BLOOD STUDEES EN DOGS
FOLLO‘NING THE ADMiNISTRATION
OF SULFAMERAZINE

Thesis for the Degree of M. S.
MICHIGAN STATE COLLEGE
F. E. Eads
1949.

 

 

 

 

This is to certify that the a; i
. , .J'
thesis entitled . g 13
Blood Studies in Dogs following. i
the Administration of Sulfamerazine
presented by . ‘ 7 ' ‘ I
F. E. Eads §
has been accepted towards fulfillment r E
of the requirements for ' . ~ 1
it
Easter Of sciem degree 111W Medicine E
_ jab
- l
3
Major prcfuor é
' 5
Date Liav 20; 1949 i
f .
E .
1 ;
a z '
f
!
i .
l 1
:- * '
i
l
r ’ 4
-i 3
i t '
4 1' a

 

 

fi 'fi.‘ ‘1 r ‘ ulna -u- —~‘ -‘ “so -‘. (\- A‘. ’-1
LO 1) -- * ‘ . . «

.J 'L‘ y-uU-'...LJ .4.-. JUMJ

w? u" - 1“ '7‘.“ '“WY..-‘ a :I‘. c-:—- -—s- "7". {'1’... -
.. my..-‘...-J 4.9.4 £1.4J..g-..;.t.i...-..'.4...\.’-

. —p~-
‘

7 «“1 C‘ ""m
'I

J

7 an 73-. \n

-. .34. -AL'JV
.A-‘
D"

."_1‘ ‘ ‘fi “‘ "t
A LAMJJL.)

-1. ‘.,..°.;.' 1 ' 4* r‘ r. ' n 1 n -._. 1.3",
easilocoe to one eraudate ecgool o~ h_cniuen
r-‘"_ H . .“1 _o ‘ q -'.~ " "I .'. ““H1 0 j
ovate College of “or;01;o¢re age Arginog
'1 .- .- , H“- 4 .0 n. n: .. .1- n -m’
goience in {eroial Lil;;lflenb o“ uAO

. ,_ ‘1: v. QJ— 46‘ ‘ "‘1 "1 " n
re4eironetes nor -do uCUSOG 0*

$" ‘f'T 1'1 r."‘| ’_‘~‘\_‘ 1“"\'_\

.. -1-.. J. .0 '1‘ . ,.. .. - , ".-," ' :‘
egaro one or QdeCrJ and “oaucanc

1'. "1
u; 5.) J41“... -441444...u

THESiS

 

To
I"? wife, DOLLY and

daughter, IIALTCY AKIIE

216971.

‘- ”We :3” r“
Hui“. S‘s-O—J- U. ‘5.ng

.L

_L o o _ 4.1 J_V o 9 fl 0
Io is wise pleasure snot o13 writer e; resses his

most sincere ep_7recis'ion U0 Jr. C. 3. niJen for his encourage-
ment sad invaluable assistance in csrr'ing out this project.
Without his able 'rid.ance in solving many technical problels
this investig: tion U011d not have been possible.

Gratitude is also overesr: ed to Drs‘ .H. J. Staiseth,
L.3. Sholl, and Frank Th0°p, Jr. for many v0 liable susgestions

during the course of tnis s iey and their contributions on the

final arrangement of this manuscript.
Sincere sppreci wtion is ale 0 expressed to Dr. S. F.

Scheidy, 3h erp and Dohne, Inc., Glcnolden, Penn"slvnnie for furn-

all of the snlfa:erszine used in this study.

(1
Cl
*4.
(a)
(‘3'
a?
g .
P.
{:3

Tharks are due to many others who have a‘

vari 03.3 ways.

pm”

‘L‘v.....5

'T‘l“. ‘\““1 C‘fi “: 1 1V "'1/‘1
.L

lALJAJJ

<1-
vi-‘J 000000.000

uloegxen

q
I

‘I‘

.. :8 U:;OCLb)

\" ' - r3
‘1 p4.) v

-L

5-) Un—asL‘a‘Uu— A.

eterninati_on of none; 10bin

:ion 00.0.00...

.
’5:“:"‘
A ~U‘ ”vile 00.

!V\\( 4"“ ~a of :3 T I‘m-u.

—II-"fl L".~u

3\fis.~uJ-I-L.J

OOCCOOOOOOOOOOOOOOOOOO0.0......

fi“f‘r"§"’\ 4* (‘3 -
.v-L ”’I‘J -.—'v p—al Ix} n-n-I‘C'

-. ‘. — . *1. J—‘ 1* .
L)LL-OCJPCO {ll-:0. .-_'_4.‘.1u‘ LILLL ”00‘; W3 “11.71.31 5‘ .3. \J1
:09. . -. ° 1- - i- _-.g-, n . “.5
-1ier.atiel neinoeJoe oosnos ....

Oral Ai:inis ration

a 1 '_ . -3 L rL: H

.41.. .10C Ll' U841 601.18 $135.11... 4'1 -LS Urfi. ULOJ'. O O O o o o
+ . M ._

:lu filv'el'l 4.3 £151.. 1-11.13 U15. " b-Oll . o o o o 0

“fl". ‘

QOQLOH 000000000000.000000000000000
v.19 H . ~ e1“-.: A -1 r. of ~
.3 its. . "Mordzine Au...s...1‘-'—S U0 ‘pCL .LJC. CL 5).}. 1
C. ». f5 ‘ a O a 1-.— _ fi" 1
alliancrsZine new nisbel “ed; . e.ere.

n6

0
-1 <_—

—-‘n-

w-..-

-011

.‘13‘

D
-V-f

uOlJ 3;»:

O
rm -,- “5—1
UV“

0.00.0000...OOOOOOOOOOOOOOOOOOO

11.]. 00.0.0...0.0.0...OOOOOOOOOOOOOOOOOOOC.0...

OOOOOCOOOCOO00.000.00.00..0OOOOOOOOOOOOIOOOOO.

:Jo
f—I [>40 '40

{35

t3

9?

‘VN

'3’")
£51.

’jfi

A)

-‘ IIG

‘- ....._

11

0 1‘0
Kn.

I
O
‘I

- a
‘1

LAn-J
(l...

‘
'c3 '1

U

u .
.'"‘.‘3
«54v

‘0!
J.
O

o
C
F"!

."\h
.L

o
.2.
U
N. 1°
-.-5~...n

are

rat
ca
)

.
L
1
\l
‘\f"
«LJ
0
(I
“7

blood

V

G;

g

no;

'I

\J sz-L _.-‘L&.'..s J
s~r~

o¢>J

\LLLLCLL

O

WY‘.*
0

7"“ PI
AAA...)

0
a

1
.3.
C
.L
" uO
‘1
.u,

~

A;

we v0
Lie

v.-
(ILL-
U 5......
TO 1
Q

I
’5
rwfl'l
.—-~L__ vb U...
l
fi"'r‘

.‘3’?
-.~---.L....\.~v

‘ L

n

5"

I"

4"

.1.

o

‘w ‘1‘
f

...- .U
‘0

{ TWV“
Co

a-.
C

3fi'?”
--u
0 A
.1.
no C
J-
.
.9
1r‘
chi.)
.
'5‘qu '1 7a
-.

l .

(«wfir‘v
‘4—

‘A'! O
5)

(“1"
V--... -
n
(«.1 -
D v-_.'~.‘.
J.
‘3“
L41. LI
’3
‘3
nit—n
(.1L- V -.
1 v7
.. .LCV
I‘V‘
~_-..A.4.0.L L.

+..

C

1
I1

01"

r‘ 7'" n
n

Thv
-U...
‘—'..v-.-54‘
In”
'VJ-
‘-
.L
‘L 51’\_ I
U.- V“ U-.-
0
7.9!} '7
It .L~o.—‘
J...
V
fl
“5)-.--
.N]

O
:3

'\""1h
‘_'1
.‘,
150.
cav-
o o
21.0 1.
“’N
-15.
'0 o
'" '31
5gb
1111
o
o
't V.‘
U.LV
5“
i)

K“

r'
O
.
u.lL
V
'-
0 Oil

”—1
I")
“U...

.J‘I‘O

ec
n t
5)
fo
.r
c

it o
J
fif‘-
1

A-

C
1.
L}

J.

T"f‘1"~f\
.—.‘. .5-o
6“".

“J.

.

‘3 .1
L14. .— _,

O

O
a

0
71", ‘

1

f

a.‘

o

’3

“AA—5.}
“4.-“V

3.3
-r

J

-.¢“U
I
‘N
3.05.
‘\
' I
s-’
1
O
’V 1‘

U

OTC
d:
54.1.1.

LA.

..4
.
H

.118
yr.
l
A“ -
r}
V 1 o"

l

..
n'nn

by...“
’37.

A La
."o‘v
e“

0.1

N: "8’flf1
all...v...~ ,

W

I a. _

2

‘
o

C)

:17.

sa '1' p
0.- u .L 01‘
I‘ICZ‘G‘

One
5“

O

O
_‘

1
.e
’.‘
6—!
a.
>- 1‘
L'...C.LA

‘;b

10; of

I‘Cl

C 01170 I‘.

.J
)
hid

.

LOHC l‘
V

€11.“ "‘

3;

~

(331

011.10

t

I
..
.L
.
.,
O”
.L
1.0
--\
‘

~ <

.0
I

.»V V'
¢.-
.—

ru- 1 f. r. -“-~.‘
0

v
f?“
95‘.
15

('3 T“.
A.

pract
(y
on
th
or
he

‘

g
\ r:)
li‘.’

.Tu

‘\ O 0 l
-‘I ’1‘ _-
.. 3) V

3/51? LLD

4-"
u.)

.1

REV I IN OF LI T’ER’JI'URE

The results obtained from blood studies following
the administration of sulfamerazine in various animal species
have been reported. Limited experimental data comparing the re-
sults obtained by the administration of sulfamerazine in dogs by
various routes and dosages have been published.

Sulfanerazine (2-sulfanilamido—Admethylpyrimidine) is

represented by the following structural formula.

N=CH

"N--1 CH

O H N C---CH3

Sulfamerazine

Sulfamerazine has distinct advantages over other sulfo-
namides in veterinary practice in that it is more rapidly and completely
absorbed from the gastrointestinal tract and more slowly eliminated
in the kidneys (Nelch et al., 1943), (Goodwin et al., 1942), (Murphy
et a1., 1943), (Bryan, 1946) (Francis, 1947), (Hastrack and Lewis,
1947), and (Soheidy and Tillson, 1947). Therefore, therapeutic blood
levels are maintained for longer periods and these levels can be pro-
duced and maintained by smaller and less frequent doses of the drug.
In.man, sulfamerazine is mainly absorbed from the small intestine
and only slightly from the stomach with the large intestine inter-

mediate in absorptive capacity'(Hurphy et a1., 1943), and (Welch et
a1., 1943).

-2-

Sodium bicarbonate administered by mouth in sufficient
amounts to increase the urinary pH from 6.8 to 7.8 increased the
clearance of sulfamerazine two or three fold (Peters, Beyer, and
Patch, 1944). This was likely due for the most part to increased
electrolyte excretion rather than to pH change. Beyer et al., (1944)
reported that the administration of sodium bicarbonate increased
the urinary pH but it also increased the clearance value of sulfa-
merazine by interfering with its reabsorption. As the plasma level
of the compound was increased, and more of the drug was filtered by
the renal glomeruli, there was an increase in the amount of the
sulfonamide that was reabsorbed through the renal tubules; there-
fore the ratio between plasma concentration and the amount excreted
per unit of time remained fairly constant. Earle (1944) found an
observable decrease in the reabsorption of sulfamerazine following
the administration of sodium bicarbonate. Peters et al., (1944)
reported that the clearance of sulfamerazine remained fairly const-
ant when the plasma level was increased from 3.4 to 13.1 mg. percent
indicating that maximum tubular reabsorption of the compound was not
exceeded. Assuming complete glomerular filtration of unbound sulfa-
merazine, normally about 80 percent of the filtered compound is re-
absorbed by the renal tubules of the dog. Similar results were re—
ported in man by Reinhold et al., (1945). Earle (1944) calculated
that in man 63 percent of the sulfamerazine is bound at a plasma
concentration of 10 mg. percent. There is a more extensive binding
of sulfamerazine in human than in dog plasma, and the renal clear-
ance of sulfamerazine in man is considerably lower than in the dog.
Davis (1943) has reported that essentially all the binding of sulfon—

amides is on the albumin fraction of plasma. Beyer et al., (1946)

found that the binding of these same agents varies from one species
to another.

Welch and his co—workers (1943) found that Sulfamera—
zine and its acetyl derivatives were approximately 20 percent more
soluble than the respective forms of sulfadiazine in both water and
urine.

Sulfamerazine blood plasm concentrations have been
studied in dogs, cats, horses, cattle, sheep, swine, foxes, mink,
poultry and man. welsh and his associates (1946) have demonstra-
ted that, following the administration of a single oral dose of
sulfamerazine (5 grain per pound, or 71.5 mg. per kilogram of body
weight), it is possible to obtain and maintain higher blood concen-
trations in the dog than with sulfanilamide, sulfapyridine, sulfa-
thiazole, sulfadiazine, sulfamethazine, or sulfaguanidine. Sulfa—
merazine also gave effective blood levels in cats but did not pro-
duce adequate blood concentrations in horses, cattle, sheep or
swine. They concluded than an adequate dosage for animals is a
maintenance blood level above 5 mg. percent, and preferably above
8 mg. percent twentyhfour hours a day. Inadequate levels and fluc-
tuating blood levels are responsible for the develOpment of drug-
fast strains of bacteria. The blood plasma level studies of Welsh
et al., (1946) using-% grain (71.5 mg. per kilogram) per pound of
body weight are comparable to the data reported herein.

A similar study was made by Scheidy and Tillson (1947)
comparing the blood concentrations of sulfadiazine, sulfamerazine,

and sulfamethazine in cattle. Therapeutic blood levels of sulfamera-

zine were maintained for twenty-four hours by a single oral dose of

I
-‘+-

1 grain per pound of body weight. Intravenously, using-g grain per
pound of body weight in cattle, similar results were obtained to
those shown in Table XXIII.

In a more recent study on blood concentrations follow-
ing the administration of sulfamerazine and sulfamethazine to cattle,
Scheidy et al., (1949) found that only a very small percentage of the
absorbed drug is acetylated by cattle. A combination of these two
drugs caused no apparent serious toxic reactions so far as could be
determined, and the values for creatinine, non-protein nitrogen,
icterus index, hemoglobin and average blood values appeared to be
within the normal range during each phase.

Welsh et al., (1948) in studies of sulfonamide blood
concentrations in foxes found very low concentrations of sulfamera-
zine when-é grain per pound was administered in a single dose per 05.
Similar studies were made by Langer et al., (1948) in minx. Much
higher blood concentrations were f and, in fact,-§ grain per pound
in a single dose per 03 gave a therapeutic blood level for eight
hours and compared favorably with sulfamethazine and sulfadiazine.

Sulfamerazine blood levels in poultry have been studied
by Thorp et al., (1947), Gordeuk and Learned (1948) Kiser et al.,
(1948), Nattis et al., (1946), Severens et al., (1945), Bottorff and
Kiser (1947), Anderson et al., (1947), Alberts and Graham (1948),
and others. Thorp et al., (1947) observed that sodium sulfamerazine
administered in a concentration of 0.2 percent in water for five days
resulted in an average blood concentration of 13.68 mg. of free sulfa-
merazine per 100 ml. When 1.0 percent sulfamerazine was fed in the

mash, an average blood level of 27.08 mg. of free drug per 100 ml.

[-0

of blood resulted. Comparable results were obtained with 0.25 per-
cent concentration of sulfamerazine in the mash. An average of 8.26
mg. of free drug per 100 ml. of blood was obtained during a five-day
treatment. Gordeuk and Learned (1948) and Kiser et al., (1948) re-
ported similar results in chickens and turkey poults. Alberts and
Graham (1948) obtained blood sulfonamide concentrations of 16.4 mg.
percent within three hours after the fourth oral dose of sodium sulfa-
merazine (12 hour intervals, 0.5 grains per pound) was administered to
turkeys. Using 0.2 percent sodium sulfamerazine in chickens, Anderson
et al., (1947) obtained blood concentrations of 16 mg. percent.

