This concavity som etim es continued across the entire bone, in which
case Zone 2 had a uniform depth, but in m ost cases this zone becam e
deeper as it extended from the axial area tow ard th e peripheral p a r t
of the bone and usually attained m axim um depth a t 1.5 to 4.0 mm.
b en eath th e perichondrium. Zone 2 averaged about 600 microns in
depth in the axial region of th e bone and occasionally became as deep
as 1,300 m icrons in the peripheral p a r t of the rib.
3. Zone of Cell Growth—This zone was identified in two ways, first,
by th e g ra d atio n in size of cells, and second, by the gradual thinning
of th e interstitial m atrix (Fig. 3). G row th of the cells in this zone
w as apparently limited a t first to thickening in a plane parallel w ith
th e long axis of th e bone. L ater, w hen their thickness equaled their
width, these cells seemed to enlarge in all directions and ceased g r o w
in g w hen they attain ed an av erage diam eter of more than 30 microns.
G ro w th resulted in crowding and m utual pressure. T he cells as
sum ed m ulta n g u la r shapes, while th e m atrix betw een the cells w ithin
a g rou p gradually disappeared and the columnar a rran g e m e n t w ithin
th e isogenous groups became less perfect. There was lateral expansion
of these groups which involved a thinning of the interstitial m atrix
betw e en the rows.
M a tu re cells in this zone, except for their larger size and absence
of im m a tu re forms, w ere similar to those in Zone 1. In fixed section,
in about th e last 100 microns as Zone 4 was approached, th e cartilage
cells occupied a decreasing percentage of the areas in their lacunae.
The nuclei appeared shrunken, the cytoplasm increased in density, p a r
ticularly in the region of the endoplasm w here a condensation w a s
som etim es observed, definitely outlining and surrounding the nucleus.
S everal small vacuoles appeared in the cytoplasm. As these degenera
tive processes continued, the outline of the nucleus disappeared and
finally m ore than the usual num ber of em pty lacunae became visible.
T hus, while g ro w th was the predom inating feature of this zone, it
was nevertheless superseded in many, bu t not all, of the cells by w h a t
has been called hypertrophic degeneration.
4, Zone of Mature Cells—In the norm al rib, the m ost characteristic
fe atu re of this zone was calcification of the interstitial m atrix. This
calcified m atrix was later removed in its entirety. F o r this reason,
this is som etimes called the zone of tem p o ra ry or provisional calcifica
tion. This zone followed the outline of the previous area and showed
a continuation of the same degenerative changes which had not gone
to completion even in this zone (Figs. 3, 7, and 9).
In proceeding vertebrally th ro u g h this zone, coarse granules of a
basophilic substance w e re observed in the interstitial matrix. As these
granules increased in num ber, they produced a stippled effect. L a te r
they becam e obscured as they increased in num ber and became m o re
closely packed. This basophilic substance m arked the location of in
M ICH IGA N T E C H N IC A L B U L L E T IN NO. 150
organic salts. The salts w e re deposited in all of th e lon g itu d in al walls
between th e row s of cells and b etw e en th e g-roups of cells w ith in the
rows, b u t never betw e en the cells w ithin th e isogenous g ro u p s because
here th e in terstitia l m a trix appeared to be absent. T his point is i n t e n
tionally em phasized because of some debate over it in th e lite ra tu re .
Calcification w as o ften m ore com plete in th e m a rg in s t h a n in th e axial
areas of the longitudinal walls of ca rtila g e m a tr ix (Fig. 9).
This zone av e rag e d a b o u t 100 m icrons in depth. In a b o u t half th e
rases studied, it w as m uch deeper ju s t b e n e a th th e p eric h o n d riu m w h e re
the cartilag e row s w e re prolonged. H e re calcification, in exceptional
rases, extended for 1,000 m icrons b etw e en th e row s, b u t th e den sity
of m ineral deposition w as less and th e process w a s usually n o t co n
tinuous from the v e rte b ra l m a rg in of Zone 4. T h e re aso n fo r this
rxtended distribu tion of calcified m a trix is u n k no w n. P ossibly it adds
stren g th to the rib in the region w h e re it occurs.
5. Zone of Cartilage Removal— Zone 4 w a s b o rd e re d on its v e r te b ra l
m argin by a series of sac-like s tru c tu re s w hich fo rm e d th e zone of
cartilage rem oval (Fig. 7). Zone 5 w as 50-75 m icron s deep. T h e saclike s tru c tu re s w e re variable in size and ex tende d in th e d irectio n of
the long axis of the bone. T h e y w e re form ed by th e union of tw o or
more cartilag e cell lacunae follow ing th e d e stru c tio n of th e cell ca p
sules by the em bryonic m arro w . T h e m a rr o w elem en ts w e re slo w er in
rlestroying the calcified substance, so t h a t m a r r o w could be seen ad
vancing from the diaphysis to w a rd the v e n tra l end of th e rib in th e
form of n a r ro w to n g u es betw e en th e walls of calcified in te rs titia l su b
stance which acted as guides in the process. T h e sac-like s tru c tu re s
ivere bordered, th erefore, on e ith er side by calcified m atrix , while th e ir
vertebral ends w e re exposed to em bryonic m a rro w . T h e y m a y be
partially filled by d e g e n e ra tin g cartilage cells and by m a r r o w elem en ts
(Figs. 7 and 8).
6. Zone of Ossification—Exclusive of Zone 1, which varied in d ep th
depending upon the m an n er of dissecting th e rib from th e stern u m ,
the ventral-epiphyseal end of th e rib av e rag e d a b o u t 1,350 m icro ns
(w ith variations from 1.150 to 1,450 m icrons) in depth in th e axial r e
gion of the bone and ab ou t 2,100 m icrons (w ith varia tio n s from 1,150 to
1,350 m icrons) in th e peripheral region. As previously m entioned, vas:ular bundles extended from Zone 1 th ro u g h th e re m a in d e r of th e epi
physis and appeared to anasto m o se w ith th e vascu lar system in the
liaphysis.
Zone 6 co n stituted the b eg in ning of the diaphysis. T h e zone usually
dad a convex outline on its v en tra l m a rg in (Fig. 11). E m b ry o n ic m a r
row w as observed advan cin g from th e diaphysis t o w a r d th e v e n tra l
end of th e bone. In advance of this, ery th ro c y te s , fibrin and so m etim e s
serous m aterial, and occasional endothelial cells w e re fre q u e n tly ob
served, closely followed by larg e n u m b e rs of em bryon ic connective
tissue cells (Fig. 7). D e g e n e ra tin g ca rtila g e cells freq u e n tly appe are d
to be revived at this point, because of a new source of n u tritiv e m a t e
rial from the diaphysis.
Calcified tissues resisted rem oval by th e m a rro w . A b o u t 50 m icron s
vertebral to Zone 5, cells w e re occasionally seen sim ilar in size to th e
primitive w a n d e rin g cells of th e em bryonic connective tissue lining th e
PATHOLOGY OF PICKETS IN DAIRY CALVES
persistin g bars of calcified m atrix (Figs. 7 and 13). These were osteo
blasts, the cells associated w ith the production of acidophilic layers of
osteoid tissue occasionally observed on these trabeculae. Osteoblasts
averag ed about 16 microns in diam eter and were mononuclear. The
nucleus m easured about seven microns in diam eter and contained one
or m ore large nucleoli eccentrically placed near a pale attraction sphere.
I h e cytoplasm was reticular, intensely basic, and contained small vac
uoles. T h e calcified bars of m atrix appeared to serve at least three
purposes. T hey stren g th en e d the area in which they occurred, served
as guides for the advancing m arro w in cartilage removal, and formed
bases for the deposition of osteoid tissue. M ost of these bars p e r
sisted into the diaphysis to about 700 microns vertebral from Zone 5
and then m any of them w ere gradually removed by osteoclasts. O steo
clasts appeared to function specifically in the removal of calcified
cartilage and osseous tissue, and in this w ay aided in the advancement
of the em bryonic m arro w and at the same time assisted in the internal
reconstruction of bone. Osteoclasts w ere m ost noticeable on the v erte
bral ends of persisting bars of calcified m atrix w here they caused t e r
minal erosion of the bars (Figs. 5, 15, and 16). These cells, however,
were also observed about as far ventrally in the rib as w ere osteoblasts.
Osteoclasts w ere variable in size and contained from two to 25 or more
nuclei. Each nucleus was about five microns in diam eter and contained
several coarse granules. The cytoplasm was vacuolated, took a less
basic stain th an th a t of the osteoblast, and frequently showed pseudopodal projections. The means by which osteoclasts erode calcified
m atrix and bone have not been included in this study. W h e th e r the
process of erosion is one of osteolysis or of phagocytosis is a debated
question.
P ers is tin g trabeculae of calcified tissue eventually lost their cal
cified cartilage cores by chondrolysis, and the rem aining trabeculae
of bone became thicker fu rth e r in the diaphysis by peripheral additions
of osteoid tissue. The presence of n arrow margins of osteoid tissue
at any one time indicate early ossification in the normal rib. The line
of dem arcatio n betw een osseous and osteoid tissues was a gradual one
m arked by an increase in the num ber of basophilic granules so th at
the stippled effect gradually became more dense in passing from osteoid
to osseous tissue. These granules sometimes appeared to be deposited
in striations (Fig. 5). Osteoid tissue was always accompanied by osteo
blasts but the converse was not always true. Frequently m any osteo
blasts, instead of a single layer, w ere seen bordering trabeculae, and
whole clumps of these cells occurred in crotch-like areas where two
trabeculae had united (Fig. 14). Individual osteoblasts were commonly
noticed in the process of .being surrounded by osteoid tissue (Fig. 13).
A fte r incorporation in the bone these osteoblasts gradually shrank
and assum ed the flattened character of osteocytes (bone cells). In
stained preparatio ns the osseous m aterial in the trabeculae exhibited
various shades of basophilia. This fact has suggested the possibility
t h a t different degrees of calcification w ere represented in the bone.
D ecrease in the num ber of persisting trabeculae was accompanied
by larg e r m arro w spaces betw een the trabeculae. A t about 15 mm.
v erteb ral from Zone 5 the embryonic m arrow had disappeared and
hemopoietic foci, along with vacuoles (probably of fatty origin) a p
LO
M ICH IGA N T E C H N IC A L B U L L E T IN NO. 150
peared and ev entually com pletely m a sk e d th e loose connective tissue
f u rth e r in th e diaphysis. H em o c y to b la sts, ery th ro b la s ts, a n d n e u t r o
phil m y elo cytes co n s titu te d th e m a jo rity of th e cells. M a n y sm all foci
of lym phoid cells w e re s c a tte re d in th e red m a rro w . H e r e o s teo b lasts
and osteoid tissue, as well as osteoclasts, w e re lim ited la rg e ly to t h e
p erip heral re gio ns of the diaphysis on tra b e c u la e closely b e n e a th th e
periosteum .
7.
Zone of Compact Substance—T w o questions a re considered u n d e r
this zone which has been called, for w a n t of a b e t t e r nam e, th e zone of
com pact substance. F irs t, w h a t accounts for th e n a r r o w i n g of th e rib
bone in p assing fro m th e ventral-ep ip h y sea l end to w a r d th e v e r te b ra l
end? A nd second, w h a t so rt of co v ering does th e rib p ossess to sep
a r a t e it from su rro u n d in g tissu es?
T h e cartilageno us end of th e rib w as covered by a m o d e ra te ly dense
connective tissue (the p erich ond rium ) w hich v arie d fro m 150 to 500
m icrons in thickness. A t th e ju nctio n of Zones 1 and 2 th e p e ric h o n
drium usually exten ded as a less dense connective tissu e in w a rd to a
distance som etim es as g r e a t as 800 m icrons. W h e n view ed in one plane,
as in a section, it g ave th e im pression of a cone w ith th e ap e x direc ted
to w a rd th e c e n te r of th e bone (Fig. 11). Blood vessels fre q u e n tly
covered the crest as well as th e lateral b o rd e rs of this area. T h e p e r i
chondrium extended over th e zones of cell p ro life ra tio n and cell g r o w t h
as a denser connective tissue of 200-300 m icron s in th ickness. B e
ginning w ith Zone 5 it continued as th e perio steu m . R a re ly , a n a r r o w
strip w ithin the perich on driu m over Zone 4 ap p e a re d to be calcified.
A t 1.5 mm. v erte b ral from Zone 5 th e p e rio s te u m th ick en e d by e x te n d
ing as a looser connective tissu e in w ard b e tw e e n th e p e r s is tin g t r a b e
culae to a distance of 1.5-2.0 mm. and th e n g ra d u a lly d ecrea sed to 150
m icrons o r less a t 25 mm. v e rte b ra l fro m Zone 5 w h e re it only ex ten d e d
in b etw e en a few of th e p erip heral trabeculae. T h is d ec rease in th ic k
ness of th e p erio steu m o ccu rred a t a p o in t of in cre a s in g size of p e r i
ph eral trabeculae, which la te r g re w th ic k e r and assum ed th e c h a ra c te r
of a m o re com pact m a teria l th a n the tra b e c u la e in th e m a r r o w cavity.