Mattis et al., (1946) obtained the same blood concentration when the
dosage of sulfamerazine in the feed was increased to 0.5 percent.
Blood concentrations of 30 mg. percent or higher were obtained by
Severens et al., (1945) with 0.5 to 2.0 percent sulfamerazine. IBottorff
and Kiser (1947) reported blood concentrations ranging between 12 and
31 mg. percent when 0.5 to 0.75 percent were administered.

'xtensive work has been done in humans with the various
sulfonamides. Gilligan (1943) in his studies on the plasma binding
properties of sulfadiazine, sulfamerazine, and sulfametnazine in man
found that plasma levels of sulfamerazine were considerably higher
than for sulfadiazine suggesting that this drug is bound in the plasma
to a greater extent. As the plasma concentration decreased, the per-

n vitro dialysis studies

cent of binding of the drug increased.

 

show sulfamerazine to be bound in the plasma to a greater extent than
sulfadiazine. The binding of the drug in the plasma ip vivo was essen-
tially the same as was found in the ip vitro studies. Sieber and

Clark (1945) stated that plasma binding alone cannot account for pro-

longed blood levels of sulfonamides. Other factors which determine

the magnitude and maintenance of blood levels are: 1) solubility in
and absorption from the intestinal tract; 2) presence or absence of
food; 3) the emptying time of the gastro-intestinal tract; 4) ex-

cretion into the intestinal tract; and 5) urinary and fecal excre-

tion.

Welch and his associates (1944) found that when an
initial 2 gram dose of sulfamerazine, followed by daily maintenance
doses of 1 gram was administered, the average daily concentration of
free sulfamerazine in the blood of 8 normal men decreased from about
6.5 mg. to 3.5 mg. percent. Hageman et al., (1943) gave an initial
dose of 4 grams by mouth and a maintenance dose of 1 gram every 8
hours and obtained blood plasma levels of 20 mg. percent at two hours
which decreased to 14 mg. percent in five hours.

Murphy and his co—workers (1943) reported that when
. sulfamerazine was administered rectally in aqueous suspension by re-
tention enema, at no time was more than a trace of the drug found in
the blood.

Renal elimination and toxicity are the two primary
concerns in sulfonamide therapy. Important toxic manifestations are
granulocytopenia, hemolytic anemia, and kidney concrements (Green,
1944). The low toxicity of sulfamerazine has made it the drug of
choice in the treatment of systemic bacterial infections. The lower
concentration in the urine and the fact that both the drug itself and
its acetyl derivative were more soluble in neutral and acid urine
than the corresponding forms of the older sulfonamides,lessen the

possibility of renal obstruction (welch et al., 1943). Welch and

-7-

his associates also reported that the toxicity apparent -y was about
the same for sulfamerazine and sulfadiazine when a comparison was
made on the basis of the concentration of the compounds in the blood.

Schmidt et al., (1944) compared the toxicity of sulfa-
merazine, sulfamethazine and sulfadiazine. Insofar as acute and/or
chronic toxicity are concerned there appeared to be little difference
between the three compounds. However, both sulfamerazine and sulfa-
methazine seem to be more desirable than sulfadiazine as far as solue
bility and renal toxicity are concerned.

Studies by Leh r (1945) and Fri isk et al., (1947) with
various sulfonamide compounds have shown that there is a reduced
chance of renal toxicity in small ez-zperimental animals and in humans

.
tn

when rixtures of sulfonamides, ra . 1am one or the other of the
components, are given.
Mattis et al., (1946) conducted toxicity studies with
young chickens and administered 0.5 and 1.0 percent sulfamerazine
in dry meal. for l4s1ccess ive days. This dosage of the compound pro-
duced no evidence of toxicity in the chiclzoxm Average concentrations

7
|

of free sulfamerasine in the blood of the treated cuic} :ens we re re-
spectively 14.7 and 29.1 mg. percent when the drug was fed in these
concentrations. Farr and Jaquette (1947) reported tox;ci.ty in cl1ick—

ens with sulfamerazine in the mash at levels higher than 0.25 percent

or a week.

0
L)
L ‘3
m.
L;-
i .J
(9
{I}
H)

or in 0.5 gram daily doses administered in
They observed a retarding of weight gain which they felt was due
both to the unpalatability and to the toxic effects of the drug.

Hall ands Mi {(1943 ) reported a low incidence of toxic c-

ity in 113 human cases. Leukopenia with or without granulocyt aenia

occurred in about one percent of the cases. Similar results were
reported by Green (1944), and Clark and his associates (1943).
Kracke (1947) stated that the sulfonamide drugs, in addition to
causing leukopenia, may produce a marked degree of neutrophilic
leunocytosis.

Schmidt et al., (1944) conducted toxicity tests on
mice and found that animals receiving lethal doses of sulfamerazine
died within 18 to 36 hours after treatment. When 5 grams per kilo-
gram of body weight was administered by mouth a blood concentration
of 148 mg. percent was obtained. This killed 93 percent of the nice.
When a 2 gram per kilogram dose was administered subcutaneously, a
blood concentration of 164 m5. percent was obtained which produced a
90 percent mortality. When the same dosage was administered intra-
peritoneally, a blood concentration of 161 mg. percent was obtained
which was 95 percent fatal. They found the toxicity of sulfamerazine
for the urinary tract to be evident when blood levels exceeded 40 m5.
percent. In animals with such blood levels, ureters, bladders and
kidney pelvis frequently contained sulfamerazine concretions composed
of both free and conjugated drug. The incidence of these deposits
rose progressively with the blood levels of the drug. Sulfamerazine
seemed to have somewhat less urinary tract toxicity in the dog than
in the rat (Schmidt et al., 1944).

Beds (1949) reported that sulfamerazine may be admin-
istered safely to dogs and cats in daily dosages of 1 grain per pound
of body weight in divided doses. In this clinical study no visible
untoward effects were noticed after 7 to 11 successive d‘1s of treat-

ment.

-9-

Leukopenia following the administration of sulfanera—
zine occurs in man but not in experimental animals (Litchfield,
1948). Data reported herein show that a marked leukopenia developed
upon the administration of large doses of sulfamerazine to normal
healthy dogs. Clark et al., (1943) observed leukopenia in man with
sulfamerazine therapy. They reported no cases of acute hemolytic
anemia or agranulocytosis. Jennings et al., (1937) reported a few
cases of agranulocytosis, all with fatal termination. ho agranulo-
cytosis or granulopenia occurred even when a leukopenia developed
during sulfonamide therapy (Bigler et al., 1938). The action appeared
to be independent of the leukocytes in that it did not produce an in-
crease in total leukocytes or in the proportion of neutrOphiles.

According to Denny and henten (1946), blood dyscrasias
are among the less frequentl1 observed pathologic changes produced by
the sulfon mides which include hemolytic anemia, aplastic anemia, and
agranulocytopenia. Aplastic anemia was rare in both children and adults
but hemolytic anemia was seen rather frequently. Favorite, Rainer and
London (1944) reported a case of leukOpenia in man due to sulfamera-
zine when 42 grams was administered during a 23 day period. No changes
in the red cell count or hemoglobin content were noticed but a marked
lymphocytosis (93 percent) and neutropenia (7 percent) was observed.
Menten and Graff (1946) conducted a survey of the hemoglobin and red
cell counts of children whose blood exhibited granulocytOpenia and
found that they did not usually reveal an accompanying anemia. The
percentage of neutropenias developing from sulfonamides in their stud-

ies was about 20 percent of all patients receiving the drug.

Malkamus and Opperman (1944) reported the normal ranges

-10-

of blood constituents in the do; as follows: erythrocytes 5.5 to 8
million per cu. mm., leukocytes 9 to 10 thousand per cu. mm., and

the leikocyte differez1tia1 coznt was 60 to 80 percent neutrophilcs,
2 to 4 percent eosinophiles, O to 0.5 percent basophiles, 13 to 32

percent lymphocytes, and 3 to 5 percent monocytes.

U1

The no mal ra13es of ti 1e canine blood constituent
reported by Coffin (1945) are as follows: er;rt11. oeytes 5.5 to 0.6
millions per cu. mm., leukocytes 5 to 20 thousand per cu. mm., hemo-
1

3lobin 12.5 to 17.3 grams per 100 ml., neutrophiles 3.6 CO 15 thous-

and er cu. m”. eosino p--ilc sO.1 to 2 thousand per cu. Hm. baso-
’ .. 2

phile 30 to 0.4 thousand per cu. mm., lymphocytes 0.6 to 6 thousand
per cu. mm., monocytes 0.1 to 2.4 thousand cu. mm.

Boddie (1946) reported the following as normal ranges
in dogs: erythrocytes 5.1 to 7.6 mi_lion per cu. mn., 1eu3zocytes 8
to L .6 thousand per cu. mm., homo: 3lobi n 11.2 to 14.363rams per 100
m1., and the leukocyte differ mitial count was 0 to 4 percent eosino-

philes, 67 to Cl percer mt neu.troz>hiles, 14.6 to 25.4 percent lfi ipho-

cytes, l to 7 percent monocytes.

-11-

LEMIIODS

Estimation g: Sulfamerazine ig the Circulatory System.

The method employed by Hoffman (l94l) was used. The concentration

of free sulfamerazine in the blood was determined by the method of
Bratton and Marshall (1939). The technique was as follows: Collect
two or three drops of blood from the radial vein in a small tube con-
taining a crystal or two of potassium oxalate. Transfer 0.1 ml. of
this to a small test tube containing exactly 5 ml. of distilled water.
The pipette should be washed out by sucking up the water and blowing
it out several times. Add 1.0 ml. of 15 percent trichloroacetic acid.
Mix thoroughly with the pipette, and again wash the pipette by suck-
ing up the fluid and blowing it back. Centifuge the sample until
the precipitate is completely separated from the filtrate. Transfer
4 ml. of the filtrate into a 10 ml. graduated tube; add 0.5 ml. of
0.1 percent sodium nitrite and mix. After 3 minutes, add 0.5 ml. of
ammonium sulfamate. After 1 minute, add 0.5 ml. of N (l—naphthyl)
ethylenediamine dihydrochloride. Let the color develop for at least
1 minute, dilute to 8 ml., mix, and compare readings in the Cenco-
Sheard-Sanford Photelometer using the green filter with a standard
stock solution of sulfamerazine. Preparation of this stock solut-
ion is as follows; Weigh out accurately 100 mg. of pure sulfamera-

zine* and transfer to a liter volumetric flask with a little distilled

water. Add a few drops of 10 percent sodium hydroxide to hasten

* Sharp and Dohme Sulfamerazine Chemical Reagent (Powder).

-12-

the solution of the comparatively insoluble powder, dilute with
distilled water to the 1000 ml. mark and shake until all the sulfa-
merazine is dissolved.

Calculation is made by reference to a calibration
curve obtained by analysis of the stock solution. This calibration
curve is determined as follows: Transfer 2.0, 1.5, 1.0, 0.5, and 0
ml. respectively of this stock solution to 50 ml. volumetric flasks.
To each of these add 9 ml. of 15 percent trichloroacetic acid. Make
up each solution to 50 ml. with distilled water, and mix thoroughly.
These solutions represent 1 to 40 filtrates of blood specimens con-
taining respectively 16, 12, 8, 4, and 0 mg. per 100 ml. Analyze
10 ml. of each of these solutions exactly as above for blood filtra-
tes, making up finally to 20 ml., and read in the Photelometer,
using the green filter. Plot the values on semi-logarithmic paper.
They fall in a straight line in the lower concentrations but may
have a slight tendency to form a convex curve in the region of 16 mg.
per 100 ml.

Determination.gf Hemoglobin. The Photelometer was
used in determining hemoglobin (Sanford et al., 1933). Preparation
of the sample of blood was as follows: Twenty ml. of the diluting
fluid (0.1 percent solution of sodium carbonate in water) was meas-
ured accurately into a suitable container such as 50 ml. Erlenmeyer
flask. To this was added 0.1 ml. of blood. The mixture was thor-
oughly shaken and then transferred to the absorption cells and
checked in the Photelometer, using the green filter. This reading
was then obtained in grams of hemoglobin per 100 ml. of blood from

a calculated table.

-13-

Leukocyte and Erythrocyte Enumeration. Leukocyte
and erythrocyte counts were made within 24 hours after bleeding
using Thoma diluting pipettes and Bright-Line Improved Neubauer
Counting Chambers (Coffin, 1945). Two percent oxalic acid (Jones,
1927) was used as the diluting fluid for counting leukocytes, while
Leaks and Guy diluting fluid (Todd, 1943) was used for counting
erythrocytes.

Differential Leukocyte gggptg. Blood smears were pre-
pared within one hour after bleeding by the two-slide method using
oxalated blood and stained with‘Wright's stain (Todd, 1943) immedi-
ately following drying.

The dogs used in this study were obtained from a city
dog pound. All dogs were hospitalized for several months prior to
use. None of the dogs developed distemper or any other contagious
disease throughout the course of study. The ages ranged from app-
roximately fifteen months to five years. The animals were fed once
each day; the ration consisted of one part of cooked meat scraps
and two parts of commercial cereal dog food. This was fed at approx-
imately three p.m. each day. 'water was kept before the dogs at all
times. Sulfamerazine administration and the drawing of blood samples
were always begun in the morning, and blood samples were collected
at regular intervals until that phase of the experiment was concluded.
All of the animals gained weight steadily during the entire course of
study. '

The anti-coagulant used in collecting blood samples

was 2 percent potassium oxalate*; one-half ml. of this preparation

* l Gm. of potassium.oxa1ate was placed in 50 ml. of distilled water.

-14.

was placed in each bleeding vial and placed in a paraffin oven
until dried. Each vial therefore contained 10 mg. of potassium
oxalate which is enough oxalate to prevent the clotting of 5 ml.
of blood.
The types of sulfamerazine products and routes of ad-
ministration used in this experiment were: 1) in tablet form, 2)
in powder form, 3) in an emulsion, 4) intravenously, and 5) subcut-
aneously. Each type of product was administered at the rate of-%
grain (71.5 mg. per kilogram), 1 grain (143 mg. per kilogram), and
-% grains (214.5 mg. per kilogram) of sulfamerazine per pound of body
weight respectively. Further blood plasma concentrations were also
determined when sulfamerazine was administered once, twice and three
times daily for a three day period as compared to a 24 hour period.
The first objective in this study was to compare sulfa-
merazine blood plasma levels following the single oral administration
of-% grain (71.5 mg. per kilogram), 1 grain (143 mg. per kilogram),
and 1% grains (214.5 mg. per kilogram) per pound of body weight.
Dogs numbered 101, 102, 103, 104, and 105 were used. Tables IX
through XIII. In this series of trials dogs were weighed immediately
prior to administering the drug in tablet form. One hour following
the administration of sulfamerazine, the first blood sample (approx-
imately 1 ml.) was drawn from the right radial vein. Blood samples
were then taken at 4 hour intervals for the next twelve hours fol-
lowed by another at twenty-four hours. Beginning with the first
sample drawn, blood was taken at specified intervals and checked for
red and white cell counts, hemoglobin content, and leukocyte differ-

ential counts, to determine if sulfamerazine had any effect on alter-

-15-

ing these in the normal healthy animal. Similar series of trials
were conducted on dogs numbered 115, 116, and 117 (Tables XIV through
XVI) using the powdered sulfamerazine given by mouth in capsules,
and on dogs numbered 118, 119, and 120 (Tables XVII through XIX)
using an emulsion measured in a 10 ml. syringe and deposited in the
mouth to assure proper dosage. In these trials, blood samples, red
and white cell counts, hemoglobin determinations and leukocyte dif—
ferential counts were studied and compared with the results when the
tablet form of the drug was used.

In the next series of trials a sterile aqueous solut-
ion containing 6 percent sulfamerazine was used on dogs numbered 121,
122, and 123 (Tables XX through XXII). Subcutaneous injections
were made in the nuchal region approximately on the midline. This
location was chosen as it is readily accessible and caused the least
discomfort to the animal. A 20 gauge needle was used. Following
the subcutaneous injections blood samples were drawn one hour follow-
ing the injection, and then at 3 hours, 5 hours, 8 hours, 12 hours,
24, 36, and 48 hours. In this series of studies samples were taken
at the beginning and at twentybfour hours of each individual trial
to determine the effects of sulfamerazine on the blood picture when
given by this route.

Intravenous injections were made in the right radial
veins of dogs numbered 124, 125, and 126 (Tables XXIII through XXV)
using a 20 gauge needle. The first blood sample was drawn immediately
following injection before the needle was removed from the vein,

using a clean syringe to draw the blood sample. Another blood sample

was drawn one hour after injection and thereafter at two-hour inter-

I

~10-

vals for eight hours, then at 12, 24, 36, and 48 hours. As with
the other routes of administration, the blood picture was deter-
mined at specific intervals.

Subsequent trials were conducted on dogs numbered
106, 109, 112, 107, 110, 113, 108, 111, and 114 (Tables XXVI
through XXXIV) when sulfamerazine was administered in tablet form
by mouth once, twice, and three times daily for a three day period.

The results of Tables IX through XXXIV are summarized

in Tables I through VIII.

-17-

 

 

 

 

 

 

Com Nam 50H QN
m.o m.q H.N NH
0.0 o.o m.m m
0.0 m.© o.m Q
o.e N.N m.a H
ma H .m
coupmnpmflaaaea
emwmpmaqaaes unwams_aeon .nfl\.we ampma mason

 

 

.Nwwsodmw cw
NHH .ooH .eoa .mOH .eoa .moH .NOH .HQH .oa mmoa .swoa panama as was“

Immmamaadm mo mmop mammflm w mo Goapmmpmfinflapd ammo map mquOHHom mam>oq mammam vooam ommho>¢ .H oanmw

 

 

 

 

 

 

 

 

 

o.m mé o.m INN
mp m.m m.m NH
H.b N.© N.¢ w
mg. p6 miw a.
m.m m.m N.H H
m .ma m .m: w” .mE
.
do
1
.
mm H m...
deflpmppmflmflapm
UmmmpmflcflEpm pamwmz hwon .QH\.ho momma mhdom
.xwpcmmmw Ga baa .pHH .mHH .oz amen .amom howsom GM mafia

Iwmoammasm mo mmov mamsflm w mo coflpmmpmflsflmm¢ ammo 0gp mmfizoafiom mHo>oq mammam wooam swamoba .HH manwa

 

 

 

 

 

 

 

m6 mé m; «N
0.0 m.< o.m NH
m.e o.m o.a m
mgr m6 min q
m.¢ m.N 0.0 H
u .ma m .mfi m .wa
.wH H m...
QOHsmppmflcflamm

eonmpmagaaea pnmflo: aeop .ma\.nm

 

popms masom

 

.pruodmw cm ONH .mHH «mHH .oz mmom

Immomsmadm Mo owed mamcflm w mo soapsppmfimflfiwd ammo map mmwSOHHom mam>mq mamwam wooam mmmmo>¢

.mowmadam as am mafia

 

.HHH mHndH

 

 

 

 

 

 

m.m O.H ms
ad ed on
m.OH o.m w.o gm
0.0H H.MH 0.H NH
m.®H m.mH H.N m
,0sz TS in m
b.mm o.mH m.mH m
m.ma 0.0H m.OH H
n .man mm .WQH m .mq
”H H .,....
deflpmnpmflcfiuda
dommwmwmflamd pznwm: Avon .nH\.nw magma mason

 

 

.afidmomdm CH mma .mNH .HNH .om mmom .hammoomspSoQSm mafia

Immoemmadm mo omop oncwm a mo mowpsppmflcwmpd exp MQHBOHHoh mHm>mq mammam pooam mmmmm>¢ .b mammu

 

 

 

 

 

 

 

o.m +\.H mi»
0.0 0.m mm
m.OH ¢.q «.H gm
m.¢H 0.0 N.m NH
0.0H 0.0 0.0 m
b1» 9
XS «.0 m
o.m v
m.0H H.HH m
m.m N
H.0N m.NH 0.0 H
H.bm m.HN 0.0H o
m .m& m .ms R .mE
MH H .m
mmmopmwmflawm psmwoz_h©03 .£H\.ho SOHpmmpchHamm
momma manom

 

 

.wHeemmge nH omH .mmH .«mH .ou mmoa

.hHmmomo>mppmH QQHN

Imamasdem mo omop onmHm a mo mOHpmmpmwzwids map mmHBOHHom mHobon mm ”Hm ©00Hm ommmoba .> meme

 

h'fi‘i-v‘rmr‘
_~—A~\J-—d-5J
w

r11 14:- J-.° .0 3:3-.2‘ ”31..
3.1.10 OUJ‘VCLLJIQ 0.1. 01‘... 9 L314

.2- L - ‘3 - I - :- ..' N :-
z_1e clood plas a _evel olrvos iollowing u
. n. .. - .° :-:-‘ 4—“ ..J '1 , : :v -. .. 1 32' .-

salianeraslne wits one plasma levels obtaixou followin coca intra-

vep nOW‘S n‘nd n‘lbn~t"‘r‘~~.xovn :n'mx 3':- ~
w. u)» chLIL'r-‘~.: Ln.) Juddbc 4.01..

'5... Ffi""'“""“"ln"fi""‘v
:‘HJJ ILJ“-J—- .1...) -aul&;bol

—-.- r. I . Vlfi O . I D < f‘ .\-n ‘. . C -. \‘
”ith a single oral administracion oi saliameraZLne in

test blood plasma concentration was btaized in

h a ‘ o ..

lou. hours. Eden-g grain (71.5 mg. per kilogr a*1) per pound of ooay
: " " .

weight was administered a concentration 01 3.4 ”

-.Q.

A“ - L v. n 1‘ r
percent Las lounu.
-. 1,. . , .fi 1 . ,1 “-3 .. 3.,
two nears lollowei oy a LpauLLll

m“ '1. ‘. , h ‘ - -\ ~. -L .p ..r‘
This concentra'ion was maintained ior

9 3-. . _,. A 3:. .13.: , 1.. I- n 1 -
all so that 2. r, porc3nt was So_ll present at cLentj-iour no:“s.

’Mi‘h 1 grain (143.5 mg. per kilogram) per pound of body weight, a

blood pla3na concentrac on of 6.5 mg. porosnt was obtained at 4 h urs,
with 5.1 :3. still 0:3sont at S h Hrs; at tw3nty-four hours the con-
centration of 3.2 m3. percent. Because of this remaini'ilg blood plasma

conc:3 ntrat tion, a forty-eight hour sample Las dra wn which contained

. - 1 - , __ - .. - 3
0.8 mg. percent. USing l- 'rains (2 +.5 m' per kilo ram) per pound

a. U L).

n+9

of body we i"11t a concanorzt ion of 0.4 m" sereant was obtained a3 ozr

heirs, with 5.5 r” p0“ 100 ml. at twelve hears and 5.0 mg. still pres—

.00.

r‘

cut at twenty-four heirs. compara ole results were obtaino usixg

v. o o ‘ _ ‘ fl . '1 “A ‘1'"... .
s Llfanerasize in powder for; as well as an enlision.
fl""‘~:""f‘1'." ‘f-J'j l".' ’-.“'1‘ ‘.'1 ‘I ""I"‘I\‘ T
AJNJ_4 H'dfl‘l‘l-“Jk Um) Ayn-uo-u—J-u-Ld‘A-u-INK‘I

 

r"? 1 1 ' .1 1w ; '5‘ . ‘ s-‘\, 1 *5" ~' -<
1.10 31006. b-3813. 1.0 f8 ..3 ~.3 63.11.03. c..." v33." S-.'.UC 19-1100 «4-3

10

a L an ‘ r‘ 'L r-q n3 11¢ '9 ‘n -1v ‘ v - ~ "V 1*?
auu_ni3.:ac on oi the 3311133- size co pai3d very closely wi h the

-93-

intravenous injection both in regard to the highest concentration
obtained with each dosage and to the rapid drop in concentration
from this point to the forty-eight hour readings. The highest con-
centration of sulfamerazine, when given by this route, required
approximately three hours after which time it was rapidly eliminated
from the circulatory system. The 24 hour samples contained 0.2, 8.0,
and 10.5 mg. percent of free sulfamerazine respectively when-%, l,
and If grains per pound of body weight of the drug were injected.
The 36 hour samples had 2.4 and 8.7 mg. percent while the 48 hour
samples yielded blood plasma readings of 1.0 and 5.8 mg. percent,
respectively, of sulfamerazine when 1 and 1% grains per pound was the

dose.
INTRAVENOUS ADXIEISTRATION

Since the sulfamerazine was introduced directly into
the circulatory system when given intravenously it was found almost
immediately at the concentration of 13.0 mg. percent with-§ grain per
pound of body weight, 21.3 mg. percent with 1 grain per pound of
body weight and 37.1 mg. percent with 1% grains per pound of body
weight when injected in a single dose. The concentrations fell quite
rapidly so that at twenty-four hours blood plasma levels of 1.4, 4.4,
and 10.5 mg. percent respectively were obtained. Since rather high
blood levels were still present at twenty-four hours when 1 grain
and 1% grains per pound of body weight were used, samples were drawn
at thirty-six and forty-eight hours. At thirty-six hours the blood
plasma concentration using 1 grain per pound was 3.7 mg. percent

while 7.7 mg. were obtained using 1% grains per pound of body weight.

-24-

At forty-eight hours the concentration had dropped to 1.4 mg.
percent with 1 grain per pound and 5.0 mg. percent with 1% grains

per pound of body weight.

Graph 1 presents the average sulfamerazine blood
plasma level curves following a single oral administration of-%,

l, and 1% grains per pound of body weight in tablet form. When
these curves obtained on canines were compared with curves on

human sulfamerazine blood plasma levels (Welch et al., 1944) ad-
ministered in single daily doses, they were found to be quite simil-
ar. Comparable results were also obtained by Welsh et al., (1946)
when they used-% grain of sulfamerazine per pound of body weight in
a single dose per 03. In mink slightly higher concentrations were
found using a single oral administration of-% grain of sulfamerazine
per pound of body weight (Langer et al., 1948), while in foxes the
concentrations were slightly lower (Welsh et al., 1948). The aver-
age plasma concentration of free sulfamerazine in cattle following
a single oral dose of approximately 1 grain per pound of body weight
was found to be lower than those found in the dog (Scheidy and Till-
son, 1947).

The data of Graphs 2 and 3 present results obtained
when similar doses of powdered sulfamerazine and an emulsion of
sulfamerazine were used.

Graph 4 shows the average blood plasma level curves
following the single intravenous administration of a 6 percent
aqueous solution of sodium sulfamerazine when-%, l, and -% grains
per pound of body weight of sulfamerazine were injected. Graph 5
presents the average blood plasma level curves when a 6 percent

aqueous solution of sodium sulfamerazine was injected subcutaneously

 

J

I
I
I
I
l

”I

—--——_ 1I- —-—-- n—oq. I‘l-

 

.-
< H . .
_ .3.
:H
I ..I- I III . Ii
._ w
u. . . T
-I II- _- -I...II4I..--II-IIII I --- I

 

_lI. I I till: IVQIII-Io o'.

 

 

a
_
_ 3
. , . . .
. . I _ . . ,
“H I... .4 :9.\. q ~43II4-HHQ 4.: .HH 3241.....31373:.II.H34.2441 \phumgm
I.- su (...-,1 .I..-. r...r3 u .23 -52»; .1. I. ~r (...... Spa-.3-.. LI. . r a .....nI-p x Cry-nu: I
o 4. a
.--- --. .1 -I a , .
_ _ .
_ >
.
<
.

3 I ’I|.IDI.III~ I -I I; .IIIII .II III. I All-II I II- I: I... .‘ I

%.
3

 

(1"! II 4|.ll-'i‘J

33.

& If‘

.. Lila-hr Mal”!- $2 kr««

 

I I_r I rt
« _ H
. .
. .
L . ._ _
. _ n
_ . . .
l _ .
I114 I'IIIII JIIII {.oI l_lllvc. I II II IIIIII T

, _ . _ . _

._

_ H l
w

aIIIIII-rIIIIIIIIIII III I III...

 
 

-| III- I'Il I11:

 

II IIIIHII -.II .11 IIIII.I|.I_|I.-III I! H- II III---.

         

 

 

 

 

 

!
1
I

T

' .
5

I 9-.-. ...--—-—_
I
- . v o a.

Imu—
l
o- _-.
. I -
.
, A
a

 

 

 

 

L L? .. ----v .-
4 -_
i
3,- kHz-xi . II---
M., . m” H
_ 3 . H i .-
.. - _ _ .
.o§£¢&3§¢Mr-IIII - - -- ---I
0.23.; tug Scat-O. _ _
. _

 

 

 

 

‘ 1mg

 

 

 

 

 

Pd“
J.

E
I.“
I

 

 

.27-

 

III---—-. w—I -—'q

"a?

find.)

“I? h: 4..
>1. 1»...

III III

Had-nan

 

L1. Jlufil' IOIHIYOIOI3 .Ifi.‘
...Iatk-HJOOWfi

_ .

.

.

«an I.....w ”-3.5; 433
“I.- ....In. ...: 50.3..

7...}...- ..fl

I. lo‘r

II . _. II * . . I —
.

«IIwu 5111.. $1.. afim. 4 n) 3.} 14..-3II3.34I1.. 711.. 3.043.:
.7.- .IIvs ... 15F . .7." -.n.» r... ..Ir..-3.....4I.I..-. 4... I323.-
_
. . . . .
_ .
o .

. --mmaari .

JV.Iw11II ‘
¢~aI

II. ICE;
13;: .-....«W.

.2 .4434?
c .HHC (:0. I.
.
3 u
I II. I I I II Ilrt

 

IIIIIIIIIIIOII I'll III:

M
_
.
...H
....L
w _
.
._
_

. . .

-

~ ._ . _

* a .

n . . . o

3 _ .

. _ —

'[llll'lll' I II 0‘ 'I I.‘ ll'tl’t‘ll II I II'IIIIIIII' III III 1IIIII I'll-III! IIIIIIiIlII IIAIIII II III III III In I
u . —
. .

_.

I
III

I

I

 

 

 

I
O
I

I“ *I v——- ~‘1 --.-—
I

I
I

d.--
.
5

IO- III

I
I
I
l
I
l
I

333 5.... 5.55 d III-IL
95... in: 23:9 _ TIIIII “

-253. «uh-ukir?w* rIIIIII -- - -

 

,3
p—-——_-
, . _

.—

. -.w
InII -..-II-“-
N _ U ._ .4
L . 23 ,-

 

 

'F’t‘. .

 

.28..

 

 

 

 

ii 4‘

 

 

 

 

 

 

 

 

 

 

 

 

 

~ _ . _ 4 . . . . u. _ _
_ _ _. _ _ u . 14111 4.41. ,
_ _ n n H n _ {Bum-.- C.
_. 1.- M? an...“ 4: Nam 1, flaw-Ham. «- ...H «film-48mm 1.2:: .HH... mafia-fl .- U nay-Xma- 41313 moo-H: «HUME-m-w-zwd-L-M” m. End-.14;- ._m ”mm-«AU
_ -.-.- 1. -- --1 w. L . H - - -. - -
n u u . .
. w . L . _ L _ h
-. p . H . _ _
1-11-. -- 11 ..--11- .111-.- .-11- - - -1 I.- 11- .1.- ” . --- --.-1-21.1- -1 - £45101? 1 ..... 41 - -1- - - - - 11- - _--- - 1111-
, . L fi #- _ am 2 2 z A. 2 u w w. g _
u L 1 - -.-. .- . L. .1 1-. 1-~ C . - , C #11 1 L L. M . N . - - 11
- _ H . q 1. J- . .. .. - _ . .
-- u m . u -- ..
___H ._ L . ., .
. _ _ . _ M ..
.-.- -- - 1.---1 41-1-1 - -1...111 .-..-..1.--. +2 .---1
_ u m _ U H..-
u _ w T"'-"‘- N
_ . . D
_ . ‘
1 1 . 1 -11 L -1 - . - . - . -- 1.
. . .- 0 . . N 1
. . N a . .1
q - . H a : .
.. - - 11 “11-- -. - - . 1-1-.1--1--.-1-r:---- - 1.1 R1- 81 -1-
”. _ _ ~ 8 _ .- V
v“? _ u _ . . H
L L _ _ _ L N ,
u -— _ a _ _ . . . . . 14-
.. . . 4 . s
:1. ., --w.- - . L L 1 .1 u - -- Kw}- - .- ”-4-
L. . _ _ _ . _ --_...- _.
m. u _ .- - L pd .
_. r11 1 o .
L . L ..T ... a
. L . u ...-4.1....- 7 .
_ m _ - m - .M
r11 ..- I 1 -_- 01 1 ol--- -1..-1 - 1- $1. .11 1-1110411]
. o . l
m H
. u ...
.11 1 - - - - - - --- -1--1 1- - -- - - $1 - 1..-- - L
n _ . _ _
_ .
_ . .
n _ _ . _ _
_ m H . . u I w—
L. 92.5: #2“ zECG-s 1:111...- . . 1- . - - - -. ._ - - -- - - - - .m.-- - - -. 1
m . . L . m h
m 013‘ run 27:» _ 1111...... u _ _ m . _ _ .
. _ .
. _ . _ _ . _ .
a-..b2393k933r¢£1lil11 - - - - - - i - - 1-- - _- ....... - -1 - -11 ----11-
. - . _ . u . a . .
u . a . - . u n m
n . . . . m h n _ _ .
_ - _ . . w _ _ n. _ _ _
...-.1.- .. - - ...: --1 .-.1 - - J --.11 - -111- .u - - . - - 1 - - {117- 11 1 H .1 -JI - T- 1-1 1-1- 1 . - - - .111.- 111-11 J ..... 1.1.-
.. . m ~. ~ _ . a _ . . . . u . n .
- _ L ._ . _ : L _ _ L. _ L m L: m
. . _ .L _ M ._ — - _ ‘ L h . _ L . ._.
. . a . . . u. _ . . ‘ O
11 .- _ Hfl— F _ .._ .2 L b -. ._._ I _ ~11. __ - - . H .1_ 1.9-

 

 

.29..