Gross m ea su re m e n ts of m id -fro n ta l sections indicate t h a t th e rib
was w idest s o m ew h e re n e a r th e jun ction of Zones 5 and 6 w h e re it
varied from 20 to 30 mm. in w id th in n o rm a l calves of th e ag es u n d er
consideration. T h ro u g h a process of n a rro w in g , so m etim e s re f e r r e d to
as tubulation, the rib g ra d u ally decreased to a m in im u m w id th of
4-8 mm. a t a b o u t 50 mm. v e rte b ra l from Zone 5. T his tu b u la tio n o c
cu rred by term in a l erosion of th e p e rs is tin g tra b e c u la e of b o n e w hich
occur in the e x tre m e p eriph eral areas and w hich ex ten d parallel to th e
long axis of th e bone and th e re fo re join th e p e rio s te u m a t an oblique
angle. Evidence of this erosion was t h a t one .or m ore o s te o cla sts w e re
observed adjac en t to th e v e rte b ra l ends of th e se tra b e c u la e (F ig. 6).
This erosion, w hich in all probability w e a k e n e d the bone, w a s c o m p e n
sated by th e extension of th e p e rio steu m into th e in te rio r of th e bone
in the m a n n e r described above.
The Costochondral Junction in Rickets
This description is based on m ate ria l fro m 11 calves w h ich w e re
suffering from vitam in D deficiency and w hich w ere, except fo r one
anim al which died from com plications of ric k e ts a t 520 days, b e tw e e n
PATHOLOGY OF RICKETS IN DAIRY CALVES
11
151 and 330 days of age at the time of slaughter. The duration of the
ricketic condition varied from 38 to 239 days. The cases are about
equally divided betw een (a) low blood plasma calcium rickets and (b)
low blood plasm a calcium-low inorganic phosphorus rickets. No cases
of low plasm a inorganic phosphorus rickets, uncomplicated by low
calcium, are included in this study.
The alterations which w ere observed in the ricketic bones will be
considered according to the zones in the norm al rib. The histological
sections will be a rran g e d in sequence of events as they occurred du r
ing the developm ent of the ricketic rib from the normal rib.
1. Zone of Resting Cartilage—This zone usually term inated in a
m od erately concave m ann er on its vertebral side similar to th a t in the
n orm al rib. In prolonged rickets it varied to the ex ten t of m eeting
Zone 2 in a convex m anner. In the cases of prolonged rickets there
was an increase in the num ber of oval-shaped cavities apparently formed
by the union of two lacunae, irregularly arranged adjacent to Zone 2
and similar to those noted in the normal bone (Figs. 17 and 18). This
alteratio n was m ost pronounced in severe rickets when it involved as
m uch as th e last tw o mm. adjacent to Zone 2. In rickets of shorter
duration, these enlarged cavities appeared only in isolated patches
adjacent to Zone 2. In such areas the m atrix betw een isogenous pairs
of cells was reduced to a thin network. The m ature cartilage cells were
few er in num ber and the impression was obtained th at the decrease
in the ra te of g ro w th had resulted in the accumulation of im m ature
cartilage cells.
2. Zone of Cell Proliferation—At the junction w ith Zone 1, the o u t
line of this area a t first became m oderately undulated. As the severity
of rickets increased, this general outline became more convex on its
ven tra l m argin in th e axial region of the bone, w hereas in prolonged
disease it even extended into the resting cartilage in the axial p a rt of
the rib in a wredge-shape manner. The apex of the wedge was som e
times r a th e r broad, while the sides extended out from the base tow ard
the peripheral p a rt of the bone w here they either term inated in the
usual m an n er or curved ventrally for some distance just beneath the
perichondrium . In addition to the altered outline, Zone 2 revealed an
irregular decrease in depth in rickets and was as shallow as 100 microns
in certain places while of norm al dimensions in others. There was some
evidence t h a t the average num ber of cells per isogenous group was
reduced in rickets (Fig. 17).
3. Zone of Cell Growth—This area followed the outline of the
previous zone in the same bone. I t w as unaffected in depth, except
in ra re instances when it was partially obliterated in places on its
verte b ral side by sinusoidal cavities containing embryonic m arrow
(Fig. 17).
4. Zone of Mature Cells—W h ereas the changes thus far noted in the
cartilagcnous end of the rib w ere not always conspicuous, especially
in mild rickets of short duration, the alterations in Zone 4 were more
pronounced and w ere therefore nun e consistently obseivcd. th e s e
chaimes m ay be followed with considerable satisfaction when the se
quence of events is traced in the costochondral junctions of the
12
M ICH IGA N T E C H N IC A L B U L L E T IN NO. 150
ricketic ribs w h e n th e y are em pirically a r ra n g e d in ord e r of in c re a s
ing severity of rickets.
R e ta rd e d provisional calcification w as th e a lte ra tio n first observed
(Fig. 9). H ere, Zone 4 was of n orm al dep th b u t th e a r e a of provisional
calcification extended irre g u la rly into th e m atrix , fro m th e v e r te b ra l
side of this zone, and av e rag e d less th a n 50 m icrons in depth , while
in places it m easu red zero w h e n it failed to ex ten d v e n tra lly bey o n d
th e fifth zone. As th e sev erity of rick e ts increased, o nly occasional
longitudinal walls of cartilag e m a tr ix b etw e en the ro w s of m a t u r e cells
w e re calcified. In m ore advanced rick e ts calcification w a s en tire ly
ab se n t in this are a and frequ en tly only involved a rela tiv e ly small
am o u n t of the m a trix in Zone 5 ad jac en t to Zone 4.
L ac k of calcification in Zone 4 w as soon followed by le n g th e n in g of
the ro w s of m a tu re cartilage cells. A t first, c a rtila g e ton g u es, each co n
sisting of a g ro u p of cell row s and th e ir in te rv e n in g m atrix , p ro je c te d
v erte b rally for a sh o rt distance into Zone 5 and g ave an u n d u la tin g
outline of the v erte b ral m a rg in of Zone 4 (Fig. 10). L e n g t h e n in g of
th e row s becam e m o re generalized in m ore advanced ric k e ts and this
zone becam e deeper across the entire w idth of th e bone (F ig. 19). T h e
n u m b er of m a tu re cells in row s w as g r e a t e r in p rolo n g ed rick e ts w h e n
this zone reached a depth as g r e a t as 800 m icrons in th e axial region
of the bone and as g r e a t as six mm. in th e m o re p erip h e ral p a r t of th e
bone (Fig. 20). T h e m a trix b etw e en these le n g th en e d colum ns of cells,
while hom ogeneous in appearance and free of g ra n u le s of calcified su b
stance, tend ed to w a rd a deeper basic stain th a n t h a t in Zone 3. T h e
m a tu re cartilage cells in this zone in prolo nged rick e ts av e ra g e d less
t h a n 20 m icrons in diam eter. This decrease in cell size w a s prob ab ly
a result of pressure, e x a g g e ra te d by d y stro phic conditions a risin g from
the increase in size of th e v e g e ta tiv e c a rtila g e mass. S everal stag es
of d e g e n era tio n w ere observed and cells w e re dislodged from m an y
lacunae. C artilag e cell re juv en ation , how ever, w a s co m m o n ad jac en t
to Zone 5 (Fig. 22).
Zone 4 followed on its v en tra l side th e o utline of Zone 3. On its
v erte b ral m a rg in tw o factors helped to p ro du ce an e x tre m e ly i r r e g
ular outline a t the jun ction of Zones 4 and 5. One of th ese w a s th e
cro w ding of th e v e rte b ra l ends of the lo n gitu din al row s of cells which
caused w hole to n g u e s of cartilag e to bend so t h a t the ro w s of cells in
these to ng ues no lon ger extended parallel to th e lon g axis o f th e bone.
This bu ckling of cell row s w as favored by th e lack of provisio nal cal
cification (Figs. 21 and 22). T he second factor, w hich o p e ra te d to p r o
duce an irre g u la r outline at the junction of Zones 4 and 5, w as the
uneven ad vancem en t of the em bryonic m a r r o w from th e diaphysis
(Fig. 12).
Zones 2, 3, and 4 in the ricketic bone m ay c o n trib u te as m uch as
tw o mm. in the axial area, and as m uch as seven mm. in th e p erip heral
region, to the leng th of the rib. This thick ene d c a rtila g e a t the v en tra l
end of the rib was trav e rsed , as in th e no rm al bone, by v asc u la r bundles
w hich appeared to a n a sto m o se w ith vessels in th e diaphysis.
5.
Zone of Cartilage Removal—T his zone pro b a b ly sh ow ed m o re
pronounced alteratio n s th a n any o th e r p a r t of th e bone in rick e ts (Figs.
—1 and 23). In eai 1) lickcts, cuililage icm o v al con tin u ed at a relativ ely
more rapid ra te than provisional calcification so th a t ca rtila g e removal,
PATHOLOGY OF RICKETS IN DAIRY CALVES
13
while occurring in the usual manner, advanced ventrally to a position
ab re a st with, but never ventral to, the line of provisional calcification.
A pparently the embryonic m arrow does not normally remove cartilage
w hen the m atrix betw een the cells is uncalcified, which accounted for
the accum ulation of m ature cartilage cells in Zone 4.
In m ore severe rickets there was a failure of calcification in Zone 4
and also in m ost of the m atrix in Zone 5. The cessation of normal
cartilage rem oval halted growth-involving ossification. Such cessation
in grow th, however, was in some w ay compensated, as illustrated by
h eig h t-a t-w ith e rs m easurem ents of severely ricketic calves which con
tinued to gain in heig h t although at a reduced rate, even when body
w eight was stationai'y or decreasing. The rib shared in this skeletal
g r o w th in rickets. At least partial compensation for lack of calcifica
tion occurred as fo llo w s : In rickets of long duration the rows of m a
tu re cartilage cells lengthened appreciably, which sometimes am ounted
to as much as six mm., and a new m ethod of cartilage removal was
apparent. The embryonic m arrow which advanced ventrally from the
diaphysis was still the active agent of cartilage removal, b u t instead
of p ro gressing evenly in n a rro w tongues, it was unguided by cal
cified walls of m atrix and advanced unevenly and in a m ore or less
oblique m an n er in broad tongues into Zone 4. This advancem ent de
stroyed wide areas of cartilage, including cells and m atrix—both of
which appeared to be destroyed with equal ease and rapidity (Figs.
12 and 20). E xtrem e ly large and irregularly shaped sac-like structures
occurred adiacent to Zone 4. These contained in addition to the ele
m ents seen in norm al sac-like areas, num erous erythroblasts and hemocytoblasts enmeshed in loose connective tissue (Figs. 9, 12, and 20).
Osteoblasts w ere observed in advance of osteoclasts, generally, although
the reverse som etimes occurred (Fig. 9). The cartilage m atrix which
partially surrounded the sac-like areas assumed an acidophilic reaction.
The m a rro w invariably advanced fu rth e r ventrallv, sometimes by as
m uch as four mm., in the axial region of the bone (Fig. 12). The g reat
unevenness of the m a rro w advancement gave the appearance of w hat
has been referred to as “irregular thickening of the cartilage” and was
occasionally exagg erated by actual splitting of the vertebral p art of
Zone 5 (Fig. 21). Irre g u la r advancem ent of m arrow led in some places
to sac-like areas of embryonic bone m arrow which appeared, in one
plane as in a section, to be isolated in the zone of m ature cells (Fig. 20),
and in o th er places it led to isolated masses of cartilage in the diaphysis.
J u s t v erteb ral to the zone of sac-like areas, th ere occurred patches
of connective tissue in which the central parts had taken a deeper
acidophilic stain identical with th a t of osteoid tissue (Figs. 23 and 24).
Im bedded in such tissue w ere one or several cells th a t resembled osteo
blasts, su ggestin g th a t certain of the local hemocytoblasts had differ
entiated into osteoblasts which had, in turn, elaborated osteoid tissue.
These new patches of osteoid tissue served as bases for additional
osteoblast activity. Occasionally, isolated masses of cartilage with
acidophilic m a trix appeared in which the cells had not only been r e
juvenated but had also been transform ed to osteoblast-like cells, sug
g e stin g th e possibility th a t cartilage can be changed en masse into
osteoid tissue. Isogenous pairs of cells were observed in the larger
masses of cartilage along with degenerating cartilage cells. The net
14
M ICH IGA N T E C H N IC A L B U L L E T IN NO. ISO
re su lt of all of this cell activ ity w a s a zone co n sistin g larg e ly of osteoid
tissue. B ecause of th e m a n n e r in w hich th e o steob lasts a p p e a re d to
develop, th e ch a ra c te ris tic a r r a n g e m e n t of th e s e cells b o r d e r in g on
trabeculae, as revealed in n o rm a l ribs, w a s inconspicuous in spite of
the la rg e a m o u n t of osteoid tissu e t h a t w as p re s e n t (F ig. 23). T h e
osteoblasts in this re g ion of th e bone have been called “O s te o id o b la s ts ”
by some w o rk e rs on th e basis t h a t th ese cells prod uce tissu e w hich
does n o t ossify and th ere fo re , th e cells were* r e g a rd e d as d e g e n e ra te
form s of osteoblasts. Such a distinction is unjustified in th is s tu d y
because th ese cells ap peared to be identical in th e i r a d u lt fo rm w ith
the osteoblasts in n orm al ribs. I t does n o t seem illogical, th e re fo re , to
ascribe th e failure of ossification in ric k e ts to an y one of seve ral causes
— such as a deficiency in v itam in D — r a th e r t h a n to an in h e re n t d efect
in th e osteoblasts. O steoclasts w e re few in n u m b e r in this region. T h e y
seem ed to have no specific function in this a r e a b u t ap p e are d because
of th e ir p ro x im ity to th e diaphysis.