 

 

 

 

 

 

 

 

 

  

       

 

 

 

- - .
1 .. i1 I . 1 .1 1.111.! ...-
—.. . . I I , . I— . . H . 4 . ! . .11.! _ . I I]
I_. . I . .. , I _ . . . . I . _
_ . . I . I . I
u I . ~.
*.I..1I.n1 ...,Jflrh 141-11 .0 Inn 1.11... .....n. w .HH.).1!-...11 11.1.1. 31.14.. .4 x. 11.11.1111 1 . 1.. ......1
I.... 1.....- ....L -.. .11_..-.O. L1..1-H.I.1 .I. p C. 1- ALP-13H .mi. ...-”11...; .meEm I ...-1.151 41..an m GEM».- M-HMLH...dn...-J..1-UH,IW- ..I1.. . ......\_I -mmanms
l1. .h. .11 1|, 9‘ ' 1. VI I 'III Ll I — I l I . . ‘ -
I- _ _ I I a _ «mamI - -- 1. _ . - - -
I. .. .. . . I
_ . 9. 2. R a .I. ow .Iw on . w .
_ I .
lift-...III o t l- . . .I. .I .!|!III ’1!!! I I: ”V! . b . _ . . _ m m~ d~ & . § .
“I I- 11..- II1.. .I.. . ..4 .4 E P. . I O:
O . O ‘ 41“ I H 1.1 -l .l 'l .--!
.. I .. . .4 f'flt‘IL . . I
.u . 1 1" I
. - . . Olr . _
u . _ _ Ir _
_ I . . .
-1 1 - 11 -_- - . 1+ .
. _ . -11-- 11.2.... .-1 - -- - .1 -..-1.-
. I _ . I. I
1 I I .I _ I ..
. I . .r
.1 .- 7. m
. . a- I --1 - 11 1!...
u _ .0 fl _ .-
I _ . I . N
. . A -.
I m _ .d I .L
_ 1 ---11 .-1. 1. -1 1 .I. I - . .
._ I - . a. I I - - - 1?. .I- - -1111 1.
_ _ . _ .I . . II. .. . _ I. . 1 .-
. I I I .. w . . .I. I . . 1 7.
o w .- I m _ .I. _ fl . .' . 1. .
.u 1. u _ I . I. . a ..4
Y‘lrlc‘tlleI. 01!. .l I. -.llll v. Ill!.llll¢l 1... I! I F 1!!!! l .19! 0| 0.!) 1 ll .1. «II! I I III!!! I - ' . .9. .
. . . I _ _ . I 1.. ..l ....- -11l-1-1.11111 I111! .111! .-1. .1 II 1111 111 .1 1 30‘ .. . I
.u I a w I .“ . I I . . .. .VUI!!!
4 I _ .. I . u . . .«l.
. _ . .. I I . I . . .. .. . w
I... I .I. .I I _. I . I . 1 I .I x .. . .H
. . . _ . . . . . . o1
. I . _ _ . _ . .. . a . I, 4-111-71111-1. .
I- I I . . I I , /. . . ._ - ..
, _ . . . . . . I I I, . . . .I.
: .I I I I I I. I I p . I U _. . .-
fi1 _ 1“ _ I. I _ I . . /... _. I. I .a _ H1
1.1.1! 1 - 1 111.1 1 I!!! I1- - I 1..-“ .111 -1 1! .11.!1I-1+II1- II _.- I... IN! .- .1 1. 11r!l1, I- 111.1111.-1I1.1|11-.1M| 1.1.1! .11..- .I1 1W. - .-1! _ 1 a ._ .1.1
I . I _ I I. .I. .. I . . . - .I. --.-1.1-1111,-11.-II.%T-.1-.--1......
.. I I. I” . I n I /I I H ....-L
I u o _v . ._
._. .g .. I . .u c I w o .I - .N . ..HIAI
1. 1 1 1 1 .11 111.. 1 ”1111.11 1111 1 .11! l 1 1 11 1. 111 1 1.. ._.1 11. .1 111 1! .n 11.1- 11 1 1... 1 11.11-43 11. .1 1.1 1 1 1 I. 1 1 11. _/ II . l I”
. I I . _ I . . 1 . .I1 11111 1.1 . - .-.!!!11! ..I.
.. . . I . I I , .
. I 1 I n . .. . . I .
_ I . . I I . . . . I . ..
— 1 _ . . _ .u .I I _ . . _. -. W
111- -- - -I- .- - .111- 111 1.71 - 1- - --..1-1-11 1. -..-11 -1 I _ . I _,_/ I. :
_1 . _ . _ I . .- 1 -- 1-11--- 11 1 .I.--111 11 -.-1 ----1-1 .111
. w _ _ . _ _ _ . I _ / . .
I . . .
- - I _ - I I ' I \
- I . .
n u . _ N n . .( u .. . A
...-:- nun-N15 race-x. 11.-1.1.1. - 1 I- - 1 . - - 1 I _. 1. x I
— . . 11 11 .- 1 1. - 11 :1, 1 1 111 .-- 1’ ---: .ljf- -1.. 1 .1-11 1.1!...
- . _ . _ . 1.. _
u - n . l
a . . . I _ I I M I . .
I C o o.
.- - . nag-rut sf ....l...l... -. - - 1 - I . . .
. .H - . - . _ I . . I I u . I J
_ I m . 4 _ _ Q . _ . .- . — .— .vl
_ I I I I. . I . I . . I . . . ...
w , I I . _ I I .. I I . I _ _-H
.11.. 1 -11-1!1411.1-|1-.1-11 .. .11_.__-11..-.11 -..-1-11..-- I-.- . I I . I H . .I I . I . .1
. _ . . 111 .- 1 -l 1.1.! 1.1.- 1 - 1.111111 11 1.11 111.--I11 1 - -_ - . .I. . I L .I
u . W I , w . o .. I . W _ I I I . M . m . . m . w I . _ . I . . . . h... .1). v t I! J11. IA 01 ..AIAI ”I. . v! II. 1H... ll ! ir- ... . .llplolflfl 5.11mi] IoIWIOJIII’A
I I ...-.. . I . __ _. ._ _ _ I _ _ _. .. . I .I....
.I. - I ._. ..I.. “1 .I . w .I.. .. —.- _ . .... A o I . —. I . .. . . m _. I _ I .._ I I. 1. “In . I L“
I..L_. TLII-III . ... LII-.. .. .—_I _._w *. ._II I. ... .I. _. .. .w . I. .H. .I.-..III. . II.-._..... ._I. I ”Imm.
P. I. I. I . ... . .I..... In ._- - .«. . -. A... . .1. fl .. I .. “I ._.. ._ . . . . 1.. .I. I ..II . ._.-1
- . -. - 1. FPL.: . I. -II 2 I- 71.. . :27... I. I. 1; I .... .....: I... I _ ...I.1.H.....I..I.:1_IN__.1I~.
r LFL 1P» 5 b _ LP.

 

 

-30..

 

_

ALE-Em I

_
..mmmaewmb 1 war-I Mmon,4n-1rmm 4 1% 6T... __4.I.-mmHnMH.IIw-m mag (£5.36 rah-fin aim-MSW. mooggmbfimflflfl Pom.
‘. u _ - ‘ a . I .

_
1|! I 4 I4 I II :III II I II . - I vIII III I . I

.. . _I; l ._ .m. a . -A _ LII. I. . 4. I1
. . . _ _
..

a1
J
:i
.3:

I'
I"

a

    

.

I
I
l
I
I
I
I
I
I
I
I
I
I
I
I
- - 1---
I
I
I
l
I
I
I
I
I

 

 

 

 

 

 

 

      

 

I
I
I
I
I
I
I
I
I
I
[I . .
I
I
I
I

 

 

 

 

 

 

 

” _ fi ll. .
WIL- I I I I “I I . II. I- III I .. -I I II -III II --.I . -. - I .L .I. In . .I’ I . . .
THI . . _ . . _ . . I_ n . .I’IO . _ .
I-r _ _ . ~ M _ . _ . _. I If. _
_ .
... . n . _ — H -_ _ fi _ o. u /-I'
II n . . ~ _ n _ _ . . _ Ll
_ . _ _ _ q . H.
WIIIIIIIII IcIIIIII IIIOIIIIIIIIII IlmIIullIIIII . III-II III-rIIIII _IIIIIIIIIIL III) I_ IIII’IIIII.|IIIHVIOIOIIIIII II
T I _ . ~ _ .
.. . _ J _ . H n . _ _ H . . I .
c . . .. _ ._ . I
rl .* . .a. . w . _ . . . ‘ . .. .
1 . . _ ~ . _ _ . _ .Iw
‘ . m _ m .__ . _ u _ a .— _
If». I l _ I II_I I it. I.vllhII_I Q. .I «I! IIIIIIIII-Ifi IIIIII- .I.. ..I II I I 4| .- I .II I I II IIIIII-I III..- 1 I T
f... . a.“ _ _ . — a _ _ . _v
- . .. I .I w u w . . . .
. w
_w .. I * I _ . _ u . _ I . _ _ . . . ._. . ~ I I.
_ . . _ . . _ _ . _ . _—
m .. M f I _. I fi . . fl .. .u m. _ _
_ g _ _... ...... _ + g . I .-n . _ H . _. -
. ‘ _
I+II1IIIII IIII II I. I.Il_oIII|bIII II I I I41 I IIIII. IIIIIIIIIIII III IIVIIIIIIIIO III] II II IIIICIIQIIIIIII |IA III‘IIII 1.! III IIIIIIIIIIIIIII' IIIAI IIIII IIIIIII IIIIIIIWJII l
I I l.—\ . _ m. . . _ _ I— a . fl ” .
x... , _ I -- _- u 2 fl “ ._. _ . _n . .I _
. — h | . . .
a . - ., _ . w . . w _ . . _ . _ _ I. V. _
I . _ .0 . u . _ . I. _ . . . _ . _. I .I _ . u I.
. _ _ . _ _ _ a
. . . .. . ‘ _ n c .I I. . . , .
. _ . . . — t. .
IOOI l I I VIQII «III IIIII Jill I II IIIII II .II I III III-I .I IqIIIIO II IvII IIIIIIIIV I.IIIIII—.I| I II] Io! I II III [Iv-IIIIITIII vII OILI I II IOIIIII. I I I III
_ . . . 1 . I u . I . _ m _ .
. . n I M j . I . H . _ . g . _ n ._
I.I.. _ I. n .. N n I _ . n a _ .m
. . > w w _. _ . . m .N H. I. .— .
I III I. -II -I I- I- I- I ...II--III III -I. . II - I-III I.” .I.! II-IIII. II-I IIIII -.. _IIIII.-.I- III.II. I. II I- III III I _III. I-..
jI . _ a ._ u n 1 I. .. _
_ . _ _ u. M _
n I _. _ . _ m. _ _ .. _
_ . . .- . . H . _ _ . _ _
.- . ,_ . . I. . _ . _ . . . q . . .
8389 emu-g II-: -. -II -. - -- -.- .- _. I. I --- I I - ---I -I I -I -- III I .
. . _ . . _ d .. _
. . , _ . . m n
. . m _ u _ — m . _ 9 v _ _ II
. . . . . . m m a .. . _ . _ _. . .4
_ . .. _ . . H 1...?
.352»,- In». 52.5“; .I I I I - --I- I: .I.- - -I I. I- -L I -I- --I - Ti. I. I I: - _-:I-I-:-_..I-4 ----II
. . I I . . . . . . .
”.... . . I . _ . . . ._ m . . m _ . .. _ . . _ h
_... . _ . . .. n . . u _ . . H . ._ m _ _ . n g . . _
~ _ _ .. . _ . I . to .a
_. . n . . . IT . . I . __ . . _ m“ 1 \ _ _ _ _. _..
_j- I.I-.-.--It "..-.IIIII;I-._I..II-I.I-,r-I.I-II-.-.-_ III-IIEIqu -I.-.I. IL- III-III ...-.I- “.I.-«III? ,I f .I.-IIITI -I -.-r .I I III . T II_rII II ._.1 gm"
.- . _- — . .IH a . _ a . a . — . _~ . ‘ ~ III-III .IO‘I‘I‘I— . III .—I; _.
u._ _ I~ ...—- _, . .. ._ .. I -_ ._ .. n . _ ._ _.. _.,_ ,. _ _ _.. _ . ._. my
..I. . . . . ..n . q . — . .I .. ”- ._. . . .. H _ . I . _ . _ . “ “ LI. _e
. .-.. ... . .I . . _ .:_ _... _ .. __ I. _ .. I .. _ _ _ . _ . _.. .. .1.
.. _. .....t... . . _ . .._. I... H . . M _. I. ; _ M I. ....._ I“.___:...u
H.” .j .. _H- ._. .H. ... q . . ....‘ , _. I. — ,.v. H »-o a . ._.__ 9— _ ."I . o.—_ .o . __ . , I” ... l. > _—I _IHI%>QO

-31-

in dosages of i, l, and 1% grains per pound of body weight. When
the sulfamerazine blood plasma curve on canines was compared with
the curve obtained with cattle (Scheidy and Tillson, 1947) follow-
ing the single oral administration of-é grain per pound of body
weight, they were found to be considerably lower than those found
in cattle.

From a clinical standpoint the primary interest was
1) to determine the route which was the most convenient for adminis-
tration, 2) to determine the route or routes which were equally
effective or advantageous in maintaining therapeutic sulfamerazine
blood plasma levels, and 3) the frequency of administration necess-
ary to maintain therapeutic plasma levels with sulfamerazine.

After analyzing the results of the three routes of
administration, the oral route seemed to more nearly meet the clin-
ical qualifications required. The intravenous route was temporarily
discarded because of occasional difficulty in administration (object-
ion by dog) but primarily because of the large volume of 6 percent
sulfamerazine solution that must be given to maintain sustained blood
plasma levels. This latter objection is also encountered in the sub-

cutaneous use of the 6 percent solution.
SULFAIERAZINE ADKINISTERRD SEVERAL TIMES PER DAY

The next series of experiments were designed to deter-
mine the sulfamerazine blood plasma levels when the drug was admin-
istered by mouth once daily, twice daily, and three times daily for a
three day period using %, l, and 1% grains per pound of body weight,

respectively. Studies on the blood picture were continued.

In the series 01 eiperiments where the drug was ad-
ministered once daily for tIree days the following dogs were used:
5 grain per pound of body weight - 106
1 grain per pound of body weight - 109
1; grain per pound of body weight - 112

Tables V1 IIVI, IIJII, and IIVIII.
[hen administer ed twice dai
ing dog 3 were us
~L grain per pound of body
1 r'rain per nouns of body
1} grain per pound of body
Tables VII, I.II, III, and (III.

ly for three days the follow-

107

weight -

weight - 110
° «1 ‘3

11919.11? - ll.)

When ad minis to red tEiree times aily for three days the
following dogs were used:

§~ grain per pound of body weight - 103

l gi‘a; n per pound of body weight - 111

1; grain per pound of body we i‘ht - 114
Tables XIII, IC'ZI" , ICZIIII, and IIZIIV.

Graph 6 shows the average sulfamerazine blood pl sma
level CL ves for the three dom es wlien “ruinistered once cm ily for
three days. Graph 7 shows he for twi e dai ror thr days
and Graph 8 shows the blood pl sna level curves wlen sulfanerasiae

f"
5.3V

‘Ias a.1—. ti..-“

red three times daily for three days

”he sulfa:ere.zine blood p_asna level curves were found
to be quite regular. The height and maintenance of the smlfanerasine
concentrations were also found to be directly proportional to the
dosage alministered. When sulfamerazine was administered oralLy once

-33-
daily for three days it was observed that a slight increase in the

blood plasma level occirred on each succeeding administration since

normally ahout 80 percent of the filtered compound is resorbed by

the renal tubules of the normal Log (Peters et al., 1944).

.msnp 039 me soapprmwsflE©s someone mophuapsfi *

 

 

 

 

 

 

 

m.e 0.0 m.m we
o.m m.e m.m co
e.m m.> e.e on
e.© H.m m.e mm
e.m e.e e.~ * we
m.> ~.m s.m om
o.m w.p m.e mm
H.m e.o m.e mm
m.m m.m o.m * em
0.5 m.m m.m NH
o.w w.o m.m w
¢.> m.c o.¢ e
o.m e.m e.o H
m .mg m .ms m .ms
as a .m
schnappmaewaee
eosmpmaeaeee season seen .pfl\.no. “meme muses .

 

.wwacosae as NHH .ooa .eoa .02 neon .maen mouse wom.sflwaa ooeo
enfinwpessmasm mo soapwnpmflsHEpw Heao emu MQHSOHHOh mHo>eq namwam nooam .H> eHan

 

.3 5..

.msse one no cowpwnpmwewaee nmspoas «assesses *

 

 

 

 

 

 

eonopmagwsea pamwoz_seop .pfi\.nc

 

m.©m 0.0N m.NH Nb
m.mm H.Nm NowH ow
m.mm N.bm o.mH * om
o.©m m.>m N.mH mm
w.¢m o.mN m.ma * we
b.0m m.bN ¢.©H om
m.@N m.MN %.HH * NM
¢.¢N ¢.©H momH mm
m.bH w.NH O.m * flN
H.5H m.NH mofla NH
®.OH N.b N.m * m
m.m m.¢ m.¢ q
m.m o.m 0.H H
a .ma ‘ a.ws a .mg
“a H I.“
cowpwnpmfleasee

amend mnsom

 

.xweemama aw mad .oHH .eoa .oz amen

.mhdn mange

woe swede mange manganeseeasm mo gowpwspmwewsee Hugo map newscaaos mHopma «swede eoodm .HH> wanes

 

.36-

.msse map mo nowpespmwcfiaew seaweed «assesses *

 

 

 

 

 

 

m.oe H.mm m.eH ms
N.N¢ H.5m 0.0H 00
H.oe m.em o.oa * om
©.wm ®.Hm moMH # Nm
m.om 0.0m o.oH * we
m.mm o.mm «.ma on
o.mm m.s~ o.HH * mm
o.em N.Hm «.5 * mm
e.mm H.ea H.o * em
m.sa H.ea e.HH NH
m.oa m.o m.o * m
.Now COM COM * IV
e.m m.m w.H a
m .ms m .m& w .wa
“ma a m.
:ofipwhpmflsfisd¢
eonopmweaeee pawns: seen .pa\.go amped mnsom

 

 

.xweamsae aw «Ha .HHH .moa .02 amen .maen amass hoe asses
mmsfle mouse egaushmssmasm mo GOprnpmfisflEd¢ Hmno amp msHSOHHom mde>oq msmedm uooam .HHH> manna

 

.37.

 

    

 

. ._, _1 in _w . . 4 , . . . M . . n I.
_
. I .. .. . _ _ _ . . _ . -4 _. 7444. H - I... 4 U. .1! J. . ..- 1. ....
.... . ., . k . M M . I _ whi!» r1??? ....0 Livafiermr 1.1.. '..c»-_ If... arts. .1... M rrchmknr VWFMM. debug...“ r74...
u, _ e u . _ 1 .a M
_ ... .. 34.14.... .... .. ... 4.3.1.. I). 1!). ) 31+ 3...... q :4 3.1:: . I ) 3.4. 4......144 in _ 1.. . W3: .1 ..SA
fl. KW —. m _rwwbrwrn . MSIRFL , w. “0...”... II! fPPM o: ... .- wH/k Mr“... Huff.» .ML...rmrfi. rrr... firmwaldol . 7‘ ”flak“... rlru 144.....9fiy ... «up. ...fl Paw /.3., .Hmmt \» MMWH l4?l-“n...1~vk...rf Pfi.
r . I A . . . ., . F. , ._ll _ . . .. -_, . . p . ., i . .... .. I ..ILOJI
. . . _ _ .
._ . _ . . M .. .7 _ . _ 4.
i . M M .u I _ M M . _ H
o . . _ . . . _
. _.. ._ . . _ . . _
n M. . .. _ . m . _ _ _ M I _ _
1. .3 'III '1! ..llfdlllu'lllol‘HOlell. lTw‘.’ u I .60' III). IIMIOI II II .Xll i I «I .ql' ._ I ..OOI. l.u I‘ll ll.] III... to: Wuil Juli. I -‘l '1'. . 'I I on I. .I- mil! III| _ . . 1UP]
i _ I , u. .. . .331» t. _ . _ _ . _
_ _ i.. M . . .. ._ .u I . . . . M _ .
.31. .3 3 «My . an mm .2. as. . .8. v, I _
r: . _. tr“: "3.1 Map.“ .... ._. . a. . . p I .. . . _. . _ $
d- 4 4.. ,4 — 11144 . _
-_ , _ I _ .fi. .H. a. I. M _ .4 . _ _ . . . -
. m . — . . _. . _
u I M m . I _
I ,- _ ._ i _ I
. n . . . _. I .
dust..- I- ..-n- .. ,a. .I.. 1 III! .Wllllulr- I'll. -I - -- 11.4.51! -v .1: -I. ---.a. - Ill..- II I- I...
_ I I _ I . . _
_ M 4. u M . .—
M M .- . I _ . . _
. H , _ . . .
1r. - ”-1 11' I: I f» l v. -.. ..-I.-JL}.I -I.; u .1 I l .. -
._ I . . _ ._ w .
. . I. .I a _
- .1: it'll?! I M . ; ill]!!!

 

u ...,q— _.. _—
- ¢

I
V
' 9
1

-_.-u-
...

._n:99$ zua.uad¢mvwfiiil!l

   

 

932.. «it. L.4¢&_ .. _.

l.-|' OI I‘LII'II .IOIII IA, IIIIII:
.

 

----—‘.-— _w J———-—.- _..—_..
u
~-
c

 

u
. —.—..—_'—..—-.._-—--~.--.

_: _r
_ . 4x
+ I a“
.I ...I
I l,— - [-411 I I.
i m .5
w. I.

 

 

l
I

+- -—-.1.-—— 4

. 'lll‘ loll- l¢

-.. -._..-- _..---

 

_ H
«n. In .-. ....n 10. .I- 0 c I n I. III _.III 110 6 .I- I. 10 «I 0|-.. 04] .I...- :19 II .| I ..lnl. 1|- 0 I. . IQ» . I. II -0 0 Alli
. _ . i . . .
_ . _ _ . __ .. I . _ _ .. . ”I.
_ . ail. _ _ _ m _ _ M . . i H-
— _ c u “a
. . — .
_ M I+ . .U a .n. ._ .« _ M _ M _ _.. I.-.”m
. . . . u . . _
_ . ._. . _ _ i . __ . _ . . _ . . _ .. . . I- I
L—M ‘0" ‘if Ill In“ 'tt'nt.’ .lol ,I 1‘1». Y} i“lolt|l* r-1li|.l o'ilv «it OI\I| .."IIll 1v. ' llaJl' II. .I. '1” vl II VI! tf‘ll lib . .I III vIOI' 41".‘0 I ‘1 .Olilolll. [Ir ' “Q‘OI tl I... lll.’,l4*.vlfll+l
v. . . 1 _ . i . . . v . . ..w
I . i .,_ _ _ H . _ . . _ .. . .. _. . .
_ M M _ _ a
. . M. .. _ . . _ M. _ . . q. . . . .... .9...
. . i ..M. i M . . . .. . .M . . . .... _ .
s .— v —. ,. . . .. _. ,o . . . . .. _ u . . . n“ . _.
afi. . _ . . _ 4 M. _ ..... . _ _ _ . ._ _ . . __ . , .. M . . . . . . a r . .4 .a ._ . at... .~
. M ...... . . a . .. . v _ .. . . . .. _ .. . .. . .. V _ . . i. _o . . . . ... _ . p uL.
. 4M v k y. . .M . t—~ .._.. M. ... ._. a .. I". . I .M. .. —__ .. _ _».. M ..p....._ ,..., ...—. ._ .~.¢ ... . _.. ... ,Ia.-w..“%
r r+ rer..s-i rrr. .» ...vre r... e . irr. 1.- , . var. hr. , ;. .r .-i- _ .. >+.-.M... , , . . . . .._ ...... .. - ..wisu.a

 

 

   

 

I
M
I

     
 

 

 

 

 

 

 

 

 

 

-ILI._

 

III _ .I ._ ._. .II fl I.I- .

w
. . . .. . . . II I I (I \II I r I! . . l-
. . I, I. _ . ......Im mama. _m _..... wast- ..-..WI. 2.8 0......IID I _. £83 .3 83m...

. MIA” HWHHH w. «HIlrd ELIE. III... LHMHIW... “.I-.I.I..._... upwwmh 3 ......m14 (WED «Beam .-IIMHMI..B nape... m..wfim~w¢ .N. mm.“ on

— ._-—...

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

 

“
I.I I II -.. -IquI..M II MI I I.l.l..H .. . , . I, . I I . IIII I. I l-l.l.I.,I I .. III“: I l. . I. II
.I I . .u I I I M . ~ I H _ I
_- _ I . I. _ I . I I _ _ I _
II... ......II. I. . . III . - I III III -I- ....-.i- .I- I “1.: I I --II II-.- I-.. ..
-H M _. I . .. _ IMI . .I I I I
_ . .. .. . . . I
I . . .. .. . I .. _. .. . I . I
3 . 3. .3. IE .3 _ , 2... fl 5 . 3
fl M . MI, .MH . I» .. ,HM .I I . . L MllLII I LL .I I _ w- I“ .
. - .I III... .. I...,I ....I. III ..IIII _. l- I _ I. _ I .I.. .IIH. .III.II..I..J.,JI - . ALI .I.--. ..I.
-TIIII M I WI... .-WII-..,UI:. -- .. Ind“ .I ...... Fm . 5: .....AI _. . I? . III: .HIIt WWI. -...IIMI..L ._.I-......
...m n; -mu.IMrIm,._I I ., I? ..-. MI . I.l I ..I ”I.I.II.II.H--1.I_ HI; . .I-.» .2...
I. I -- _> I... _wlfilI I III . ..-II .-..III II..... 9.1. I III I. ..-... ”I.-. ..I.II....I-HI IIIIIII
I. I. - rIII. , 9.393%... ....IIII l...l..II_ fluxI ...IH. IMII. I. H. I3. I. .II-. I n. . MWM ...IIII
. .l ... I. .l. .-rmML... I MW IL .I.- W IIH .t .I .I.-.- _.I III I I .I.- I .I.... -.-- -_.
4.. IHLHI .. I _ _ M I M _ M
-. , - I _ I _ . I
_ ._ I
. I I M . I . . I M +19 I
. . I I . .I. _ I I . . . \st ......Ir
- .. . I ._ I _I ..I..- I - I... I -I-. .. 2M -..... .-....meit --
I I . M ._ .M . . .. . M I .. x l'.‘ \
I M. I . a . M . M .. ‘ ILIII I
Ink . ._. _ I.. _. I. , .. . . \.. . . ..IHnu. .
II. M I I .o‘fli. . . . \. _ . IITI .
IIIV I I IIII I I IIIII .Illf'j.‘ “I“.I‘W. IJI‘II’IV‘II'. :I'II_. I I III: .II‘ MIA M I I._VM .I
. I

 

III A I.I|IIII II III- III

  

 

 

 

 

 

. .MM-MMM_ _—._. M- _-

 

 

 

.
.
_ I
I
o
. I M .
I I I _
. - . . . . .
_ _ _I . . o
I .I..II II.- I .I .IIII . III... ...-II . . I..I.. ..l. . I I II I “I. I II III ..II II I I .. l I I III .I.- l! I I II J- . I I I I L.I.oI III IIIII
.

 

 

I ,.
I _ M
M I M M I . . _ M I . . I I . . _
. . . o
. M . . . _ I M
n . M . . — M M I M I . M M _
I . . . M
. I I. . I . I _ I I _ I I , .
~II4I II .I I _I. I I I00 I. IrI .I. I I. In I. III II *lu.‘ 9 I .vl II. II II IIII. I .lv.Il. I¢I .l M. I... I II I— I I IvJIII. cl II..II| III. M II ..III . II. II III I . III I I 1 III JII‘IIIIOIIII w III II. I“ . IIIIII _ . _I _ .-
— I I M . . M o M I _ 1.....IIIIIIIIIII .I III ..IIIIJIIII
.. v . . a .
‘ _.. M M _ M . M . M . I .. .M I II M . I. . _M . .I M .\ M .I
. . u . I . I . . .
_.I. . .. . . _I . _.. M I . I _ . I I . . . . . .I ...
Mi . I _ . .I .I. ..._ _ . I _ . I . I II I I. _ .. I I .I .I..
9.? .. . o I . . . .. — .< . M I .I _ . V M I u .I. o . .
-.__. I _ . ..I. . _M . II. _..... .... .. . . .. I I I I . ... I..I. .I .I.

 

 

 

‘1IIII'I ‘

I . I - .
_ I
”4“.)4444 {)I-IC I‘ll-4404.4 “(VII .44. .---.44.! 0.1
M i .I. ...u -I... 1 r; , _TtI... .....ZIC n..T.OI.....Q D u T....).....
M. . 441.4% \4440WoMONI .IIIL I44 4...”... .I.. a...l.1\1.. 4.. 11% .
4.44.. ..k.\l.)IH1_ 0J9. mf\.r4 Hr,

.r._
I

‘44... 4.044.- III 4

.71
»———I
.4
—-1

M .IOWI)I.4M.II «Hu44IrIM.Mv.
c.

«I... .14.: ...-..

., 5;. 3313.1... ...I........I...I..
IIr--IIr rrtvfll _( ..r ...r7M

“If“ (II-OrDL. "4rrhrr’hy'tofihrluvfl {3! l\. IDIK
—

M
M

I . M . . . _
_

II It... II|IIII.I In IOII. I I4

I
A
I
4

[4

148.4,» ..WIIr n hr ...._
.... ”WV/Cw. .4 ”(H-
l\ v:

.u Ct

 

 

 

""7“?” --" “ "—1
~92
3
:3:—
IIIIg I .
if ' Pg 5 .1
‘3
-—-:
g.--
«7:
E
E
[E
3-

 

 

 

 

 

- . .-l.-..r\mNI-I
._ . \
I .. ' _ . -
I \ I .I
M
. I
II\--I.II|I.1 ---..II I- .I. .LI I II ...IIIII l.
I I
. . I
, I .
. _
I .. .I

it

0534c£.%36M _

 

. I . . _ .

- .
. .
. M I . . . I _ I

‘ V
..MM- M... _- -
‘

_m' -
.I M -
I
..
M
M... ....M

IW .92331 (at .gfiwli .

 

 

 

WHNVH9?

lNEQUEd

i

- 1

ITWIhJ

 

q

10st concentration found on the second day of administration

0 1 v. ‘ 3 O
var1edsl1 htl;r depending on the dose e USOU. Ultn‘g grain per
po.nd of body'wei"h , the highest concen r1tion was observed four

hours following the second daily dosage when 4.5 n". sorcent was

LJ «-

f 1* .4 *T°"“v1 7 "1" i 10‘“ 'm '11 1 ‘ 1 '7 fljn “us-11.37 1" ’ »’ 1n” w -

01nd. wish 1 U1ain 3-; -o1nd, a 01001 plea- -1 _ove- 01 0.1 LU. lo,-
I J— J-‘-: ”‘1 . ‘: d 'N - ' '

cent was obtained at n.1s sane ‘103413 111 .1 s 1111.1 level has

n1intaincd for another four hOLrs before a 'ecrease in concsntration

'1

111 sna level continued to

.L ‘

was found while rr1ta l " ains eer p01nd she

-. w J--. : 'l‘ ‘| I" D .. 1o , m - I... 1, —. -» ,17.~-:~ . "q " r 4"! .
rise anvil e1bnt nour1 1ollow1no the second daily aa11a11trie1on wien

Fri

a concentration of 9.0 ng. percent was found. ollowing the third

W o '1 o o

a 11y a iinistration, a concentrationo f4. 9 m”. percent was founc in

p.