T h e osteoid tissue varied considerably in d ep th in any one bone b e
cause of irre g u la r m a rr o w adv an cem ent, b u t it m e a s u re d as m uch as
six mm. in dep th in m ore p rolo ng ed rickets. In m ilder rick e ts th e n o r
m al and abno rm al types of cartila g e rem o v a l w e re ob served in th e sam e
bone (F ig . 19).
T h e v e rte b ra l outline of Zone 5 w as stro n g ly concave w h e re it m e rg e d
into Zone 6. This tra n s itio n in zones w a s a g ra d u a l one in w hich b a s o
philic g ra n ules appeared in isolated areas and s u rro u n d e d individual
cells in osteoid tissue. Such cells w e re conspicuous b ecause of th e
c h a ra c te ristic halo abo u t each one (Fig. 23). T h e halo bec am e n a r r o w e r
w ith increase in th e n u m b er of g ra n u le s and in creased ossification n e a r e r
the diaphysis. T h e re also appeared small tra b e c u la e of bone, su rro u n d e d
on all sides by b road layers of osteoid tissue. O steo b lasts soon b o rd e re d
these trab e cu lae in th e usual m a n n e r as Zone 6 w as approached.
6. Zone of Ossification—R e o rg a n iz a tio n of m a te ria l b e tw e e n th e
patches of osteoid tissue occurred soon a f te r th e first signs of ossifica
tion w e re observed, and it w as h ere t h a t Zone 6 b e g a n (Fig. 23). P r o
ceeding vertebrally, th e spaces b etw e en tra b e c u la e enlarge, and th e
em bryonic m a rr o w w as soon replaced by loose connective tissue. S ev
eral m illim eters v e rte b ra l to Zone 5, h em o po ietic elem ents' and vacuoles
began to m ask the loose connective tissue in th e m a r r o w spaces. N e u
trophil m yelocytes w e re as n u m ero u s h ere as in th e n o rm a l re d 'm arrow .
In th e e x tre m e v e n tra l are a of Zone 6 th e tra b e c u la e w e re s h o r t an d
stubby and w e re b o rd e red by b ro a d osteoid tissu e m argins. T h e y also
revealed fre q u e n t tra n s v e rs e a n a sto m o ses (Fig. 20). M o re v e rte b ra lly
in th e diaphysis th ese trab e cu lae a s su m ed a m o re n o rm a l shape and
distribution as the osteoid tissue m arg in s g ra d u a lly b ecam e n a r ro w e r .
In the red m a rr o w region, trab e cu lae con tin ued to sh o w ab n o rm a lly
broad m arg in s of osteoid tissue, w hich som etim e s ap p e a re d to be
more sharply d e m arcate d from osseous tissue in rick e ts t h a n in n o rm a l
bone. Chondrolysis w as an u n im p o rta n t g r o w t h f e a tu re h e re b ecause
the ap pearance of calcified c a rtila g e cores w a s in v erse ly p ro p o rtio n a l
to th e sev e rity of rickets.
7. Zone of Compact Substance— A t th e ju n ctio n of Zones 1 and 2
the perichondrium was thickened and som etim es ex ten d e d in w a rd as
PATHOLOGY OF RICKETS IN DAIRY CALVES
IS
far as 2.5 mm. I t continued in a m oderately thickened form over Zones
2, 3, and 4 and in this location trabeculae of bone, m ediary betw een the
ex ternal and internal surfaces, were usually seen. These trabeculae
appeared to have the same origin as those in the v ertebral p a r t of
Zone 5. T he reo rgan ization of the m aterial betw een trabeculae oc
curred in a similar m an ner and resulted in the form ation of m arro w
spaces which continued fu rth e r vertebrally as a p a rt of th e m arrow
cavity. T he perichondrium continued as periosteum of about normal
thickness.
Tubulation appeared to occur in the norm al manner. The beading
or enlargem ent of the ventral end of the rib, which was commonly
observed in gross a t the tim e of slaughter, was an inconspicuous fea
tu re of the bone w hen studied w ith the microscope. Gross m easu re
m ents of m id-frontal sections showed th a t the diam eter of ricketic ribs
varied from 20 to 35 mm. w hereas the range of norm al bones was
from 20 to 30 mm. The average diam eter was about six mm. g re a te r
in the ricketic bone. Microscopic observations showed t h a t the g r e a t
est diam eter was th ro u g h the area of osteoid tissue in Zone 5, in the
region ju st v erteb ral to th e cartilagenous end of the rib. An excess
of osteoid tissue appeared to be the cause of this increase in size of
the rib end. This enlarg em en t was more apparent than real, however,
because the cu rvature of the medial border of the ventral end of the
rib was g re a te r in rickets Figs. 25, 26, 27, and 28). The curvature of
the lateral border increased in the same direction. This change in c u r
v a tu re was g re a te s t at about 20 mm. vertebral to Zone 1 and could be
the result of a w eakened rib responding to norm al physiological de
mands such as respiration a n d /o r support of the diaphragm. This in
w a rd curving of th e ven tral end of the rib exagg erated any enlarge
m e n t t h a t did occur, especially when viewed from the medial border
of the bone.
The Costochondral Junction in Healing Rickets
Calf C-224 was the only animal included in this phase of the study.
This calf developed low plasma calcium rickets at 110 days of age.
T he daily am oun t of whole milk fed was increased at a later date,
and a t 140 days of age the plasma calcium began to increase and was
nearly norm al when th e animal was killed at 163 days of age (Table 6).
This case is of in terest because it appeared to resemble the epiphyseal
type of healing rickets usually observed in the rat. This was best illus
tr a te d in the ro en tg e n o g ram which showed at about six mm. vertebral
to the cartilagenous end of the bone, a n arro w region extending half
w ay across the bone from the medial side and which also showed a lack
of ossification (Fig. 26). Histologically, a more or less normal costo
chondral junction was observed except in the n arro w uncaldfied area
in Zone 6, w here the trabeculae w ere small and scattered in an area
consisting m ostly of connective tissue.
Roentgenographic Appearance of Normal and Ricketic Ribs
R o en tg en o g ra m s of norm al ribs showed a sharp line of demarcation
a t the junction of the diaphysis w ith the cartilagenous end of the bone
(Figs. 25, 26, and 28). The diaphysis usually ended in a predominately
16
MICHIGAN T E C H N IC A L B U L L E T IN NO. 150
convex m an n er w h e re it joined the cartilage. T ra b ecu lae of osseous
tissue w e re conspicuous and indicated a d istrib u tio n of in o rg a n ic salts
grossly identical w ith t h a t described above in th e histological s tu d y of
the n orm al rib. T h e v en tral-ep ip hyseal end of th e rib curv ed in a
medial direction. T h e walls of th e diaphysis becam e th ic k e r in p r o
ceeding v erte b rally fro m th e costochondral junction.
Osteoid tissue has th e sam e hom ogen eou s ap p e ara n ce as c a rtila g e in
th e ro e n tg e n o g ra m so t h a t ce rtain definite a b n o rm a lities w e re visible
in rickets. T h e line of ju nction of c a rtila g e and diaphysis w a s i r r e g
ular and indefinite, and consisted of an area, v a r y in g in depth, in w hich
incom plete calcification of tissues had occurred. T h e so-called “ cup
ping,” or d evelopm ent of a concave outline of th e v e n tra l end of th e
diaphysis, w as n o t a cha rac te ristic fe a tu re in th e rick e tic calf rib in
the specimens studied. In m an y cases, th e end of th e diaphysis b ec am e
m ore convex in rickets, due to the ad v a n cem en t of em bryonic m a r r o w
in the axial region of th e bone.
O th e r t h a n an increase in medial c u rv a tu re of the rib, th e ro e n tg e n o grap hic evidences of rick e ts in th e specimens studied w e re confined
larg ely to the are a of excess osteoid tissue which s e p a ra te d th e v e n tra l
end of th e diaphysis from the ca rtila gen ous end of th e bone. T h e im
po rtan c e of this localization of ricketic chang es is discussed below.
Growth As a Modifying Factor in Rickets
T h e effect of g r o w th upon rickets is illu stra te d w h e n calves C-237
and C-238 are com pared (Tables 8 and 9, Figs. 19 and 20). T h e se a n i
mals received ab ou t th e same am o u n t of v itam in D in th e fo rm of w hole
milk, and bo th developed low plasm a calcium rick ets by 90 days of age.
T h ese calves w e re s la u g h tere d a t ab ou t th e sam e age. T h e r a te of g ain
in body w e ig h t w as m uch slow er for C-237. H isto log ical studies show
very definitely t h a t th e ricketic a lteratio n s in th e rib w e re m u ch less
extensive in the slow er g ro w in g calf.
A com parison of calves C-170 and C-232 m ay be used to illu s tra te
the influence of ag e in rickets (Tables 5 and 7, Figs. 10 and 29). C-232
developed low plasm a calcium rickets by 100 days of age, exhibited
low in organic ph osp ho rus values at 140 days, and w as s la u g h te re d at
161 days of age. C-170 developed low p lasm a calcium ric k e ts by 170
days of age, th e in org anic p hosphorus fluctuated c o n s id e ra b ly ' a t this
tim e b u t w as n ot definitely s u bno rm al until a f te r 280 days of age. This
anim al w as s la u g h tere d a t 330 days of age. B o th calves gain ed subnorm ally in body w eight. K eepin g in mind the ag e difference an d th e
fact t h a t the older calf had rickets for 160 days, w hile th e y o u n g e r
calf had ab out th e sam e severity of rick e ts for only 61 days, as ju d g ed
by the blood plasm a analyses, it is significant t h a t in ro e n tg e n o graphical as well as in histological studies, th e y o u n g e r calf (C-232)
m anifested th e m ore ex tensive ricketic alteratio ns, alth o u g h it s u f
fered from the disease fo r a m uch s h o rte r time.
Complications in Rickets
Calf C-188 (T able 5) developed rick e ts a t 380 days of ag e and in
addition to the usual sy m p tom s the anim al show ed muscle a t r o p h y and
e x tre m e em aciation. C-188 died at 520 days of age, follow ing a c o m a
PATHOLOGY OF RICKETS IN DAIRY CALVES
17
tose condition and failure to respond to subcutaneous injections of
viosterol and intravenous injections of calcium gluconate and m a g
nesium sulphate solutions. I t lost more than a pound of body w eight
daily during the last 100 days of life.
On post m o rtem examination, th e m arked pitting and erosion of the
artic u la tin g surfaces w ere m ost conspicuous, particularly at the distal
end of the fem ur and the proximal end of the hum erus (Fig. 33). This
condition existed in th e peripheral as well as the central p a rts of the
articular areas. W h e re the cartilage surface was not destroyed, it was
v ery loose and easily removed by the lingers. Bone of a porous-plate
appearance w as exposed. Spongy pads of protein-like material covered
these areas. R o entg en o g ra m s of thin sections of bone removed from
such areas showed an irregular rarefaction of the bone beneath the
eroded or pitted surfaces (Fig. 32). Histological studies indicate th a t
fibrous connective tissue occupied these rarefied, calcium-free areas.
In the ven tral end of the rib a few faint indications of arrestedg r o w th lines, m ark in g areas of increased num bers of transverse trab e c
ulae, w ere observed in the ro e n tgen ogram vertebral to the cartilagen
ous end of the rib (Fig. 25). Histologically, provisional calcification
w as som ew hat re ta rd ed and incomplete. A shallow area of calciumfree osteoid tissue was vertebral to Zone 4. Following this area were
sh o rt trabeculae of bone with broad margins of osteoid tissue and more
th a n th e norm al num ber of transverse anastomoses. T h e lack of ex
tensive changes in this ricketic rib was attribu ted partly to the age
of th e animal, and p artly to the severity of the disease and its halting
effect upon grow th.
,
Type of Rickets and Histological Alterations
W h en observed in prepared sections, the histological alterations in
th e rib appear to be identical in low plasma calcium rickets and in
rickets associated with low calcium and low inorganic phosphorus.
DISCUSSION
R ickets in dairy calves suffering from a deficiency of vitam in D was
ch aracterized by changes in the bones (Figs. 1-34), which were ap
p aren tly accompanied by decreased concentrations of the plasma cal
cium a n d / o r inorganic phosphorus (Tables 2-9). Other changes, more
inconsistently found, w ere the accumulation of bile in the gall bladders
and the presence of bile stained ingesta in the upper region of the
small intestine. The costochondral junction at the ventral end of the
rib w as selected as the best and most convenient index to the degree
of rickets in calves (Figs. 1 and 12).
W h e n com pared w ith the norm al cases, the ricketic costochondral
junctions w ere characterized fundamentally by retarded and incom
plete provisional calcification of the cartilage matrix, which in turn
caused te m p o ra ry failure of cartilage removal by the embryonic m a r
ro w (Figs. 9 and 10). M ature cartilage cells then accumulated up to a
point w here a ricketic type of cartilage removal was initiated by the
em bryonic m arrow . It was this la tte r process which caused the most
conspicuous alteratio ns in the rib (Figs. 12, 19, 20, 21, and 29).