Q o J.” 1 o ‘ ft. ‘__ .0 -- _‘ ~ .... ’5 v
four hours Ulbfl'J grain saliaueraslne per s01n1 of ood' wei: :1t a1ter

which time the plasma level adually dropped until 2.5 mg. percent

was present MOI n -+ JIO honcs a1ter the initial dose had bee: given.
With 1 grain per pound of body weight a concentration of 8.1 mg. was
obtained four hours aiter the third daily do use e hcld sec n adv; :istered.

This plasma leve. eropeed slowly so that 6.0 mg. percent was still pres-

ent: eventgr-two hm ”s follow'i n; the initial dose. I*sing 15 grains per
pound of body weight a level of 9.4 mg. percent was obtained four hours
after the third daily dos This level was naintainel fo: another

four hours after wh h time there was 1 gr adixl dr0p with a blood plasma

level of 6.3 13. percent still present seventy- two h0113 aiter the in-
itial doe). These data show that to a preach a cone 1-tr1tio n o; 5.0 m5.

(4

, ”\‘L 1 v- {'0 ax ‘ 1 n.‘ ‘ " "1 1 ‘ ‘L .“"' .f‘ ‘. ‘ ,‘1 ‘ 1 V
percent, anL fire1eraal; aoove o nu. Her Sly twena-—1011 hears a 11],

1‘ o _ o o, ‘1, +V p. ‘ ‘ ‘ o '_- M ~
w11c1 1s consilerea the enerageitic blooa {l 1sna lerel (Jels h et al.,
1946 la "“a’r" o? snlpnterosire oer poxnd of bod;r w11~ht woe 1000"”—
._., L; O"- .. -D ..h -L...J. g... -v‘ L‘s—J... Av ‘ —/ ‘V—L U . ..H a ‘1 ads)

ary when administered in a single daily dose per os.

.41..

When sulfamerazine was administered by mouth twice
daily (9 A.N. and 5 P.M.) for three days the following results were
found: The blood plasma concentration made steady increases with
both 1 and 1% grains per pound but-% grain per pound gave an irregu—
lar although constantly increasing plasma curve. As one would eXpect,
the highest concentration obtained was found at sixty hours or four
hours after the last dosage had been administered. With-i grain per
pound of body weight a concentration of 4.5 mg. percent was found in
four hours while concentrations of 4.9 and 8.9 mg. percent were found
with l and 1% grains per pound of body weight respectively. Eight
hours after administration the blood concentration with-% grain per
pound had dropped slightly to 3.2 mg. percent while the concentrations
with l and 1% grains per pound was 7.2 and 10.9 mg. percent. At this
point a second dose was administered causing another increase in blood
plasma levels so that in another four hours the concentrations for-%,
1, and 1% grains per pound of body weight were 11.8, 12.8, and 17.1
mg. percent, respectively. Twelve hours elapsed before another bleed-
ing was made. At this time the blood concentrations were 8.0, 12.8,
and 17.9 mg. percent using-%, 1, and 15 grains per pound. Another
dose of sulfamerazine was given immediately. Four hours later (at
28 hours) the blood concentration had reached 12.5, 16.4, and 24.4
mg. percent. At thirty-two hours blood plasma levels 11.4, 23.5,
and 26.5 were obtained. The second dosage for that twenty-four
hour period was administered. At thirty-six hours blood concentra-
tions of 16.4, 27.8, and 36.7 mg. percent were found; and at forty-
eight hours blood plasma levels of 13.8, 25.0, and 34.8 mg. percent

were observed. Another dose of sulfamerazine was administered at

.42...

this point. Again a slight increase in blood plasma levels were
noticed. With-g grain per pound the plasma level was 15.2 m3.,with
1 grain per pound 27.9 mg., while with 1% grains per pound 36.0 mg.
percent was obtained. In another four hours (at 56 hours) the con-
centrations were 15.0, 27.2, and 35.3 mg. percent respectively. The
second dose for that day was given each dog immediately after bleed-
ing. At sixty hours blood concentrations of 18.2, 32.1, and 39.8 mg.
percent were found. This represented the highest concentration
obtainable with two administrations of sulfamerazine daily. The
following morning, at seventy-two hours, the blood plasma levels had
dropped to 12.5, 29.9, and 36.5 mg. percent. The data reported herein
show that therapeutic blood plasma level curves may be obtained with
the administration of'% grain of sulfamerazine per pound of body weight
twice daily to normal dogs.

When sulfamerazine was administered by mouth three
times a day (9 A.I~:., 1 10.32., and 5 RM.) for three days, that is, 11:,
3, and 4% total grains per pound of body weight daily, the following

results were found: With the exception of the administration of-;

grain of sulfamerazine per pound of body weight, there was a very

)4

steady and increasing blood plasma concentration. When-r grain per

(x.

pound was administered there was an irregular although increasing
plasma level curve. Again it was observed that the highest blood
plasma concentration was found at sixty hours after the initial dose

had been given. Four hours after administration concentrations of

3.0, 5.0, and 8.4 mg. percent were found when sulfamerazine had been
given in dosages of 5, l, and 15 grains per pound respectively. Sulfa—

merazine was again administered in the same dosages. In another four

-43..

hours blood plasma levels of 6.3, 6.8, and 10.5 mg. percent were found.
Ag ain sxlfa merazine was administered. At twelve hours blood plasma
levels of 11.4, 14.1, and 17.3 mg. percent respectively were observed.
No more sulfamerazine was administered until the next morning. Blood
samples prior to another dosage of sulfamerazine gave blood plasma
levels of 6.1, 14.1, and 22.4 mg. percent. As shown by these figures,
the blood concentration when-L grain of sulfamerazine per pound of body
weight was ad ‘.inistered was eliminated rather rapidly. With 1 grain

per pound the concentration remained the same but when 1L ‘rains per

4.

pound wen“ given, the blood plasma lev vel continued to in crease. These
results are comparable to those ootained when su11.rnnerazine was ad-

minister ed twice daily in the same dosag es. The first dose of tne

second da3 was given. In four hours or twenty—eight hours af er the
initial dose had been given the blood plasma concentrations respect-
ively were 12.5, 21.2, and 24.0 mg. percent. Another dose of sulfa-
meraz ine was administered rally. At 32 hours, blood plasma levels
were 11.0, 27.:, and 32 .0 Ig. percent whex L, 1, and 1L grains per
pound respectively were administered. Again each dog was redosed.
At 36 hours blood plasma levels of 12.4, 32.0, and 38.5 mg. percent
were found. No more sulfamerazine was administered and no blood
samples were drawn until the next morning when the blood contained
10.0, 29.0, and 36.2 mg. percent of sulfamerazine. This represented
a slight drop in the blood concentration of the drug overnight. The
dogs were redosed immediately after this fort3r-eight hour bleeding

At 52 hours the blood plasma level had again started to rise. At
this time the blood concentrations obtained were 16.0, 34.8, and 38.6

mg. percent respectively. The dogs were again redosed after the

.../.4-

drawing of the blood sampl 3. At 56 hours with the blood plasma
levels still rising, 16.0, 34.8, and 40.1 mg. percent were observed.
Following this bleeding the third dose for the day was administered
and in four hours, or sixty hours from the initial dosage, the high-
est concentrations were obtained. These blood plasma concentrations
were 19.0, 37.1, and 42.2 mg. percent respectively. With no more
mlfamerazine administered, the blood plasn concentration slowly
dropped so that at seventy—two hours after the ini ial dosage had
been give n, plasztla concentrations of 14.8, 32.1, and 40. 8 mg. percent
were found. Considering 5 mg. percent as a therapeutic level, one
can readily see that it is unnecessary to administer sulfamerazine
three tiIies a day to maintain therapeutic blood concentrations.
The same brand of sulfamerazine was used throughout
the course of this study, ir cluding the tablet form, powder form,
mulsion, and sterile powder for the preparation of intravenous and
subcutaneous solutions. All three types of sulfamerazine products
given orally gave similar blood plea nla level curves. The first study,
comparing the single oral administration of L, l, and 1Lg A rains per
pound, was begun 7:-arch 1,1948 and was completed Algust 6, 19/ +8. The
study comparing the various dosages given either intravenously or sub-
cutaneously as well as the oral administration of the drug for a three
day period was begun Aug ast 9, 1948 and completed September 3, 1948.
Blood studies were made at spec 13d intervals wi th
each dosage of sulfamerazine administered. The results of each trial
were further tabulated to find out if there was a trend towards lower-

ing or raising of the cellular constituents following the administra-

tion of sulfamerazine. It was found that no changes occurred in the

~45—

red and white cell counts, hemoglobin content, and differential leuko-
cyte counts when sulfamerazine was administered at the rate of-L, l,
or 1L grains per pound of body weight once daily for three days. Blood
changes did not occur even when sulfamerazine was administered by mouth
at the rate of IL grains (214.5 mg. per kilogram) per pound of body
weight daily in three divided doses for a three day period. Similar
results were obtained when the drug was administered at the rate of 2
grains (286 mg. per kilogram) per pound of body weight daily in two
divided doses for a three day period. Fifty-two hours after the in—
itial dose of sulfamerazine had been administered, when given at the
rate of 3 grains (429.0 mg. per kilogram) per pound of body weight
daily in three divided doses, a leuk0penia developed. The leukocytic
count drOpped 39 percent at seventy-two hours. At the same time no
changes in the hemoglobin content were found but a slight monocytosis
developed as the drug was continued. When 3 grains per pound of
body weight were administered daily in two divided doses for three
days a slight leukocytosis develOped in twenty-four hours which grad-
ually decreased. A slight hemoglobinemia and neutropenia were found
at seventy—two hours. Upon increasing the dosage to 4L grains (643.5
mg. per kilogram) per pound of body weight daily in three divided
doses, it was found that a marked leukopenia developed eight hours
after the initial dose had been given. This drop in the white cell
count continued throughout the course of administration of the drug
until, at seventy-two hours, the leukocyte count had decreased 60 per-
cent from normal. Hemoglobin changes that occurred were significant.

Although the red cell count remained fairly constant, the hemoglobin

content decreased markedly as the dosages were continued until it had

~46—

drOpped 28 percent, (Table XIIIV). Analysis of the cellular constit-
uents showed a merged neutropenia and lymphocytosis. With this dos-
age normal healthy dogs showed anorexia which lasted several days after
the course of administration of the drug.

It was interesting to observe the blood changes when
sulfamerazine was administered either intravenously or subcutaneously.
Both routes of injections produced a leukocytosis even when-L grain
per pound of body weight was administered in a single injection. No
major changes were found in the hemoglobin content or in the cellular
constituents except a very slight hemoglobinemia of a,froximately 20
percent when sulfamerazine was injected intravenously in single 1 and
IL grain per pound doses. Also a slight neutropenia was found when
l: grains per pound of the drug was injected intravenously.

Two hundred fortybsix sulfamerazine trials were run
during the course of this experiment. In 85 of these, blood samples
were taken at specific intervals on each individual trial and check
for red and white cell counts and hemoglobin content. Differential
leukocyte counts were made on 69 of the dogs.

The greatest difference between the initial and final
count in any one trial was 990,000 red blood cells per cu. mm., 7,250
white blood cells per cu. mm., and 5.2 grams of hemoglobin. On the
differential counts, in any one given trial, the largest difference
between initial and final counts on neutrophils was 16 cells, lympho-
cytes 15 cells, monocytes 6 cells, basophils 4 cells, and eosinOphils
5 cells.

Berkson et al., (1940) stated that following the usual

practice of taking twice the standard error as significant limits, the

J

erythrocyte count is date mine significantly wi thin :16 percent.

4..

Similarly, for a leurocyte coun t the count is determined signific-

antly within * 21 percent.

a

Each trial was further taoulated to show the trend

towards lowe n” and raising of the cellular constituents a1d hemo—

_J-‘O

globin content following the admilistration ofs ilfa :erasine. Re-

Virfl'vr

sults of this tab 01131131021 are shown in Table mum.

Table “in4. Blood Picture Following the “Ll‘hiStrdb’Oq of S ilfamera-

zine Cor pared with tie Initial Blood Iicture.

 

 

Test 1:0. of 130. showing Trials Tir als show; 111
Trials decrease showing idontica 1
increase results
REC 85 53 31 1
NBC 83 33 5O 2
Hemoglobin 85 53 22 10
% Neutrophils 69 42 23 4
% Lymphocytes 69 32 4 3
hi! 1 -' ...-L _ a 6 1 /
p monoeytes 9 12 41 10
i Basephils 69 7 33 29
fl Eosinophils 69 23 29 17

 

O

The results reveal a noticeable decrease in erythro-

cytcs, hemog looin content, and neutrOphils and an increase in ledwco-

cytes, monocytes, and bas0phils. In many of the trials the increase

or decrease was almost negligible.

file greatest variation in the red cell count obtained

‘

n an";" animal was 1:450 ,000; in tne majority there was very little

U

H-

.13-

change. There was a definite change in the white cell count. Small-
er doses gave no significant change but increased dosages of sulfamera-
zine produced a leukopenia. Hemoglobin changes were rather insig-
nificant except where larger doses were administered for several days
when a hemoglobinemia was produced; this varied as much as 5.2 grams
per 100 m1. of blood. Marked changes were found in the cellular
constituents as the dosage of sulfamerazine was increased. There

was a decrease in the percent of neutrophils, an increase in the per-
cent of lymphocytes and a slight increase in the percent of monocytes
as the dosage of the drug was increased. From these results it was
concluded that small doses of sulfamerazine were non-toxic but when
doses of two or more grains per pound of body weight were administered,
a definite tendency towards toxicity resulted. The range of blood
constituents in the latter trials was not found to be within the nor-
mal variations reported by Coffin (1945), halkamus (1944), and Boddie
(1946). Results of this tabulation are shown in Tables XXXVI and
XXXVII.

The oral route of administration was found to be the
one of choice for ease of administration. Either the tablet form,
powder form, or the emulsion may be used with equal ease. With
vicious dogs where placing the hand inside the dog's mouth is undes-
irable, the emulsion is preferred since it may be administered while
the dog is muzzled. The subcutaneous and intravenous routes of in-
jection are impractical with sodium sulfamerazine since large doses
are required and the dogs showed more objection to these routes when
given orally.

Only one dog exhibited any local or systemic reactions

.17) 331...... .4 .3. 3......434. 9.1.... 3.
«I44.
.H44I1.... qu‘IQD
r

 

-49-

 

i133... . 3. _. . ._ .
.-..» . r. r... .IrI...IFI .nlf. rush-3.”. o. ...w 1-0.44. 3.4.1.... 3.]. I. 1.. F ..r .I...Ir»
.I.. pf . [LE (Dru—I»? .8 0.1.... _ ._. n.1,}? II
... 8H,. .-H. be. 88.. Has.” flea
Oomfllhuoml F. Mn. 0+ ”Show. 30...... . 3.43.3 Cl 3 - )I.
..Ifi. ....\. 4 r .. ... . _.. r fl .PHH). nix... IO H .10 ...... ...
b.MHIb.mH «HNHI I.. on“. 00 .OIC ..n nooH HodH N: I c H-o_. .5...3 .HHH and. «“0398»; ..
I. ....x ..I \ .... IN 09 ... x C u U .... .. >
.A\‘ r' 3...; C : -— 'U .
H m5 Hdea oo.wkw.m o .2 0A, 2.... .. _. 93m ....H .HH
L Lh \ OOQH. N00 mNLOo : Z 2 = OOHKVQH. -r..H. MHH
.mII . . . \ x : z = 0020 w- I
. - w H SHAH $5-5... . .H NH.
m WillofiH +6 N o F I m UH ION 0
mos...” .I..H \ OHI...O O Hoilfim.» Now r H b m x. = : q.0 I
L .. rIO mH OoOHl3om. \3o3c o. ... \H Poms Swim . :6 F18. m. .. HHI
: _..m ... 0.3 To «HS __ _. .. 8.3 H QHH
LHI . I t x x z : 00-5
o\ \ MH O.O..Im0.mc OOoOIOoD o z r H OOH
W LHI Com-H OoHIHI 0 ff \ ... O mH N00 m 03
NL OH mq.0\'bo.fl O . \L \. .mw _ ... Q
00H. II? 0 \ I1 I m 01.. CI > ~ 3 2 mmHHlnp \. I. .- O
\ In L H.0rno.o n3..I3o.q -.hr H mH b.m z z a. .c s 30H
. .. 3 .... ...... . ...... u... 5.
_H..H..-.MH a. 3H,. me .. .. .. coco 8H
.HHIH..H .... 3.0-05 3H .6 . . .
0+ P 0OH'mNL0 MH NOOQ'O\ 0O 0.... O ”H O AQHJOOHHGJSOQHJO .3.
m. .310 2 H. 23,33. 3.. .... o _.H nu ...? A. .8 ,._. ..-V .. ....H 8H
boOHlmo? WN- olI. r mar. ,. f 0...“. no 0. OrdeafHH V : ...,H 03.
N. \H IMO MH'mmNII HOOIIm. n C l Nlfl Q‘s D AHHOIHI afiaj \r. \Olfi
3&5 « .. - m 0H H.3H .. .H. .V __ ...H omH
1.0% HH mb0 OI.“ “3.0n;|\.!0\. C Nu L 3m
IO \. U0 V. 0...: W. . AaHQAIK..rO3I.rV fivfl
o 1 NH A..M0O\V A) OHQU .04 = 3... NLHH
ms OHI \omVH 0?... NQMH \3o.\ tmofi H .v = ..HH mOH
9071' I ch h'\ I. 03-1
..MI O.NH ma oealmm.o OH mw.olio.3 O.MH 0.0H 5+.m Am:oo:d .oQI3V
M...IO ma no.0Ib. o e .¢Imo.h moua moo mboo A3«OQOsdHo.HHv = H NNH
..IJ. 04.1.. L. . . C. I r _
\0 \F O. OllmlA NL0.T|N~L 01.\ 10: \L00 OQO+N AHMOHQH 45.4.4. = H mNH
m mHIo.mH m. HI 0. ,o.muem. m ea 5.0 om.q 3 so V = a mHH
. . 0.3 . m... 2.5 Nye... V __ H ...HH
I I I \ 0.0.
0. I3; m. 3334 R5. 4 H V _. H 8H
0.0 3I I3.NH m.m2lm I. L a NH a 0.3H 0.0H +.3
o-H m.onm3.n .. b\ t Amdooss3soas 3
H. WHIE 9.-.. .... .I. .. . o ..H a... .... 3V __ HmH
\ Crl \ or «......oDIII on, L C n. O AQJHO HO... :6...) 4. .H.
A“. O..LFIIMO.\I.P. ANIOH lab. :1 (LC).\ 0r \. 0...; “DNA.- +\q.fl\d _ 4'.va = ..H. SN.NIfi
I.wIIo.3 - w o mo.mIH.m o.oI .c . AsoamHeszv a. 3
O ...f ..-. \H 0\_I 07k). (._.. O0 OH ......0 2 t
O h... 0 Cr A0D'H0 .I_0\ Nu\ m ....A )LJLMOI 3....
...: I . \L 0 .....H Ham. J. 0 Sr Pv = -..- mOH
Li. .3 .3... (H C a. J .r ..
(V r 03.03.03- L )H\_vd4.. J co .... I
.I p r C r» C. 02.2.. 0). -..: o. o A V n0 ..0 -r- “OH
.20 moquHJOHIJ mom -1. 1 ... C 0 Mom A03... HODH 1; I. II?
coangHsedlon. r . . c.;wc.. v .o;
.4 .r c c oQHnsI3idIIJ3 3 :
. ....r . ._. c 0.. doom MOO.

.moom Ha mo; :H mmbBon mooqm may mBHs ommdP Moo mszommzqumm
mo mOHBHMHmHmHmQ¢ -ra or nOAAOh mBAMDEHHmzoo MHASA So Mme o:H:on mvom :H mmmhon mooqm .HH>xMN mqm¢e
.Epom pmandp 2H mSHv @np mo coaponpmwnflfiuw mmpozmw *
.mafiamonwmoa I .m maflamommm I .m mmphooqoz I .2 mmvhoogmahq I .A mHHQQOHudoz I .2

 

-50_

 

HIO O mlH Hmlmm mowlm‘v. O O 0 cm #0 .x. = = ._moSHP mnm. ..VHH
NIH .vnnH +\lH wmlowm NOIMm O O H mm 00 * = = : mots. W...“ MHH
«no Nno Nno HonoN oonNo N H H mN mo * = = = mono «H NHH
HIO ._le mulo limoN owlbm O O O on «no * z .. ..moinnu. M H HHH
mno Hno Hno NmnoN oonNo o H o Hm mo * = = = 00H3p H oHH
mnN HnH onN omnHN monmo o o H mm oo * z a = oooo H ooH
mnN mnH HnH mmnNN monHo N H H wN mo * = = =mozHu m m. moH
Nno Nno HnH Hmno monmo o o o om Ho * = = = mOH:H H. ooH
mnN NnH NnH omnHm monom o H o Hm mo *mxoo m HHHoo mono H. ooH
H o N oN mo o o H mm oo Hmoooqoooopomv = MH MNH
mnH mno «no mNnoH oonmo N o o HN Ho Amooqm>oppoHv = HH oNH
H m m oN No N o o HN Ho AQOHmHoomv = MH oNH
H o N oN oo o o o om Ho Hpoozoov = HH oHH
NnH o o ONnHN monoo N o o HN Ho ApoHooHv = H. moH
H H o Nm mo m o H om Ho HmooogoHSOQva = H NNH
N N m oN Ho N H H o mo Hmoooo>ooooHv = H mNH
o N N ow oo o H o Hm mo AQOHmHoamV = H oHH
H H o mN on o H o Hm mo Homogemv = H oHH
mnH NnH o NmnMN oonmo N H H oN mo AHoHowov : H moH
mnH Hno mnH mNnoH monoo m o H on Ho Hmooonoosopomv z m. HNH
«no Nno mno NmnHN monoo N o o HN Ho Amooom>oppaHv = H. HNH
m o o mN No N o o «N Ho A:0HmHoomv = H- oHH
H H . m mN Ho m o H on Ho Homozoov = H. mHH
mnm o oHno HNnoH oonoo o o HH mH mo ApmHooHv mono H. NoH
m.m . u.m m.: 2.H m.m u.m m.m v.3 m.H m.m H.9H poo mcHonv .oH

nowpoowdmmIpmom EOHpoowonImnm onwndpmmdmafim ommmoq mom

-51-

following the administration of sulfamerazine. Anorexia and depress-
ion resulted from the oral administration of 4% grains (643.5 mg. per
kilogram) per pound of body weight daily in three divided doses for a

three day period.

-52 ..

SUNKARY AND CONCLUSIONS

The dogs exhibited the least objection to oral admin-
istration and the most objection to the intravenous route when sulfa-
merazine was injected. Sulfamerazine blood plasma levels following
oral administration were found to be slightly higher and persisted
for a longer period of time than those following the intravenous or
subcutaneous routes of administration. A concentration of 5 mg.
percent (therapeutic level) or higher was maintained for an average
of 20 hours following a single oral administration of 1 grain (143
mg. per kilogram) per pound of body weight, and 22 hours following
-§ grains (214.5 mg. per kilogram) per pound. Therapeutic blood
levels were not obtained for a sufficiently long period of time by
a single oral dosage of~§ grain (71.5 mg. per kilogram) per pound
of body weight in the dog.

Therapeutic blood levels of free sulfamerazine were
maintained over a prolonged period when given either subcutaneously
or intravenously only when l or 1% grains per pound of body weight
(1.1 and 1.6 ml. per pound respectively) were administered. These
two methods of administration were more objectionable to the dog
than the oral administration.

Sulfamerazine blood plasma levels of 5 mg. percent
or higher were maintained with the administration of-% grain (71.5
mg. per kilogram) per pound twice daily per os. Larger and/or
more frequent doses produced an unnecessarily high blood concentra-
tion.

There was no significant difference between the oral

use of sulfamerazine in the table H f rm, in powder form, or in an
emulsion in tiie dog as far as blood plas Ila levels were concei ned.
Studies of the blood picture inaicate that the firest-

est variation in the red cell co it ootainea '21 any antral was 3:

454,000; in the majority there was very " lo Cnan“e. Increased
dosages of Silfaxerazine during a three—day study produced a leuko-
penia in the normal healthy dog. Intraveno”; sly th e t;cnd wa sto-

wards a leukocytosis. Hem globi nemia was produced Hhen sulf amora-
z Lne was a~.fi.nisterw by mou .th at the rate of 4, frains (643. 5 mg.

per lcilO‘ram) per pound of body weight daily in three divided doses
for a three day period. WIi t1 the larger doses, the dose showed a

marked anore"ia annl depression whien las

(‘1'

3' .. *7 4- H
ed se/eral dans after the

_L

administration of the drug.

Albert

U}

, J. 0., and R. Graham. 1948. Sulfamerazine in the Treat-
ment of Fowl Cholera in Turkeys. Am. J. Vet. Res.

9: 310-313.

Anderson, C. W., J. C. Jones, J. B. Cooper, and C. L. Eorgan. 1947.
Sodium Sulfanerazine in the Treatment of Pollorum
Disease. J. A. V. M. i. 111: 141-143.

Berkson, J., T. D. Nagath, and K. Hurn. 1940. Error of Blood Cell

Count Kade With Hemocytoneter. an. J. Physiol. 128:

309-323.

Beyer, K. H., L. Peters, E. A. Patch, and H. F. Russo. 1944. The
Renal Elimination of Sulfamerazine, Sulfadiazine,
and Sulfathiazole by the Dog. J. Pharnacol. & Exper.
Theran. 8 : 239-246.

Beyer, K. H., H. F. Russo, E. A. Patch, L. Peters, and K. L. Sprague.
1946. The Formation an Excretion of Acetylated Sulfo-
nanides. J. Lab. and Clin. Zed. 31: 65-71.

Digler, J. 4., U. I. Clifton, and K. Werner. 1938. The Leukocyte
Response to Sulfonamide Therapi. J. A. K. A. 110:

Boddie, G. F. 1946. Diagnostic Methods in Veterinary Hedicine.
Oliver and Boyd Publishing Co., London. Blood of
Domestic Animals 320.

Bottorff, C. A., and J. S. Kiser. 1947. The Use of Sulfonamides
in the Control of Pullorum Disease. Poult. Sci.

26: 335-339.

Bratton, A. C., and E. K. Earshall, Jr. 1939. A New Coupling Compo-
nent for Sulfanilanide Determination. J. Biol. Chem.
128: 537.

Bryan A. H. 1946. Sulfamerazine in Veterinary Practice. J. A. V. K. A.

Clark, J. K., H. F. Flippen, and F. D. Hurphy. 1943. Studies on
2-su1fani1amido-4-methyl-pyrimidine (Sulfamerazine,
Sulfamethyldiazine) in Man. 11. Toxic Kanifesta-
tions. Am. J..Ked. Sci. 205: 846-851.

Coffin, D. L. 1945. Hanual of Veterinary Clinical Pathology. Com-
stock Publishing Co. Inc., Ithaca, H. Y. Erythrocyte
Enumeration 109-110; Leukocyte Differential 92-95;
Formal Ranges of Blood Cellular Elements 118.

-55-

Davis, D. D. 1943. The Binding of Sulfonamide Drugs by Plasma
Proteins. A Factor in Determining the Distribution
of Drugs in the Body. J, Clin. Investigation 22:
753.

Eads, F. E. 1949. The Clinical Use of Sulfamerazine in Infections
in Dogs and Cats. Ho. Amer. Vet. 30: 244-249.

Farr, H. K., and D. S. Jaquette. 1947. The Toxicity of Sulfamera—
zine to Chickens. Am. J. Vet. Res. 8: 216-220.

Favorite, G. 0., L. Reiner, and R. London. 1944. Acute Agranulo-
cytosis During Sulfamerazine Therapy. J. Lab. and
Clin. Fed. 29: 899-902.

Francis, J. 1947. Some Observations on the Pharmacology of the
Sulfonamides and 4:4éDiamino-dipheny1 Sulfone in
Domestic Animals. Vet. J. 103: 193.

Frisk, A. 3., G. Hagenan, S. Helander, and D. Sjogren. 1947.
"Sulpha—combination" - A Few Chemotherapeutic

Gilligan, D. R. 1943. Blood Levels of Sulfadiazine, Sulfamera—
merazine, and Sulfamethazine in Relation to Binding
in the Plasma. J. Pharmacol. & Exper. Therap. 79:
320-328.

Gordeuk, 3. Jr., and M. Learned. 1948. Sulfamerazine and Sulfa-
methazine Blood Levels in Turkeys. Poult. Sci. 27:

329-3 33 0

Green, D. F. 1944. Some Aspects of Sulfonamide Therapy. H. S. C.

Hageman, P. 0., C. G. Harford, S. S. Sobin, and R. E. Ahrens. 1943.
Sulfamerazine. A Clinical Study of its Pharmacodyb
namics, Therapeutic Value and Toxicity. J. A. K. A.

123: 325-330.

Hall, H. H., and U. N. Spink. 1943. Sulfamerazine. Clinical
Evaluation in 116 Cases. J. A. E. A. 123: 125-131.

Hoffman, W. S. 1941. Fhotelometric Clinical Chemistry. William
Morrow and Company, New York, N. Y. The Determination
of Sulfanilamide and its Derivatives. 214-225.

Jones, R. 1927. xalic Acid a Good White Cell Diluting Fluid.
J. Lab. and Clin. Zed. 12: 614.

Jennings, C. H., and G. Southwell-Sander. 1937. Anemia and Agranu-
locytosis During Sulfanilamide Therapy. Lancet 2:
899-901. ‘

'7

Kiser, J. S., M. h. Haters, H. I. Uirshup, and C. A. Eott torff. l
Sulfonamide Blood Concentrations in Chiczens and Tur-
key Poults. Poult. Sci. 27: 219-222

Kracke, R. R. 1947. Lefl gcytosis Due to Drugs. J. D. Lippincott
Co T11ii1alpbia. 181
O , - LA —-‘-~ A- 4—.

Langer, P. D., G. C. Poppensiek, C. R. Schroeder, R. L. Durkhart,
and Hark Uelsh. 1948. Sulfonamide Blood Concentra-

.1

tions in Ni k. Vet. Led. 4: 26-23.

Lehr, D. 1945. Inhibition of Drug Precipitation in the Urinary Tract
by the Use of Sulfonamide Kixtures. I. Sulfathiazole—
Sulfadiazine Hixture. Proc. Soc. EL er. Diol. and

;;§;. 53: 11-14.

Litchfield, J. T., Jr. 1948. As cited by No rthey. The Sulfonamides
and Allied Compounds. Heir nho 1d Publishing Corporation.
Few Iork, E. Y. Pharmaco lo ogyo of Sulfonamides and
Sulfone Drugs. 456-465.

Halha;us, D., and Th. Opperman. 1944. Clinical Diagnosis. Alex
Eger, Inc., Chicago. Formal Averages of Red Blood
Cells 288; Leukocytes 294.

An Evaluation of the Toxicity of Certain Sulfonamides
for YOL g Chickens. For t. Sci. 25: 59-64.

Lenten, H. L., and E. Graff. 1946. Blood Changes Related to Sulfon-
amide Therapy. II. Granulocytopenia Associated with
Use of Sulfonamides. Am. J. Ked. Sci. 211: 666-671.

Iurphy, F. D., J. K. Clark, and H. F. Flippen. 1943. Studies on
2—sulfani1amido-4-methyl-pyrimidine (Sulfamerizine,
Sulfamethyldiazine ) in Ian. I. Absorption, Distribu-
tion and Excretion. Am. J. Zed. Sci. 205: 717-726.

Peters, L., E. H.“ - eyer, and E. A. Pa c . 1944. The Renal Elimina-
tion of alia.ersz“ne (2-sulfanilamido— [-"etav1-ny1w.w-
dine) by the Dog. Federation Proc. 3: 1.

H. F. Flippen, A. H. Domm, J. J. Zimmerman, and L.
uh .arts. 1945. Rena Clearance of Salfamerazine,
.lf adi azine, Sulfathiazole, and Sulfapyridine in Man.
7311
.L

Reinhold, J. G.

rlfi

0’

S

S

J. harnacol. & Erner. Therap. 0): 279-237.

Sanford, A. K., C. Sheard, and A. E. Osterberg. 1933. The Photelo-
meter and its Use in the Clinical Laboratory. Am. J.
Clin. Path. 3: 412-414.