8
M ICH IG A N T E C H N IC A L B U L L E T IN NO. 150
I f th e th e o ry is accepted t h a t r e ta rd e d provisional calcification is
the fu n d a m e n tal ch a n g e in th e bone in ric k e ts — and t h e r e is e v e iy
reason to believe t h a t such is th e case in ric k e ts caused b y a v ita m in D
deficiency— th e n th e histological alte ra tio n s in th e rick e tic b o nes m ay
be explained in w h a t app e ars to be a v e ry s a tis fa c to ry m a n n e r. Thus,
in th e absence of th e lo ngitudinal trab e cu lae of calcified c a rtila g e m a
trix to act as guides, the em bryonic m a rr o w in th e rick e tic ty p e of c a r
tilage rem oval advanced irre g u la rly and obliquely in b ro a d to n g u e s
which caused an uneven decrease in d ep th of th e zone of m a t u r e cells
(Fig. 12). T h e irre g u la r m a n n e r of m a rr o w ad v a n c e m e n t led to sac-like
areas which appeared, in one plane as in a section, to be iso lated in th e
zone of m a tu re cells. In o th e r instances it re s u lted in w h a t ap p e are d to
be isolated m asses of cartilage v e r te b ra l to th e zone of m a t u r e cells.
V ertebrally, in th e region adjacent to th e la rg e sac-like areas, w e re seen
patches of connective tissue which occupied m o s t of t h a t p a r t of th e rib
(Fig. 23), In th ese p atches h em ocy to b lasts ap pe are d to ch a n g e to o s t e o
blasts which produced areas of osteoid tissue in th e p atch e s of fibrous
connective tissue (Fig. 24). E v e n tu a lly m o s t of this zone w a s occupied
by osteoid tissue, th e accum ulation of w hich w as pro b a b ly of a co m
p e n s a to ry n a tu re in th e absence of bone fo rm a tio n . T h us, in re a lity
ca rtila g e rem oval in rick e ts w as fu n d a m e n tally th e s am e as t h a t in th e
no rm al bone, b u t a different ap pearan ce w as prod uced b ecause of th e
lack of provisional calcification. C hanges w e re less exten siv e f u r th e r
v erte b rally in the diaphysis as th e bone ap p ro ach ed a m o re n o rm a l
appearance.
T h e beading or swelling com m only o bserved in th e v e n tra l ends of
ribs from rick etic calves was n o t a conspicuous chang e w h e n view ed
in th e microscope. T h e re was, h ow ever, a m o d e ra te sw ellin g of th e
v e n tra l end of th e bone, th e origin of which ap p e are d to be due to th e
accum ulation of osteoid tissue. This e n la rg e m e n t w as e x a g g e r a te d by
the increased cu rv a tu re of the medial bo rd e r of th e rib (F igs. 25, 26,
and 28).
R ickets affected the g ro w in g ends of bones (F igs. 25, 26, and 28).
The alte ra tio n s w e re less severe in th e slow er g r o w i n g calves (Figs.
19 and 20) and also in th e older anim als w h e n th e r a t e of g r o w t h had
decreased (Figs. 10 and 29). U n d e r the conditions of this stu d y g r o w th
occurred a t a m o re rapid r a te in the v e n tra l end of. th e rib t h a n in th e
limb bones. This is also indicated in th e n e w b o rn calf w h ich is up on
its feet w ithin a few ho u rs a f te r birth, w hich indicates t h a t a t b irth
th e leg bones are relatively m o re m a tu r e t h a n t h e ribs. T h e localiza
tion of the ricketic changes a t th e end of th e rib is a f u n d a m e n ta l p oin t
of som e im portance. In the first place, it is add itio nal evidence t h a t
rickets occurred only w h e re th e re w as g ro w th , and secondly, it has a
practical application in m a k in g ash d e te rm in a tio n s on ric k e tic bones.
Obviously, even th o u g h th e re m ay be a la rg e excess of o steo id tissue
at th e costo cho nd ral jun ctio n it will n o t be tru ly reflected in th e ash
analysis of the. entire bone since the ricketic cha n g es a re confined to
a v ery small p a r t of an o th erw ise well ossified rib.
F r a g m e n t a r y evidence indicates t h a t th e epiphyseal type o£ h ealin g
rickets occurred in one calf in this stu dy (Fig. 26).
W ith th e exception of th e blood p lasm a and possibly th e lym ph, th e
changes which occurred in th e so ft tissues of th e bo d y w e r e n o n
PATHOLOGY OF RICKETS IN DAIRY CALVES
19
specific and w ere probably the result of general im pairm ent of bc>dy
health. In one case of advanced rickets, however, muscle atroph y and
erosion of the artic u la r surfaces w ere observed in addition to the usual
sym ptom s of rickets in calves. P ittin g and erosion of the joints was
seen in several of the ricketic calves in the present investigation but
never to such a m ark e d degree as in the one calf described above
(Figs. 33 and 34). H u ty ra and M arek (9) included ulcerous d estru c
tion of the joint cartilages along w ith inflammation of the joints in
their list o f general anatomical changes of rickets in cattle. They also
noted t h a t severe rickets was accompanied by increasing emaciation
and t h a t prognosis depended upon nervous disturbances and complica
tions.
Gullickson and co-w orkers (18)^ did not m ake histological studies of
the rickets-like disease which they encountered in calves. Their g en
eral observations, however, re g ard ing gain in body weight, composition
of the blood plasm a and skeletal alterations, including erosion of the
a rticu latin g surfaces, are fundam entally in agreem ent w ith the find
ings of our w o rk on rickets in dairy calves.
T he tw o types of low vitam in D rickets observed in this study ap
pear to be histologically identical. P erhaps one should n o t expect to
find histological differences in bones from cases of low blood calcium
rickets and cases in which the concentrations of both calcium and
inorganic phosphorus are low.
A com parison of the results which were obtained in this investiga
tion w ith the reports in the literatu re indicates th at the alterations in
ricketic bone are fundam entally the same in the calf as those described
for the y o u ng r a t and for the infant. This similarity in response to a
deficiency of the antiricketic factor dem onstrates the suitability of
the dairy calf for problems of fundamental research in the n utrition
of vitam in D.
SUMMARY
This investigation was designed to determ ine the histological a ltera
tions th a t occur in the tissues of dairy calves suffering from a deficiency
of vitam in D. The study was based on 16 grade Holstein calves which
w e re selected from a group of more than 100 animals available for this
work. T he ages of the calves varied from 151 to 520 clays at the time
of death. Five of these calves w ere normal, while the rem aining 11
calves represented several stages in the severity of rickets. The d ura
tion of th e disease varied from 38 to 212 days. One case of healing
rickets was included in the study and also one case of rickets in which
muscle atro p h y and erosion of th e joint cartilage had occurred.
L o w vitam in D rickets in dairy calves was characterized by changes
in the bones which w ere apparently accompanied by decreased con
ce n tration s of calcium a n d /o r inorganic phosphorus in the blood plasma.
The costochondral junction at th e ventral end of the rib was found
to be th e best index to ricketic changes in the skeleton. M id-frontal
sections of about four mm. in thickness were tak en from the last three
inches of the v en tral end of each rib and studied by the comparison of
ro e n tg e n o g ram s, photog rap hs of specimens stained in silver n itrate
solution, and histological sections.
20
M ICH IGA N T E C H N IC A L B U L L E T IN NO. 150
R o e n tg e n o g ra m s w e re su perior to th e specim ens stained in silver
n i t r a t e fo r this study. Sections w e re p re p a re d fo r h istological s tu d y
by decalcification in 5 p er c e n t aqueous solution of p o ta ss iu m dichrom a te followed by th e paraffin im bedding tech n iq u e an d w e r e re g r e s sively stained in hem ato x y lin and eosin solutions.
D etailed descriptions w e re m ade of th e co sto cho ndral ju n c tio n s of
b o th n o rm al and rick etic ribs. In the specim ens studied, th e histolo gical
changes w e re largely confined to a re latively sm all p o rtio n of th e bone
a t th e co sto cho ndral junction.
R e ta rd e d provisional calcification of th e ca rtila g e m a t r i x ap p e a re d
to be th e fu n d a m e n tal ch ang e in rickets. T h e m o st conspicuous c h a n g es
in th e m icroscopical study, h ow ever, w e re th e irr e g u la r re m o v a l of
ca rtila g e by the em bryonic m a rr o w an d th e a c cu m u latio n of excess
osteoid tissue. T h e b ead ing of rick etic ribs ap p e are d to be cau sed b y
th e ac cum ulation of osteoid tissu e an d w a s e x a g g e ra te d in a p p e a ra n c e
on the m edial b o rd e r by increased c u r v a tu re of t h e rib in a m edial
direction. “ C upping” of th e v entral-ep ihy seal end of th e diaphysis
w as n o t a p ro m in e n t fe a tu re in ro e n tg e n o g ra m s of th e c o s to ch o n d ral
ju nctions of rick etic calves used in this study.
G ro w th w as an im p o rta n t m odify in g fa c to r in rick ets. M o re severe
rick e ts w as associated w ith m o re rapid g ro w th . W ith in a g iv en period
of time, y o u n g e r calves developed m o re florid ric k e ts t h a n did older
calves.
L o w plasm a calcium rick e ts appe are d to be histologically identical
to low calcium-low inorganic pho sp horu s rickets.
F r a g m e n t a r y evidence s u g g e s te d t h a t th e epiphyseal ty p e of h e a l
in g rickets occurs in dairy calves. E m a c ia tio n of th e body an d erosion
of the artic u la r ca rtilages occurred as com plications in one case of
rickets.
j
H istological alte ra tio n s of rick e ts in calves, as o bserv ed in th is in
vestigation, ap pear to be fun d a m e n tally th e sam e as th o se re co rd e d
in th e lite ra tu re for the in fan t and for t h e y o u n g rat.
A b n o rm a l accum ulations of bile of an o ra n g e to y ellow color and of
viscous ch a ra c te r w e re o bserved in several cases of ric k e ts a t p o s t
m o rtem . Bile stained in g e s ta occasionally occ u rre d in th e u p p e r p a r t
of th e small intestine. E n te ritis w as n o ted in a few cases.
PATHOLOGY OF RICKETS IN DAIRY CALVES
CONCLUSIONS
In the specimens studied, the following conclusions seem to be ju s
tified :
1.
A t autopsy, low vitam in D rickets in dairy calves is characterized
principally by changes in the bones. These changes may be con
veniently studied in the ventral end of the eighth rib.
2 . R etard e d provisional calcification of cartilage m atrix appears to
be the fundam ental change in rickets. The most conspicuous his
tological changes in the disease, however, are irregular removal
of cartilage and accumulation of excess osteoid tissue.
3. Accum ulations of osteoid tissue are responsible for the beading
of ricketic ribs. This beading is typically exaggerated in gross
appearance on the medial border of the rib by increased cu rv a
tu re of the rib in a medial direction.
4. G row th is an im p ortant modifying factor in rickets. The ricketic
condition is confined laYgely to the grow ing area at the end of
the bone. M ore severe rickets occurs in more rapidly growing
dairy calves. Age, because of its relation to grow th, is also a
m odifying factor in th a t younger calves develop more florid rickets
than older calves within a given period of time when maintained
under similar conditions.
5. Histological alterations of rickets in calves are fundamentally
the same as those recorded in the literature for the infant and for
the young rat.
M ICH IG A N T E C H N IC A L B U L L E T IN NO. 150
LITERATURE CITED
1.
2.
3.
4.
5.
6.
7.
8.
9.
10.
11.
12.
13.
14.
15.
16.
17.
18.
Reed, O. E., and H uffm an , C. F.— F e e d in g of c o n c e n tra te s to dairy
cattle. H e av y feeding of c o n c en trate s w ith o u t th e p ro p e r quality
of ro u g h a g e is d etrim e n ta l to th e animal. Mich. A g r. E xp. Sta.
Q u a rt. Bui., 8 : 118 (1926).
H uffm an, C. F.— M ak e stu d y of rick ets in calves. S u nshin e c o u n t
eracts deficiencies in ration s w hich w ould cause disease. Mich.
A gr. Exp. Sta. Q u a rt. Bui., 14: 42 (1931).
H uffm an, C. F., and Duncan, C. W .—-Vitamin D studies in cattle.
I. T h e a n tira ch itic value of h ay in th e ra tio n of d airy cattle. J.
D a iry Sci., 18: 511 (1935).
H uffm an, C. F., and Duncan, C. W.— V itam in D studies in cattle.
II. T h e v itam in D sparing* action of m a g n e s iu m in th e ra tio n of
d airy cattle. J. D a iry Sci., 18: 605 (1935).
D uncan, C. W., and H uffm an, C. F.—V itam in D studies in cattle.
III. Influence of solar u ltraviolet ra d ia tio n upon th e blood c h e m
istry and m ineral m etabolism of d airy calves. J. D a ir y Sci. (in
pre ss) (1936).
Bechtel, H. E., H uffm an, C. F., Duncan, C. W., and H o p p e rt, C. A.—
V itam in D studies in cattle. IV. Coi*n silage as a source of vitam in
D for d airy cattle. J. D a iry Sci. (in p re ss) (1936).
Bechdel, S. I., L a n d sb u rg , K, G., and Hill, O. J.— R ic k e ts in calves.
Pa. A gr. Exp. Sta. Bub, 291 (1933).
Rupel, I. W., B o hstedt, G., and H a r t, E. B.—V ita m in D in the
n u tritio n of th e d airy calf. Wis. A gr. Exp. Sta. Res. Bui. 115 (1933).
H u ty ra , F., and M arek , J.— Special p ath o lo g y and th e ra p e u tic s of
th e diseases of dom estic animals. 3:213-269. 3rd ed. A le x a n d e r
E g e r, Chicago. (1926).
M arek , J., and W ellm an, O.— Die rachitis. G u s ta v F isch er, J en a.
(1931).
T heiler, A.—T h e o steod ystrop hic diseases of d o m esticated anim als.
Vet. J , 90: 143, 183 (1934).
H a rris , H. A.— Bone g ro w th in h e a lth and disease. O x fo rd Univ.
P ress, L o n don (1933).
M axwell, J. P., H u, C. H., and T urnbull, H. M.—-Foetal rickets. J.
P a th . Bact., 35: 419 (1932).
P appenheim er, A. M.— T he anatom ical cha n g es w hich acco m p any
th e h ealing of experim en tal r a t rick e ts u n d e r th e influence of cod
liver oil or its derivatives. J. Exp. Med., 36: 335 (1922).
Dodds, G. S.,. and Cam eron, H. C.— S tudies on e x p e rim e n ta l rick e ts
in rats. Am. J. Anat., 55: 135 (1934).
M axim ow , A. A., and Bloom, W .—A t e x t-b o o k of h istolo gy. W. B.
S au nders Co., P hiladelphia (1930).
Leriche, R., and Policard, A.— T h e n o rm a l and p a th o lo g ical ph y sio l
ogy of bone. C. V. M osby Co., St, Louis (1928).
Gullickson, T. W ., P alm er, L. S.-, an d Boyd, W . L .— A rick e ts-lik e
disease in y o u n g cattle. Minn. A g r. Exp. Sta. Tech. Bui., 105
(1935).
PATHOLOGY OF R ICK ETS IN DAIRY CALVES
Table 1.
C alf N o.
A ge at
death
(days)
23
Calves selected for studies on bone histology.
D uration
of rickets
(days)
C -1 5 1 ...............
194
N orm al
C -1 7 6 ..............
317
“
C -2 2 9 ...............
163
C -2 3 1 ...............
161
C -2 3 3 ...............
161
C -1 4 8 ..............
Rem arks
E xposed to sum m er sun.
R eceived 5 cc. cod liver oil and 15 gm. of M gO daily.
R eceived 4 lb. of irradiated m ilk daily.
“
R eceived 4 lb. of irradiated m ilk daily.
232
212
A te 1 lb . sun-cured h a y daily.
C -1 5 5 ..............
195
145
A te 1.5 lb. sun-cured h a y daily.
C -1 7 0 ..............
326
160
R eceived 5 cc. cod liver oil daily.
C -1 7 1 ..............
327
185
R ubbed 2 cc. 250D viosterol daily upon the skin. Low Ca—•
low P rickets. U nusually poor gains in body w eight.
R eceived 5 cc. cod liver oil daily.
Received 4 lb. of irradiated m ilk daily.
Low P — low Ca rickets.
C -1 7 3 ..............
319
239
C -1 7 5 ..............
318
38
C -1 8 8 ..............
520
140
C -2 2 4 ..............
163 '
C -2 3 2 ..............
161
61
R eceived 20 lb. skim m ilk daily.
C -2 3 7 ..............
151
61
R eceived 24 lb. w hole m ilk daily.
in body w eight.
C -2 3 8 ..............
161
71
R eceived 20 lb. w hole m ilk daily.
53
Low Ca rickets.
Low Ca— low P rickets.
Low P — low Ca rickets.
R eceived 5 cc. cod liver oil and 32 gm. MgCC>3 daily.
rickets.
R eceived 15 lb. corn silage daily.
R eceived 20 lb. w hole m ilk daily.
low Ca rickets.
Low Ca
Low Ca— low P rickets.
H ealing last 23 days from
Low Ca— low P rickets.
Low Ca rickets.
Low Ca rickets.
Poor gains
M IC H IG A N TE C H N IC A L B U L L E T IN NO. 150
24
Table 2.
Growth data and blood plasma analyses.
B lo o d p lasm a
A ge
C alf N o .
(days)
C M 510)
3 -1 7 6 (3) ................................
Inorg. P
Ca
(lb.)
(per cent
norm al)
10
20
30
40
50
60
70
80
90
100
110
120
130
140
150
160
170
180
190
19 4 (2)
85
95
105
114
122
122
126
143
163
177
193
214
231
246
253
255
253
257
276
288
85
86
87
86
84
78
74
77
82
82
83
86
87
87
84
80
76
74
76
78
10
20
30
40
50
60
70
80
90
100
110
120
130
140
150
160
170
180
190
200
210
220
230
240
250
260
270
280
290
300
310
317(2)
98
108
110
121
131
142
158
176
177
202
215
233
237
230
228
250
262
277
296
311
328
345
367
373
399
416
433
440
434
453
459
470
98
98
91
91
90
90
92
95
89
93
92
94
89
81
75
79
79
79
82
83
84
86
89
88
91
92
93
92
89
90
90
91
(0 B orn M arch 4, 1932— m ale.
(2) Slaughtered.
(3) B orn M a y 29, 1932— m ale.
Mg
(cm .)
(per cen t
norm al)
7 6 .8
101
1 2 .4
1 2 .3
6 .6 9
6 .1 0
2 .3 7
2 .3 0
8 1 .2
100
1 3 .5
1 3 .6
6 .9 1
7 .5 3
2 .0 9
2 .1 1
8 5 .2
99
i3 .6
1 4 .1
1 5 .4
8 .0 6
7 .4 0
8 .2 3
2 .3 4
2 .7 8
2 .7 5
9 2 .5
102
9 7 .8
102
1 4 .1
1 2 .8
7 .5 3
8 .3 9
3 .4 7
2 .3 3
9 9 .0
99
1 3 .0
1 2 .6
1 2 .8
1 3 .8
6 .0 1
8 .1 2
7 .9 6
7 .4 4
2 .4 5
2 .4 0
2 .5 7
2 .5 8
7 7 .7
101
1 3 .8
1 5 .0
8 .8 7
8 .8 3
2 .4 0
2 .4 7
8 1 .7
100
1 3 .9
1 3 .3
7 .6 2
6 .7 9
2 .3 6
2 .0 7
8 7 .2
101
(m g. per 100 cc.)
1 3 .9
7 .1 4
1 .9 8
•1 2 .9
1 3 .8
6 .7 2
8 .6 2
2 .3 7
1 .6 3
9 1 .3
99
96
9 6
1 0 .3
8 .3 3
5 .1 7
5 .4 8
2 .9 1
9 4 .3
9 5 .5
94
1 1 .4
1 0 .6
6 .6 9
7 .5 8
2 .7 4
2 .8 0
9 9 .8
95
ii.2
1 1 .8
1 1 .5
7 .7 1
6 .2 2
7 .3 5
2 .7 4
2 .6 2
3 .3 1
1 0 1 .2
94
1 0 .8
9 .9
6 .2 8
6 .3 8
2 .6 0
1 0 7 .8
98
1 0 .9
1 0 .7
1 0 .5
6 .8 7
6 .6 9
6 .2 2
2 .4 4
3 .6 8
2 .6 7
1 1 0 .2
99
1 1 .2
1 0 .8
6 .7 9
6 .4 1
3 .5 2
2 .1 7
PATHOLOGY OF RICK ETS IN DAIRY CALVES
Table 3.
25
Growth data and blood plasma analyses.
Blood plasm a
A ge
W eight
H eight
Calf N o,
Ca
(days)
C -2 2 9 0 ) ...............................
C-231 (-1) ...............................
C -233(4) ...............................
(*)
(-)
(■’)
O)
(5)
B orn October
Slaughtered.
B orn O ctober
B orn O ctober
Slaughtered.
(lb.)
(per cent
normal)
10
20
30
40
,50
60
70
80
90
100
110
120
130
140
150
160
163(2)
80
92
100
110
129
148
165
179
197
214
233
246
256
268
284
304
312
80
83
83
83
89
94
96
96
99
99
100
99
96
94
94
96
97
10
20
30
40
50
60
70
80
90
100
110
120
130
140
150
160
161(»)
97
106
116
126
144
163
181
192
202
210
228
239
236
237
249
264
265
97
96
96
95
99
104
106
103
101
97
98
96
89
83
82
83
83
10
20
30
40
50
60
70
80
90
100
110
120
130
140
150
160
161(6)
104
114
123
136
146
162
174
192
206
224
245
252
278
309
337
354
104
103
102
102
101
103
102
10.3
103
104
105
101
104
109
112
111
(per cent
normal)
(cm.)
7 1 .2
95
7 5 .0
96
8 1 .8
98
8 6 .7
96
8 8 .0
93
7 4 .5
'
Mg
Inorg. P
(mg. per 100 cc.)
1 2 .9
1 3 .8
1 3 .4
1 2 .9
1 2 .2
1 2 .3
1 2 ,0
1 2 ,3
1 3 .2
12.1
1 1 .8
12.1
8 .7
12.1
1 1 .7
1 1 .8
8 .7 4
8 .5 6
7 .9 1
8 .3 9
8 .1 2
7 .5 8
7 .0 9
6 .4 1
7 .7 7
7 .2 7
8 .0 1
9 .2 9
7 .5 8
9 .2 4
9 ,6 9
9 .1 9
2 .2 5
2 .2 1
3 .2 3
2 .5 5
2 .4 9
2 .7 2
2 .5 9
1 .9 0
2 .2 3
2 .3 2
2 .2 6
2 .4 1
2 .6 2
2 .5 5
2 .2 5
2 .9 5
101
1 2 .6
6 .1 9
2 .3 3
7 8 .2
102
8 4 .8
103
8 8 .8
100
9 4 .2
101
9 5 .6
98
1 1 .7
1 2 .6
1 2 .3
1 1 .5
1 1 .6
9 .7
12 .1
1 1 .8
1 0 .7
12 .3
1 1 .4
1 2 .7
1 1 .5
1 1 .7
10.9
6 .9 8
8 01
8 .2 2
7 .8 6
8 .2 5
8 .3 3
7 .8 6
7 .7 4
7 .0 2
8 .3 9
8 .7 4
7 .9 4
8 .6 8
9 .9 2
8 .0 6
2 .8 3
3 .1 7
2 .5 2
3 .1 7
2 .7 7
2 .4 9
2 .0 7
2 .4 5
2 .3 8
2 .9 0
2 .4 3
2 .5 8
2 .3 2
2 .9 1
3 .1 8
1 3 .4
1 1 .8
7 .5 3
8 .3 3
2 .6 9
2 .7 5
1 3 .4
12 .7
1 2 .7
1 1 .9
1 1 .7
12.1
1 2 .0
12 .3
1 2 .7
1 2 .9
1 2 .4
11.6
1 1 .8
8 .2 8
8 .5 6
7 .7 8
8.01
7 .4 0
7 .7 7
7 .2 3
7 .0 3
8 .5 0
6.31
8.01
8 .2 3
8 .5 3
2 .2 5
2 .6 1
2 .6 3
2 .6 7
2 .4 1
2 ,2 7
2 .4 5
2 .4 8
2 .9 1
2 .6 3
2 .7 5
2 .1 7
2 .3 2
7 3 .5
100
7 7 .7
101
8 1 .0
99
8 6 .2
97
9 2 .8
99
9 7 .3
99
16, 1933— m ale.
25, 1933— m ale,
28, 1933— male.
P ittin g of articulating surfaces of femur and humerus.
•
M IC H IG A N T E C H N IC A L B U L L E T IN NO. 150
Table 4.
Growth data and blood plasma analyses.
B lo o d p la sm a
A ge
w e ig in
C alf N o.
(days)
-1 4 8 0 ).
155 (r>) .
10
20
30
40
50
60
70
80
90
100
110
120
130
140
1.50
1 6 0 (2)
170
180
190
200 0 )
210
220 0 )
230
232(5)
10
20
30
40
50
60
70
80
90
100
11 0
120
1 3 0 (0
140
150
160
170
180
190
1 9 5 (8)
(0
(0
C)
(0
B orn M arch l ,
A ge 156 days.
A ge 197 days.
A ge 216 d ays.
(lb.)
(per cent
norm al)
91
98
110
118
129
144
154
166
171
171
169
168
165
165
165
167
172
170
171
175
180
190
187
91
89
91
89
89
92
90
89
86
79
73
67
62
58
55
53
52
49
47
47
46
47
45
88
102
115
123
134
143
149
157
166
171
177
200
204
199
203
206
213
217
221
223
88
92
95
92
92
91
87
85
83
79
76
80
77
70
67
65
64
62
61
60
(cm .)
(per cent
norm al)
7 7 .5
101
8 3 .5
102
8 7 .0
Mg
Inorg. P
Ca
(m g. per 100 cc.)
2 .4 0
2 .2 3
1 2 .5
1 1 .9
1 1 .3
1 1 .4
1 0 .1
5 .6 1
4 .4 6
5 .8 4
6 .2 2
5 .4 3
2 .6 3
2 .1 5
100
1 0 .0
1 1 .7
1 1 .8
5 .0 4
5 .3 2
4 .0 9
2 .2 8
2 .6 7
3 .3 1
8 7 .7
96
1 2 .2
1 1 .8
3 .9 5
2 .7 6
3 .0 2
2 .8 5
8 5 .8
89
3 .1 2
2 .5 8
3 .9 3
2 .6 6
1 .7 3
2 .8 0
8 4 .5
84
1 2 .0
1 0 .4
9 .7
9 .9
9 .6
2 .9 6
3 .2 3
2 .2 3
2 .1 5
1 2 .3
8 .5
7 3
5 .0 0
4 .0 5
4 .1 3
2 .1 2
1 .2 2
0 .9 9
6 .8 3
8 .2 3
7 .2 7
2 .2 9
2 .6 9
2 .8 0
D ow n
7 7 .5
103
1 3 .7
1 3 .5
1 2 .7
8 3 .7
105
1 0 .9
6 .2 8
2 .3 1
8 7 .8
103
9 .3
9 .9
9 .3
7 53
7 .6 2
8 .3 3
2 13
2 .2 4
2 .6 9
9 0 .0
100
9 .8
9 .7
7 .5 3
6 .5 4
2 .0 5
2 .4 7
9 1 .8
97
9 .3
8 7
7 .3 1
6 .3 2
2 .1 9
9 3 .7
94
8 .1
8 .4
8 .9
7 9
5 .8 1
5 .2 7
8 .5 0
7 .9 1
2 .0 7
1 .7 7
1 .6 7
1 .6 6
1932— m ale.
Legs bow ed. J o in ts sw ollen .
U nable to w alk to scales.
U nable to sta n d lo n g enough to drink m ilk.
( 5 ) C onvulsion . Slau ghtered .
(«) B orn M arch 10, 1932— m ale.
(») A ge 134 d ays. L igh t c o n v u lsio n .
(8) Slau ghtered .
PATHOLOGY OF RICKETS IN DAIRY CALVES
T a b l e S.
27
G r o w t h d a t a a n d b lo o d p l a s m a a n a ly s e s .
B lood plasm a
A ge
Calf N o.
C - 1 7 0 0 ...............................
C -188(6) ...............................
(i)
0
0
(i)
(*)
(»)
Ca
(per cent
normal)
(days)
(lb.)
10
20
30
40
50
60
70
80
90
100
110
120
130
140
150
160
170
180
190
200
210
220
230
240
250
260
270
280
290
3000
310
3200
3260
97
96
103
111
118
128
140
146
153
161
178
189
194
198
208
220
230
243
269
286
301
326
345
355
377
396
411
429
448
462
463
461
473
97
87
85
83
81
82
82
79
77
74
77
76
73
70
68
69
69
70
74
76
77
81
84
84
86
88
88
90
92
92
91
89
89
360
370(6)
380
3900
400
410
420 (s)
430
440(6)
450
460
470
480
490(16)
500
510
520(H)
500
525
524
523
549
520
538
540
532
520
529
521
509
496
466
436
406
90
93
92
91
94
88
90
90
88
85
85
82
79
76
71
66
61
Born M a y 1, 1932— m ale.
G ettin g stiff in legs. B ack arched.
V ery stiff in legs.
Slaughtered.
B o m N ovem ber 14, 1932— male.
Joints enlarged. Rear legs stiff.
(7)
(s)
(9)
(1(l)
(»)
(cm.)
(per cent
normal)
7 6 .2
100
Inorg. P
Mg
(mg. per 100 cc.)
1 1 .6
6 .6 9
• 2 .1 2
12 6
1 3 .6
5 .8 1
7 02
5 .7 4
1.99
2 .1 4
2 .6 2
13 5
14.1
7 .7 6
6 .6 5
2 74
2 88
1 3 .2
1 1 .9
6 .3 8
7 35
2 45
2 .5 1
i2 .2
1 0 .6
5 .9 5
4 90
5 .1 7
2 .6 9
1 .9 0
2 .6 3
9 6
8 .7
5 .1 0
5 .5 8
2 .0 8
2 .1 3
8 0 .2
98
8 4 .0
97
8 8 .2
96
9 0 .7
94
9 2 .7
91
9 8 .2
94
8 7
10 5
9 .5
6 .5 4
6 69
7 .5 3
2 .5 7
2 .9 3
2 .7 5
1 0 2 .7
95
8 8
9 .3
6 .1 3
5 .3 2
2 .6 0
2 .6 9
1 0 3 .7
95
8 1
7 .3
8 3
6 41
5 .7 1
5 .1 2
2 .6 2
2 .5 8
2 .3 3
1 0 7 .7
96
8 .0
9 .4
5 .3 4
4 .7 3
2 .7 8
2 .5 9
1 0 .8
9 .5
8 .3
1 0 .3
8 .6
8 .2
8 .3
7 .4
1 0.3
7 .8
8 .2
7 .9
8 .2
8 .5
8 .6
8 .0
8 .1
8 .1 2
8 .6 2
9 .1 9
5 .9 3
6 .6 2
8 .2 8
6 95
7 .2 5
5 08
4 .6 3
5 17
5 63
4 .3 7
4 19
3 29
2 96
3 .5 0
3 .1 0
3 .5 5
2 .5 6
2 .2 6
2 .90
2 .6 1
2 .5 8
2 .6 0
2 .25
2 .8 3
2 . 67
2 .45
2 .8 3
2 59
2 49
2 49
2 .3 7
1 0 9 .8
96
1 1 2 .8
98
1 1 4 .8
98
1 1 4 .2
96
1 1 3 .8
95
K nees bowed.
Stands cross legged in rear. D rags rear feet.
Very stiff. Lies down m ost of time.
A ge 487 days. In coma,
D ied.
28
M ICH IG A N T E C H N IC A L B U L L E T IN NO. 150
Table 6.
Growth data and blood plasma analyses.
B lo o d p la sm a
A ge
n e ig u i
W eig h t
(days)
C -1 7 1 0 )1................................
(lb.)
(per cent
norm al)
(cm .)
(per cent
norm al)
143
151
84
79
80
83
85
oO
83
SI
8 3 .8
97
158
161
168
73
69
67
8 5 .2
95
140
181
179
64
59
8 7 .8
92
170
180
190
280
290
300
310 (3)
320
327 (4)
181
179
186
198
210
215
219
225
230
241
257
264
269
280
284
283
283
54
51
51
53
54
54
53
53
52
53
55
55
55
56
56
54
54
10
20
30
40
50
60
70
80
90
100
110
120
130
140
150
160
163 (0)
93
106
117
127
143
162
177
192
213
229
244
264
283
298
312
329
342
93
96
97
95
99
103
103
103
107
106
105
106
106
105
103
104
107
10
20
30
40
50
60
70
80
90
100
110
120
210
220
240
250
260
84
87
97
110
123
7 8 .0
103
99
Mg
Inorg. P
Ca
C alf N o.
(m g. per 100 cc.)
1 2 .0
6 .1 6
2 .2 4
1 2 .9
1 2 .2
1 1 .9
5 .9 3
6 .9 5
6 .9 1
1 .9 1
1 .9 4
2 .4 0
1 2 .5
1 3 .4
6 .2 5
6 .8 3
2 .6 3
3 .1 0
i 3 .1
1 2 .5
1 2 .5
6 .2 2
7 .7 6
6 .2 5
2 .3 8
2 .3 3
2 .5 8
1 1 .2
5 .0 0
4 .8 4
2 .1 3
2 .2 9
4 .2 2
3 .5 0
2 .8 1
1 .9 6
1 .6 1
2 ,2 9
8 8 .3
87
9 .0
9 .0
8 .0
8 9 .2
85
8 .6
9 .1
4 .4 7
7 .0 2
2 .3 6
2 .7 2
9 .9
9 0 .5
84
8.0
7 .5
4 .6 3
6 .2 5
5 .6 8
2 .5 3
2 .5 1
2 .6 0
9 1 .7
84
6 .4
7 .3
6 .4 8
4 .5 8
2 .8 2
2 .1 7
9 5 .0
85
8 .6
8 .6
4 .0 8
3 .8 1
2 .8 0
3 .0 2
7 6 .8
102
7 9 .0
98
83 5
98
9 4 .5
105
9 6 .5
102
1 2 .5
1 3 .0
1 2 .5
1 2 .1
1 2 .0
1 2 .1
1 1 .4
1 1 .1
1 0 .0
7 .6
8 .0
8 .5
1 1 .1
7 .4
9 .8
9 .6
8 .4 5
7 .9 1
6 .9 8
6 .9 8
6 .4 1
6 .3 2
6 .1 0
7 .0 0
7 .0 6
7 .7 9
6 .7 6
7 .1 8
1 0 .2 4
8 .2 3
7 .9 5
7 .0 2
2 .2 7
2 .7 5
2 .1 5
2 .6 5
2 .3 9
2 .9 6
2 .6 2
2 .3 7
2 .5 6
2 .1 6
2 .1 5
2 .2 4
2 .3 7
2 .5 1
2 .3 4
2 .9 7
(!) B orn M a y 3, 1932— fem ale.
( 2 ) A ge 275 d a y s. L igh t con v u lsio n s.
(3) A ge 315 d a y s. Legs stiff. D ifficu lt to rise to feet.
(4) B lo a t p robab le cause of d ea th .
( 6) B orn O ctober 14, 1933— m ale.
( 3) S lau ghtered .
PATHOLOGY OF RICK ETS IN DAIRY CALVES
Table 7.
29
Growth data and blood plasma analyses.
Blood plasm a
Age
W eight
H eight
Calf N o.
-7
W
o
(days)
C -232(" )...............................
(1)
(2 )
(*)
(<)
Inorg. P
Ca
(lb.)
(per cent
normal)
10
20
30
40
50
60
70
80
90
100
110
120
130
140
150
160
170
180
190
200
210
2 2 0 (2)
230
240
250
2 6 0 (3)
270
280
290
300 C)
310
319«
92
102
108
109
115
117
117
123
122
129
140
153
161
176
187
200
221
239
250
262
265
290
316
324
336
342
346
355
347
351
342
323
92
92
89
82
79
75
68
66
61
60
60
61
60
62
62
63
66
68
69
70
68
76
77
76
77
76
74
74
71
70
67
62
10
20
30
40
50
60
70
80
90
100
110
120
130
1400
150
160
161 (s)
100
109
120
134
148
158
162
174
183
201
229
231
245
259
276
275
100
98
99
101
102
101
95
94
92
93
98
93
92
91
91
87
B orn M a y 8, 1932— m ale.
Stiff in front legs,
K nees bow ed. Legs v ery stiff.
Stiff in legs. B arely able to rise to feet.
Mg
(cm.)
(per cent
normal)
7 4 .8
100
1 4 .3
1 4 .3
6 .8 7
6 .6 9
2 .3 2
2 .1 9
7 8 .0
97
1 3 .9
1 3 .8
1 4 .8
5 .1 0
4 .5 5
7 .8 1
3 .4 3
2 .0 9
2 .5 3
8 1 .0
95
12.8
12.5
4 .4 6
4 .3 6
3 .0 2
1 .6 4
8 1 .8
91
1 2 .0
12.9
1 2 .8
8 4 .5
89
5 .8 7
5 .5 6
4 .4 3
5 .9 0
1 .5 4
2 .3 4
2 .0 5
2 .8 6
8 9 .3
89
9 3 .3
90
9 8 .5
92
10 1.5
93
1 0 4 .2
94
7 6 .3
104
7 9 .0
103
8 5 ,7
105
9 1 .3
103
9 4 .0
101
9 6 .3
98
(mg. per 100 cc.)
10.3
6 .0 4
1.76
9 .9
9 .3
10.2
6 .3 2
5 .8 4
5 .5 3
1 .7 8
2 .2 8
1 .8 0
11 .3
1 1 .2
6 .0 7
5 .6 3
2 .7 2
2 .7 4
1 0 .8
1 0 .4
10.1
6.01
4 60
5 .8 1
2 .8 8
2 .1 0
2 .4 5
9 .7
1 0 .9
9 .0
9 .0
5 33
4 .2 8
5 .1 7
3 .9 0
2 33
3 .4 7
2 .4 3
2 .9 3
1 3 .3
1 2 .8
13.1
12.9
1 1 .8
1 2 .0
9 .9
1 0 .0
9 .4
7 .7
7 .7
8 .0
8 .5
8 .0
8 .0
7 .9
6 .0
8 .0 1
8 .7 4
8 .5 6
8 .3 9
8 .8 7
7 .9 4
7 .2 3
7 .4 9
7 .1 9
6 .9 1
6 .1 8
6 .6 5
5 .9 9
5 .6 3
5 .7 9
4 .9 6
4 .9 8
2 .5 8
2 .9 0
2 .6 0
2 .4 6
2 .6 3
3 .2 5
2 .1 6
2 .0 7
1 .8 8
2 .9 7
2 .7 1
3 .1 6
2 .7 7
2 .5 7
2 .2 5
2 .5 4
2 .5 3
(6) Slaughtered.
(') Born October 28, 1933— m ale.
0 ) L egs bowed.
(8) Slaughtered. Slight p ittin g of articular surface
on ball of femur.
30
M IC H IG A N T E C H N IC A L B U L L E T IN NO. 150
Table 8.
Growth data and blood plasma' analyses.
B lood p lasm a
A ge
C alf N o.
(lb.)
(per cent
norm al)
10
20
30
40
50
60
70
80
90
100
110
120
130
140
150
160
170
180
190
200
210
220
230
240
250
260
270
280
290
300
310
3 1 8 (2)
100
110
114
132
147
159
176
193
209
240
244
266
271
292
310
329
342
352
380
409
422
439
461
468
479
505
516
535
537
553
555
565
100
99
94
99
101
101
103
104
105
111
105
107
102
103
103
104
103
101
105
109
108
109
112
110
109
112
111
112
110
108
109
109
10
20
30
40
50
60
70
80 0
90
100
110
120
130
140(6)
1 5 0 (6)
1510
102
122
134
142
155
165
181
186
190
197
202
214
235
244
238
102
no
111
107
107
105
106
100
95
91
87
86
88
86
79
(days)
C -237 (3) ................................
(0
(2)
(3)
(<)
Inorg. P
Ca
B orn M a y 28, 1932— m ale.
Slaughtered.
B orn N ovem b er 10, 1933— m ale.
C on v u lsio n at 82 d a y s of age and very irritable
a t 83 days of age.
(cm.)
(per cent
norm al)
7 7 .5
101
8 4 .3
103
8 8 .8
102
9 3 .2
101
Mg
(m g. per 100 cc.)
1 4 .0
7 .4 9
2 .7 4
1 4 .0
1 5 .0
7 .2 3
7 .4 9
2 .3 8
3 .7 1
1 4 .4
1 3 .3
1 2 .6
1 2 .7
1 3 .0
6 .7 9
8 .0 1
7 .5 8
5 .9 5
7 .6 2
1 .9 4
2 .1 6
2 .3 3
2 .3 6
2 .2 2
1 1 .4
6 .9 1
7 .6 2
2 .7 2
1 .9 8
1 1 .3
1 1 .5
1 2 .2
7 .2 7
7 .6 2
7 .3 5
1 .6 1
2 .6 9
2 .5 9
1 2 .1
1 2 .7
8 .0 6
7 .0 6
3 .1 0
3 .0 2
9 7 .5
100
1 0 2 .2
100
1 0 5 .5
100
1 0 9 .5
102
1 1 .5
9 .8
1 1 .0
7 .8 1
8 .7 4
7 .8 1
2 .8 8
2 .7 5
2 .6 0
1 1 4 .5
104
10 2
9 .6
8 .7 4
8 .2 8
2 .5 5
3 .3 1
9 .5
8 8
9 .4
9 .1 3
9 .7 7
8 .5 0
2 .9 1
3 .2 4
3 .3 1
1 2 .2
12 4
1 3 .0
1 3 .5
1 1 .5
1 1 .7
1 0 .1
6 .4
8 5
7 2
8 .8
8 8
6 .7
6 .5
8 .5 6
6 .3 8
6 .8 3
6 .5 8
6 .6 9
6 .2 1
7 .6 2
7 .6 6
6 76
7 .6 7
8 .0 6
8 68
8 06
7 .4 5
2 .2 5
2 80
2 .7 2
2 .3 3
2 .2 0
2 .0 7
1 .7 0
1 .9 9
1 .7 5
1 .6 5
2 .4 5
2 .2 7
1 .83
1 .8 4
7 6 .7
102
8 1 .3
101
8 9 .8
103
9 0 .3
98
9 4 .7
98
(5) C onvulsion s at 139, 140 and 143 d a y s of age.
CO C onvulsion . U n ab le to sta n d on rear legs.
(7) Slaughtered. F ractu red verteb ra.
PATH O LOG Y OF RICK ETS IN DAIRY CALVES
T a b l e 9.
31
G r o w t h d a t a a n d b lo o d p la s m a a n a ly s e s .
Blood plasm a
A ge
W eight
H eight
Calf N o.
Ca
(days)
C-238C1) ...............................
(lb.)
10
20
30
40
50
60
70
80
90
100
110
120
130
140
150
160
161 P)
(>) B o m N ovem ber 10, 1933— male.
(2) Slaughtered.
113
109
122
130
145
160
184
205
229
253
269
290
314
336
353
370
(per cent
normal)
113
98
101
98
100
102
107
110
115
117
116
116
118
118
117
116
(cm.)
(per cent
normal)
78 0
104
8 4 .0
105
9 3 .2
107
9 6 .7
105
1 0 3.3
107
Mg
Inorg. P
(mg. per 100 cc.)
1 3 .3
12.6
1 2 .5
1 2 .0
1 2 .7
11 .6
9 .3
9 ,5
8 .5
8 .7
8 .6
8 .2
8 .1
8 .0
8 .4 7
7 .6 2
6 .9 2
6 .1 3
6 .8 9
7 .4 9
6 .9 0
7 .0 2
6 .7 9
7 .1 6
6 .3 5
5 .6 4
6 .2 2
5 .9 2
2 .6 1
2 .4 7
2 .4 5
2 .3 7
2 .2 2
3 .1 1
2 .1 7
2 .1 2
2 .2 0
2 .1 6
2 .2 4
2 .2 3
2 .4 7
2 .4 7
32
MICHIGAN T E C H N IC A L B U L L E T IN NO. 150
F i g . 1. C a lf C -148. S e v e r e r ic k e t s . S h o w s e m a c ia t io n , h u m p i n g o f b a c k , s w e l li n g o f j o in t s ,
k n u c k li n g o f p a s t e r n s , a n d b o w in g o f l e g s . T h is c a lf w a s u n a b le to w a l k a n d w a s o n l y a b le
t o s t a n d lo n g e n o u g h to b e p h o t o g r a p h e d .
F i g . 2. C a lf C -238. T h is c a lf r e c e iv e d a b a s a l r ic k e t o g e n ic
w h o le m ilk .
S h o w s a b s e n c e o f d e f in i t e c lin ic a l s y m p t o m s
s e v e r e r ic k e t ic c h a n g e s in t h e r ib a s s h o w n in F i g s . 12,
a n a l y s e s in T a b le 9. T h is c a s e i s a g o o d ill u s t r a t io n o f
w h ic h m a y b e p r o d u c e d m t h e r ib b e f o r e t h e l e g b o n e s s h o w
r a tio n s u p p l e m e n t e d w i t h n a t u r a l
o f r ic k e t s .
N o te , h o w ev er
th e
20, a n d 26. N o t e ’a l s o t h e b lo o d
t h e e x t e n s i v e r ic k e t ic a l t e r a t i o n s
c li n i c a l e v id e n c e o f t h e d i s e a s e .
PATHOLOGY
OP
PICKETS
IX
DAIPV
( ALYF.S
F i g . 3. C a lf C -231. N o r m a l. S h o w s : r e s t in g c a r t ila g e , A ; c e ll p r o life r a tio n , B ; c e ll
g r o w t h , C ; m a tu r e c a r t i la g e c e lls w it h c a lc ifie d m a tr ix b e t w e e n t h e r o w s of c e lls , D ;
v a s c u la r b u n d le , E .
( T h is a n d a ll s u c c e e d in g p h o t o m ic r o g r a p h s illu s t r a t e m id - f r o n ta l
s e c t io n s t h r o u g h t h e le f t e ig h t h c o s to c h o n d r a l ju n c tio n . T h e d ia p h y s is is to t h e r e a d e r ’s
r ig h t, e x c e p t in F i g . 14 w h e r e i t is t o th e b o t to m o f t h e p a g e ) . 120X.
F ig
4 A r e a A in F i g . 3. S h o w s : m a tu r e c a r tila g e c e ll, A ; is o g e n o u s
p a ir of im m a tu r e c a r t ila g e c e lls , B; c a r tila g e m a tr ix , C. 460X.
34
M ICHIGAN T E C H N IC A L B U L L E T IN NO. 150
F i g . 5. C a lf C -233. N o r m a l. S h o w s : o s t e o c l a s t s , A ; c o r e o f c a lc i f i e d c a r t i l a g e
m a t r i x , B ; d e p o s itio n o f g r a n u l e s o f in o r g a n ic s a l t s in s t r i a t i o n s , C . 4 6 0 X .
F i g . 6. F r o m n o r m a l s p e c im e n . S h o w s : o s t e o c l a s t s , A ; p e r io s t e u m , B . N o t e l a r g e n u m b e r s
o f o s t e o c l a s t s h e r e in t h e r e g io n w h e r e t h e t r a b e c u l a e m e e t t h e p e r io s t e u m . 1 2 0 X .
PATHOLOGY OF RICKETS IN DAIRY CALVES
F i g . 7. C a lf C -231 . N o r m a l a d v a n c e m e n t of e m b r y o n ic m a r r o w . S h o w s : m a tu r e
c a r t ila g e c e lls , A ; c a lc ifie d m a tr ix b e t w e e n m a tu r e a n d d e g e n e r a tin g c a r t ila g e c e lls , B ;
c o a g u la t e d b lo o d e le m e n t s in s m a ll s a c -lik e a r e a s in z o n e of c a r t ila g e r e m o v a l, C;
t o n g u e o f e m b r y o n ic m a r r o w , D ; t r a b e c u la e of c a lc ifie d t is s u e , E ; o s t e o c l a s t s , F ;
o s t e o b l a s t s , G . 120X .
F ig 8
C a lf C -231. N o r m a l. S h o w s : v a s c u la r b u n d le s in r e s t in g c a r tila g e , A ; a r e a
o f p r o lif e r a t io n a n d c e ll r o w s , B ; c a lc ific a tio n a t j u n c tio n o f c a r tila g e w ith th e d ia p h y s is ,
C ; t r a b e c u la e o f c a lc ifie d s u b s t a n c e D ; m a r r o w , E . N o t e th e r e g u la r a n d e v e n m a n n e r
in w h ic h t h e d ia p h y s is jo in s w it h th e c a r tila g e n o u s e n d of th e b o n e . 30A.
36
M ICHIGAN T E C H N IC A L B U L L E T IN NO. 150
F i g . 9. C a lf C -175. M ild r ic k e t s . S h o w s : r e ta r d e d a n d ir r e g u l a r p r o v i s i o n a l c a lc i f i c a t i o n o f
c a r t i la g e m a t r i x , A ; h e a v i e r d e p o s it s o f g r a n u l e s o f in o r g a n ic s a l t s in p e r ip h e r y o f l o n g it u d i n a l
b a r of c a r t i la g e m a t r i x , B ; c o a g u l a t e d b lo o d e l e m e n t s in s a c - l ik e s t r u c t u r e , C ; o s t e o c l a s t , D ;
o s t e o b l a s t , E ; e m b r y o n ic m a r r o w , F . 46 0 X .
C alf C -170. M ild r ic k e t s . S h o w s : e x t e n s i o n o f t o n g u e s o f c a r '
v e r t e b r a l ly in to t h e d i a p h y s is , A . 3 0 X .
PATHOLOGY OF RICKETS IN DAIRY CALVES
F i g . 12.
F i g . 11. C a lf C -233. N o r m a l. S h o w s : r e s t i n g c a r tila g e , A ; a r e a o f p r o life r a tio n and c a r t i
la g e c e ll r o w s , B ; ju n c tio n o f t h e d ia p h y s is w it h th e c a r tila g e n o u s en d o f th e r ib — th e s li g h t
s e p a r a tio n n o t e d h e r e is an a r tif a c t , C ; b a r s o f b o n e t r a b e c u la e — n o te t h e ir d a r k s ta in , h o w
th e y e x t e n d in a d ir e c tio n p a r a lle l to th e lo n g a x is o f th e b o n e , a n d h o w t h e y d e c r e a s e in
n u m b e r a n d b e c o m e h e a v ie r f u r th e r in th e d ia p h y s is , D ; m a r r o w , E ; t h ic k e n in g o f p e r i
c h o n d r iu m , F . 5X .
F i g . 12. C a lf C-238. S e v e r e r ic k e t s . S h o w s : r e s t i n g c a r t ila g e , A ; a rea o f p r o life r a tio n a n d
c e ll r o w s , B ; in c r e a s e d d e p th o f z o n e o f m a tu r e c a r t ila g e c e lls , B 1; a r e a of c a r tila g e r e m o v a l—
n o t e i r r e g u la r it y in t h is a r e a , C ; s h o r t, s tu b b y t r a b e c u la e , D ; s a c - lik e s t r u c t u r e s iso la te d in
ir r e g u la r ly - t h in n e d c a r t i la g e , E : e m b r y o n ic m a r r o w a n d o s te o id t is s u e , F . N o t e ir r e g u la r
a d v a n c e m e n t of th e m a r r o w a n d its e x te n s io n v e u t r a lly in th e a x ia l r e g io n o f t h e b o n e . SX.
37
33
M ICH IGA N T E C H N IC A L B U L L E T IN NO. 150
c*
* .
F i g . 13. C a lf C -233. N o r m a l.
S h ow s:
o s t e o c l a s t , A ; o s t e o b l a s t b o r d e r in g a t r a b e c u l a o f
b o n e — n o t e e c c e n t r ic a l ly p la c e d n u c le i, B ; o s t e o b l a s t s u r r o u n d e d b y o s t e o i d t i s s u e a n d a l s o p a r t ly
s u r r o u n d e d b y d a r k g r a n u l e s o f in o r g a n ic s a l t s — s u c h c e ll s a r e g r a d u a l ly c h a n g i n g i n t o o s t e o c y t e s , C ; o s t e o i d t i s s u e , D : c o r e o f c a lc i f i e d c a r t i l a g e m a t r i x a s y e t u n r e m o v e d b y c h o n d r o l y s is ,
E . 460X .
l ' i g . 14.
C a lf C -151. N o r m a l. S h o w s : g r o u p s o f o s t e o b l a s t s a s t h e y o c c u r o n t h e b o r d e r s
o f t h e la r g e r t r a b e c u la e m t h e v e n t r a l e n d o f t h e d i a p h y s is , A . 46QX.
PATHOLOGY OF RICKETS IN DAIRY CALVES
39
F ig 1. 15. C a lf C -233. N o r m a l. S h o w s : e m b r y o n ic m a r r o w , A ; t r a b e c u la e o f b o n e , B ; o s t e o
b la s t s b o r d e r in g tr a b e c u la e , C ; o s t e o c l a s t s o n e n d s o f t r a b e c u la e w h e r e t h e y a r e a s s o c ia t e d
w it h t e r m in a l e r o s io n o f th e c a lc ifie d t r a b e c u la e , D . 120X.
F i g . 16.
A r e a D in F i g . 15.
S h ow s:
te r m in a l e r o sio n b y o s t e o c la s t s , D .
575X .
MTCTTTCAN T E C H N IC A L B U L L E T IN NO. ISO
40
.
t
j .*»
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.A ,
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4
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1
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F i g . 17.
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m 1 Cm .
' 1.
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1 *
l> .
C a lf C -15S, S e v e r e r ic k e t s . S h o w s : r e s t i n g c a r t i la g e , A ; c e ll p r o lif e r a t io n , B ;
c e ll g r o w t h , C ; e m b r y o n ic m a r r o w , D ; w a l l o f b lo o d v e s s e l , E . 1 2 0 X .
m
( j
F i g 18. A r e a A in F i g . 17. S h o w s : i n c r e a s e d n u m b e r o f i s o g e n o u s p a ir s o f c a r t i l a g e c e l l s in
s h r u n k e .^ o n d .t i o n , B . N o te d e c r e a se m a m o u n t of m a tr ix a n d c o m p a r a tiv e a b se n c e o f m a tu r e
PATHOLOGY OF RICKFTS IX DAIRY CALVES
41
P i g . 19. C a lf C -237. A d v a n c e d r i c k e t s .
S h o w s:
l a r g e n u m b e r of s a c - l ik e s t r u c t u r e s , A ;
l e n g t h e n e d r o w s of m a t u r e c a r t i l a g e c e lls , B ; i r r e g u l a r l y s h a p e d t r a b e c u l a e of c a lc if ie d t i s s u e , C ;
a r t i f a c t , D . 30X .
F i e 20
C a lf C -238
S e v e r e r i c k e t s . S h o w s : s a c - l ik e s t r u c t u r e s , A ; e n o r m o u s ly in c r e a s e d
d e p t h o f z o n e o f m a t u r e c a r t i l a g e c e lls , B ; i r r e g u l a r l y s h a p e d t r a b e c u l a e of b o n e C ; e m b r y o n ic
m a r r o w a n d o s te o i d t i s s u e D . N o t e i r r e g u a r a d v a n c e m e n t , v e n t r a l l y , o f t h e e m b r y o n i c m a r r o w
F t h e a x F l r e g i o n o f t h e b o n e . C o m p a r e w i t h F i g . 19, C a lf C -237, w h ic h h a d r i c k e t s o f s im il a r
s e v e r i t y . ' a s i n d i c a t e d b y t h e b lo o d p i c t u r e f o r a b o u t t h e s a m e l e n g t h o f t im e b u t m a d e s u b
n o r m a l g a i n s in b o d y w e i g h t a n d d e v e lo p e d m u c h le s s s e v e r e b o n e le s io n s . 30X .
42
M ICHIGAN T E C H N IC A L B U L L E T IN NO. 150
F i g . 21. C a l f C -171. S e v e r e r i c k e t s . S h o w s : s p l i t t i n g o f c a r t i l a g e , A ; o s t e o i d a n d f i b r o u s c o n
n e c t i v e t i s s u e s , B ; c r u s h i n g o f c a r t i l a g e c e ll s , C ( s e e F i g . 22 f o r h i g h e r m a g n i f i c a t i o n ) ; p a t c h e s
o f o s te o i d t i s s u e a b o u t i n d i v i d u a l c e ll s , D ( s e e F i g . 24 f o r h i g h e r m a g n i f i c a t i o n ) , 3 0 X .
F i g . 22.
A rea
C in F i g . 21. S h o w s : c r u s h i n g o f c a r t i l a g e c e ll s
r e j u v e n a te d c a r t i l a g e c e lls , B . 460X .
A-
PATHOLOGY OF RICKETS IX DAIRY CALVES
F i g . 23.
C a lf. C -171.
S e v e re ric k e ts .
S h o w s : e m b r y o n i c m a r r o w , A ; o s te o i d a n d
f ib r o u s c o n n e c t i v e t i s s u e , B ( r e f e r to B i n F i g . 2 1 ); p a t c h e s o f o s te o i d t i s s u e s u r r o u n d i n g
i n d iv i d u a l c e lls , C ; e a r l y o s s if ic a t io n in p a t c h e s of o s te o i d t i s s u e s i m i l a r to t h o s e a t C ,
D , ( n o te c h a r a c t e r i s t i c h a lo e f f e c t d e s c r i b e d in t e x t ) ; r e o r g a n i z a t i o n o f t h e lo o se c o n
n e c t i v e t i s s u e b e tw e e n t r a b e c u l a e g i v i n g r is e to e m b r y o n i c b o n e m a r r o w , E ; t r a b e c u l a
w i t h c o r e o f b o n e a n d m a r g i n o f o s te o i d t i s s u e b o r d e r e d b y o s t e o b l a s t s , F . 120X.
F i g . 24. A r e a D i n F i g . 21. S h o w s : i n d iv i d u a l c e lls s u r r o u n d e d b y c i r c u l a r
z o n e s o f o s te o i d t i s s u e w h ic h s t a i n d e e p ly a c id o p h ilic , A . 460X .
MICHIGAN T E C H N IC A L B U L L E T IN NO. 150
C 188
C 238
C 171
C 231
F i g . 25. R o e n t g e n o g r a m s ‘if m id ■f r o n t a l s e c t i o n s 4 m m . in t h i c k n e s s t a k e n f r o m t h e left,
e i g h t h c o s t o c h o n d r a l j u n c t i o n o f e a c h c a lf a s l a b e le d . T h e v a r y i n g s h a d e s o f b l a c k s h o w t h e
d i s t r i b u t i o n of c a lc i f i e d t i s s u e s .
( -151 s h o w s a n o r m a l c o n d i t i o n .
C -1 4 8 , C -155, a n d C -171
s h o w r i c k c t i c c h a n g e s a s e v id e n c e d b y d e f i c ie n t c a l c i f i c a t i o n a t t h e e n d s o f t h e b o n e s . T h e
r o e n t g e n o g r a m of ('-1 S 8 s h o w s s l i g h t e v id e n c e o f a t l e a s t t w o n r r e s t e d - g r o w t h l i n e s i n t h e
v e n t r a l e n d o f t h e d i a p h y s i s . X o t e t h a t b e a d i n g of t h e r i c k c t i c r i b s is m o r e p r o n o u n c e d o n t h e
m e d i a l s id e . A l s o , n o t e i n c r e a s e d c u r v a t u r e o f r i c k e t i c r i b s in a m e d i a l d i r e c t i o n .
F i g . 2(>. R o e n t g e n o g r a m s o f m i d - f r o n t a l s e c t i o n s 4 m m . in t h i c k n e s s t a k e n f r o m t h e l e f t
e i g h t h c o s t o c h o n d r a l j u n c t i o n o f e a c h c a lf a s l a b e l e d , f 1-224 s h o w s e v i d e n c e o f e p i p h y s e a l t y p e
of h e a l i n g r i c k e t s ( r e f e r t o T a b l e 6 ) . C -2 2 9 , ('-2 3 1 , a m i ('-2.33 a r e f r o m n o r m a l c a l v e s . C -2 3 2 ,
C -237. a n d ( -258 a r e f ro m r i c k c t i c c a l v e s .
X o t e t h a t t h e r i c k e t i c a l t e r a t i o n s i n C -2 3 7 a r e
r e l a t i v e l y m ild w h e n c o m p a r e d w i t h (1-258, b e c a u s e of p o o r g r o w t h ( r e f e r to T a b l e s 8 a n d 9,
a n d F i g s . ]9 a n d 20).
PATHOLOGY OF RICKETS IX DAIRY CALVES
F i g . 27. M i d - f r o n t a l s e c t i o n s 4 m m , in t h i c k n e s s t a k e n f r o m t h e le f t e i g h t h
c o s t o c h o n d r a l j u n c t i o n of e a c h c a lf a s la b e le d , a n d s ta i n e d in s il v e r n i t r a t e
s o lu t io n . S h o w s t y p i c a l a p p e a r a n c e o f n o r m a l a n d of r ic k e ti c r ib s w h e n t r e a t e d
in t h i s m a n n e r . C o m p a r e w i t h F i g . 26 . N o t e t h a t w h ile t h e g e n e r a l a p p e a r
a n c e s a r e t h e s a m e , t h e r e is g r e a t e r d e ta i l in t h e r o e n t g e n o g r a m s t h a n in
s p e c i m e n s s t a i n e d in s i l v e r n i t r a t e s o lu tio n .
F i u 28
R o e n t g e n o g r a m s o f m id - f r o n t a l s e c t i o n s 4 m m i n t h i c k n e s s t a k e n f ro m t h e
l e f t e i g h t h c o s to c h o n d r a l j u n c t i o n of e a c h c a lf a s la b e le d . C -176 s h o w s s o m e r a r e f a c t i o n ;
t h i s c a lf w a s f r e e f r o m r i c k e t s . C -170, C -173, a n d C - 17a a r e s p e c im e n s f ro m r i c k e t i c r ib s
I n c a s e C h l7 5 , d e f i n i t e e v id e n c e o f r i c k e t s w a s i n d i c a t e d o n ly in t h e b lo o d a n a l y s e s a n d
in h i s t o l o g i c a l s t u d i e s o f t h e b o n e .
45
46
M ICHIGAN T E C H N IC A L B U L L E T IN NO. 150
F i g . 29, C a lf C -232. S e v e r e r i c k e t s . S h o w s c h a n g e s s i m i l a r t o t h o s e in F i g . 20. C o m p a r e
w i t h F i g . 10, C a lf C -1 7 0 , s h o w i n g c o m p a r a t i v e l y s l i g h t a l t e r a t i o n s . F i g . 29 is f r o m a y o u n g e r
c a lf w h ic h h a d r i c k e t s f o r a m u c h s h o r t e r t im e t h a n d id C -1 7 0 . T h i s i l l u s t r a t e s t h e m o r e e x t e n
s iv e a lt e r a t io n s w h ic h a r e p ro d u c e d in r ic k e ts in y o u n g e r c a lv e s . 30X .
F i g . 30. C a lf C -1 8 8 . A d v a n c e d r i c k e t s in a n o l d e r c a lf .
S h o w s : c a lc ific a tio n of
m a tr ix , A ; e m b ry o n ic m a r r o w a n d c o n n e c tiv e tis s u e , B ; m o re th a n u s u a l n u m b e r of
b o n e t r a b e c u l a e , w h ic h a c c o u n t f o r t h e a r r e s t e d - g r o w t h l i n e s i n d i c a t e d in F i g ?5 C
30X
PATHOLOGY OF RICKETS IX DAIRY CALVES
F i g . 31. C a lf C-14S. R ib s
e p ip h y s e a l e n d s
of th e
b e st seen a t A .
f r o m t h e r i g h t s id e . S h o w e n l a r g e m e n t s o f t h e v e n t r a l r ib s . T h i s c o n d itio n is c o m m o n ly r e f e r r e d to a s b e a d in g , a n d is
F i g . 32. C a lf C -1S8. R o e n t g e n o g r a m s o f t h i n s p e c i m e n s o f b o n e
ta k e n th ro u g h th e
p r o x im a l e n d of t h e f e m u r .
T h e r e a r e n o a b n o r m a l i t i e s a t A . b u t n o t e t h e m o d e r a te
r a r e f a c t i o n a t B a n d t h e lo s s o f c a r t i l a g e a n d r a r e f a c t i o n a t C. S e e F ig . 33.
F i g . 33. C a lf C -188. P r o x i m a l e n d o f f e m u r o f r ic k e ti c c a lf . S h o w s : p i t t i n g o f b o n e ,
A ; d e s t r u c t i o n o f c a r t i l a g e , B ; t h i n n i n g of c a r t i l a g e , C . ( N o t e t h a t t h is s p e c im e n h a s
b e e n r e c o n s t r u c t e d t h r o u g h t h e c e n t e r a t D w h e r e a s e c tio n o f b o n e w a s s a w e d o u t f o r
h i s t o lo g i c a l s t u d y ) . E x t r e m e p i t t i n g a n d e r o s i o n o f t h e a r t i c u l a r s u r f a c e s , s u c h a s i l l u s
t r a t e d h e r e , o c c u r r e d b u t o n c e in c a lv e s u s e d in t h is s tu d y .
F i g . 34. H u m e r u s o f a r ic k e ti c c a lf . S h o w s :
a c o m m o n c o n d it i o n in r i c k c t i c c a lv e s .
p i t t i n g of a r t i c u l a r s u r f a c e A .
T h is w as