Scheidy, S. F., E. C. Icflanus, n. K. Tillson, and A. A. Pitt. 19 /.9.

Blood Concentrations Following the Administration of

F.)
0
<3
:1-
O
«2
8

n. ... 4-1. ".0 1.4.
ne and silfanetnasine to Catt

 

Seheid", S. F., and E. I. Tillson. 1947. Cm me1tratiou of Sulfa-
diasine, -1.11a:10M Mn mn 31113a1et11asiLe i1 the Llood
Of caUUle. tfs't. :::j..’ ’2: 232 -23Lr.

Schmidt, L. D., H. D. hu3hos, E. A. Dad 3or, and I. G. Schmidt. 19444
The Toxicity of Sulfanerazine and Sulfanethasine.
J. Pharmacol. & Exper. Therapy. 81: 17-42.

Severens, J. L., E. Roberts, and L. 3. Card. 1945. The Effect of

‘1

Sulfonanides in 33d1cin3 Vorwtalit ;ron Pallorun Dis-
eas in the Domestic -onl. Dealt. Sci. 24: 155-158.

 

Sieber, R. H., and I. Clark. 1945. Factors Influenc 33 the hainten-
ance of Blood Levels of Sulfonaniees. A:. J. Zed. Sci.

209: 813.

Thorp, U. T. S., S. Sorde‘h, Jr., P. J. Slants, and K. Learned. 1947.
The Che :other apy of Cecal Coco di osis. An. J. Vet.

 

Des. 8: 196-20 .

Todd, J. C., am A. II. Sanford. 1943. Clinical Di a311osis by Labor-
atory'Iethods. U. A. Saunders Co., -hiladclgthia.
DiL 1ti n3 Fluid of Leake and Guy 246. Hri3ht's Stain
253.

Uastrach, H. 3., and G. Teu’s 194 A Clinic 1 Lvaliat ion of Sulfa-

_J
merazine in Infections of Cattle. J. A. V. X. A. 110:
103-112

Ueleh, A. D., P. A. Mattis, E. S. Keelle, and A. R. Latven. 1944.
Sulfanerazine. II. Sdlfonanid e Concentrations in
the Blood of Kan Produced by Small, Daily, 01al Doses
of Sulfamerazine, Sulfanethazine, Sulfadiazine, and
Sulfa thiazole. In. J. Zed. Sci. 208: 187-192.

Helch, A. D., P. A. Hattis, A. R. Latven, U. E. Benson, end E. H.
Shiels. 1‘43. Sdlfanerazine (2-sulfanila.LTdO-4-
nethyl—py imi ins). I. A Comparison of Salfa.era-
zine with Sulfadiazine on the Basis of Absorption,
Excretion, and Toxicity. J. Pharnacol & Exper.
Therap. 77: 357-391-

.Jelsh, 3., P. h. Lan3er, R. L. Durkhart, C. R. Schroeder, and G. C.
Peppensieh. 1943. Sulfonanide Concentrations in

“'1

2 xes. Vet.1ei. 43: 522-524.

Welsh, H., C. E. Schroeder, D. F. Vronan, L. eddin, R. L. Durkhart,
and P. Lan3cr. 1946. The Fate of-r Gr. /Lo. Body
Height of 7 Sulfonanides in 7 A11ina1 Species. Tree.
U. S. Livestock Sanitary. ' sm‘1., Chica 3'0, 50th

mual Iieeting: 213-234.

mafigmoaflmom I .m mafinmomwm I .m mmphooqo: I A: mmphoogmahq I .q mHH390Hpsoz I .z
woOHn H0 .H8 OOH amm aHQOHmosmg mo means I .H5 OOH\.m .pm

mflamo cooam mpfln3.I .o.m.:

mHHmo wooap dam I .o.m.m

wamwam wooan mo .HE OOH Ham mnfinmnoawmadm mo mawhmwfiafiz I w .ma oaflnwpmswmadm

 

 

N mH om 5H ooa.HH ooo.oo>.m m.o o.H .9; Hm
H 0H om m.oH ooo.mH ooo.ooo.m 0.0 o.m .Hg NH ,
o.m m.m .ng m
H.m m.m .nn 5
m.m .q.m .HH Q
o.m q.m .pn m
m H 0H mp m.mH ooq.OH ooo.o¢m.m o.m ¢.m .9: q
H.m m.m .9H m
>.H o.m .pg m
m.H @.H .HH H
HwHSQSp .80 H
om om OHA 01H .2 ...—“E OOH\OM oUomoB oUomom R ems HfizmpGH
HaHpsopmHHHm .nm onHanoSQHHsm osHB

 

 

mmHUSHm wooam

hdpawmn hapcmnwmm¢ I AHOpmHm

0:0 I mmmw .om mono I mww Hog mmawa pnmwoz Avon .na\.pm m.l mmoo

.mp» m .mnH mm oHdz HpoHnaav Hugo I mHSOm

mH\H\m HOH .oz mom no m:0praHanopoa NH mHHwe

O OH Om OO O.HH OOO.NH OOO.OOm.O 0.0 O.H .HO Hm

 

O.H O.m .HO NH
O O OH OO m.mH OOO.mH OOO.OOH.O O.m O.m .HO O
q.m N.m .HO O
O.m m.m .HO O
O70. Tm ..E m
m O Hm OO mH OOO.NH OOOIOOO.O m.m q.m .HO O
M» O.m O.m .9: m
. O.H m.m .OO O
O.H O.H .pg H
nstnsH .50 H
.m .m .2 .H .2 .Ha OOH\.m .O.m.z .O.m.m m .ma HO>HOHOH
HOHpsogoOHHO .Om OOHNOOOOOOHOO OaHH

 

mowfifim @0on

533% Epsopwmmd I bopmflm

one I 32% .02 8:0 I saw .me momma pgmfloz I303 .OH\..Hm .m I mmom

.mgh m .mOH Om OHO: AHOHOOHV HOHO I wagon

OH\OH\M mOH .oz MOO no OOOHHOOHEHOHOO M OHOOO

-60..

 

 

O.H O.O .OO OH
N Om OO m.mH OON.OH OOO.OOO.O O.N N.m .9: HN
H ON ON mH OON.O OOO.OHO.O O.N N.q .HO NH
m.m H.m ..E m
H N Nm no mH OO0.0 OO0.000.0 O.H O.m .n: O
H.m N.O .HO O
O.m H.O .Hg m
ON NO OH OOO.O OOO.mOm.O O.H m.O .pa H
O.H m.m ..E m
m.m O.m .HO N
m H HN mp mH OOH.OH OOO.OmN.o O.H O. .HO H
NOHOOOO .ao H
.O .O .2 .H .2 .Ha OOH\.m .O.m.: .O.O.O m .ms Happmng
HOHpgogommHO .nm mzHNOHoEOMHdm SHE

 

0:0 I mth .03

.upm m .nOH OO

33

ooao I haw pom mmsHm
OHOE

MOH .ou mom no mGOHpOQHfipOpom

P

OOHwfipm OOOHm
AAIHOO: HHpmoHOmmd I MHOpmHN
pgmHm: Avon .nH\.Hm H I onoa

AHOHQOHV HOMO I madcm

4m. GHQdB

kr

 

 

 

N ON OO O Omm.NH OOO.OmO.m O.m O.m .HO ON
O3 0.0 ..3 NH
m.m H.O .OO O
ON HO O.OH OOO.HH OOO.OOO.O O.m H.O .HO O
O.m H.O .OO O
HO H.O ..E m
H ON HO O.OH OOH.HH OOO.OHO.O O.O O.O .HO O
.I.. Oom Nd 33 m
O ,2 H.O ..E N
O.H O.H ..HHH H
.HHOHdOHHS 2.8 H
.m .O .O .H .3 .Ha OOH\.O .O.O.: .O.O.m m .Oa HOOOOOOH
HdeHHOHDMMH .. .013. Omdndhowamaflm 93,.“
OOHOOOO OOOHO
bfiHOoHH 5320.39an I .9333
98 I O.H..OHO 6.... 028 I BOO .HOOH OOOHO ..EmHo: Avon .OHH\..Hm ......H I omom
.noa OH .OOH ON oHOz AOOHOOOV HOHO I Opsom

mH\o\O OOH .0; mom :0 OOOdezngopom HHH oHOOO

 

 

 

N O OO N.OH OOO.HH OOO.OON.O O.N O.m .OO ON
O.m O.H .OO NH
NO O.H .3 O
ON OO N.OH OOO.HH OOO.ONH.O O.H O.m .OO O
O.m 0.0 .OO O
O.“ O.m .OO O
H ON OO O.mH OOO.OH OOO.OON.O O.m O.m .OO O
LO O.m O.H .nO m
Km” 0 0
O.N O m pg N
O.H O.H .OO H
.HHOHdQHS .fio H
.m .m .m .H .3 .He OOH\.O .O.O.: .O.O.m O .OO HOOHOOOH
HOHQQOMOMQHO . m oHHHu.O.H2HO.HHHHm mafia
OOHOOOO OOOHO
prHOOHH \HHonHOHEO‘ I eroOanm
one I mth .0: mono I .96 mom mofiflu pmwwoz x309. .O.H_\..Hm NH I omom
.Oos OH .OOH ON oHO: HOOHOOOV HOOO I mpsom
O+\\o\m mOH .on mom no OGOHQOHHUOHBQO HHHM OHQOH

 

 

 

O H m ON OO 0.0H OON.HH OOO.OOH.O N.N O.N .OO ON
m.m m.N .nn NH
O.H NO .2 O
O.m O.OH ..E O
O.OH OOO.OH OOO.OON.O O.H N.H .OO H
QOHSQSO .fio H
.N .O .u .H .O .HO OOH\.O .O.m.: .O .O .m O .OO HOOOOOOH
HdflpmomOMMHO .9: OQHNOHOEOMHOO mEHe
OOHOSOO O00Hm
rmpldmm thsopdmmd I huopmHm
000 I uth .0: 0000 I haw hog mmEHB paMH0:.hdon .QH\.pm m.I 0m0O
.mog mH .OnH mm 0HO3 Apoczomv HOHO I opsom
mw\Om\w mHH .0: MOO 00 mQOdeGHEhopOO >HH QHQOB

 

 

H H ON OO O.OH OOO.O OOO.OOOOO 0.0 0.0 .OO ON
O.m m.m .OO NH
moo NCO OOHflH m
0.0 0.0 .OO O
0.0H OO0.0 OOO.OOO.O H.m O.N .OO H
.
In... ll
x0
_
anfimdp .Eo H
.O .O .: .H .2 .Hs OOH\.O .O.O.: .O.O.m m .ma HOOOOOOH
.Qm OQHNOHOSOOHOO maHH

HOHOOOOOOOHO

O00 I OOOO .0:
.mhm mm

OO\OO\O

.mpH Om

@000 I haw pwm moSHO
meaom

OHH .om mom HH0 SOprcflfihopom

OOHOSpm OOOHm

hmpHOm: thsondmm< I OHOOOHm
pamHoz hwoa .QH\.pm H I mmom
mpSOm

Apowzomv prO I

.M OHQOH

-65—

 

 

H N ON OO 0.0H OO0.0H OOO.OOHOO m.m O.m .OO ON
N.O 0.0 .OO NH
N.O H.O .OO O
0.0 N.O .OO O
0.0H Omm.NH OOO.OOO.O m.m m.m .OO H
andflsp .50 H
.O .O .3 .H .2 .HO OOH\.O .O.O.z .O.O.O O .OO HOOOOOOH
Hdeu0p0OMHO .pm 00HN690§OMHdm ofiHB
OOOOOOO OOOHO
hngmmg thmonOmmd I muopmHm
000 I mmOO .03 0000 I AOO Mom OOEHH pmeoz MOOQ .QH\.gm MH I mmoa
.OOO m .OOH mmm OHO: AOOOOOOO HOOO I OOOOO
OO\OO\O OHH .om OOO :0 OOOOOOOOOOOOO HON OHOOO

 

 

 

m ON NO O.NH OOOONH OOO.OONOO O.H O.H .OO ON
O.H O.N .HA NH
H.O 0.0 .9: w
O.O 3 .5 O
H.OH OOOONH OOOOONH.O O.H 0.0 .OO H
HOH595p .50 H
.O .O .O .H .2 .He OOH\.O .O.m.: .O.O.O O .OO HOOOOOOH
HOOOQOOOOOHO .Qm 00HOOH0EOMHdm 0EHH
00OO5pm Ooon
hanwog thc090mmm I hhopmHm
000 I thO .03 0000 I OOO pom m0EHB pamfl03.hdon .nH\.pm m.I 000m
.005 mH .mQH om 0H0; AQOHOHOEHV HOMO I 0p50m

OO\ON\O

mHH .om mom :0 QOHOOQHSO0P0O

HH>N 0HQOH

 

 

 

N N OO OO 0.0H OOO.O OOO.OOH.O 0.0 0.0 ..3 ON
H.O 0.0 .M: NH
0.0 o.m .Mg m
m.O N.O .Mg O
H.OH OOOOO OOO.OOO.O O.N O.N ..E H
. MOHSQOp a0 H
.m .0 .2 .H .O .HO OOH\..O .90.: .OOO O .OO HOEOOOH
HOHp00M0OmHO .pm 00HOOM0SOMH5m 0&H9
O0HOSPm O00Hm
hngwma OHpmmpmeO I hMOpmHm
0:0 I name .0: 0000 I haw Mmm OOEOB pmMO03_%O0£ .nH\.Mm H I 000O
.OMA mm .OQH MOM 0Hwa0m AnoHOHSEmv HOMO I 0p50m

OO\MN\O

OHH .02 MOO 00 QOOPOGHEMopom

HHH>N 0HQOB

 

 

H m m ON OO O.OH OOO.HH OOOOOOOOO 0.0 0.0 .OO ON
0.0 0.0 .Mg NH
N.O m.o .Mg m
N.m m.b .Mg O
OH OOm.NH OOO.OOH.O 0.0 0.0 .OO H
_
Om MOHdndp .50 H
.O .O .3 .H .O .HO OOH\.O .O.O.3 .O.O.O O .OO HOOOOOOH
H0H¢00~0OOHO .Qm 0GHN0M050dem 0EHB
m0fiO5pm O00Hm
hngO0g ths0M0mmd I hMopmflm
000 I OAOO .05 0000 I mdw Mom m0fifia pamw03 OOOQ .QH\.Mm m. I 000O
.moa mH .mnH mam 0H0: AQOOOHOEMV HOMO I 0u00m
OO\H\O ONH 63 OOO no OOSOOOEBOO OHH OHOOO

 

H H ON OO O.OH OOH.NH OOO.OOO.O H.H N.O .OO ON

O.N O.H .OO NH

0.0 H.N .0: O
H O OH OO 0.0H OOm.HH OOO.OOO.O 0.0 0.0 .OO O
N H N ON NO O.OH OOO.HH OOO.ONHOO O.NH O.NH .OO O
m H ON OO 0.0H OOO.OH OOO.OOO.O O.HH O.OH .OO H

 

-69-

.HOHHfisv . 50 H

.O .m .0 .H .2 .Ha OOH\.O .0.0.z .0.0.0 O .OO HOOOOOOH
H0300M0mmfi O .02 00H00M0EIOMHdm 05m.

 

O0Hgfim 00on
5.03.000 th00MOmm4 I MMopOfl:
000 I 05.00 .0: 0000 I WOO Mom 0083. 90mg: 0300. .OrH\.M.m~ ._m I 000m

.008 mH .OQH mm 0H03 0:0000p0090m I 00.00er

OO\OH\O HNH .00 OOO no OOOOOOOOOOOOO 00 OHOOO

 

O.H O.H .M0 wv

 

O.N O.N .OO OO
O H NO OO 0.0H OOO.OH OOO.OOO.O 0.0 0.0 .OO ON
0.0H H.OH .0: NH
0.0H 0.0H .OO O
N.OH N.OH .OO O
0.0H 0.0H .00 m
0.0H OON.mH OO0.000.0 0.0H 0.0H .00 H
.
mW OOHOOOO .Oo H
.0 .O .0 .H .2 .HO OOH\.O .O.m.: .O.O.O O .OO HOOOOOOH
H0Hp0000mmHO .nm 00H00M0009H5m maHB

 

000 I OAOO .0:
.000 mH .OQH mm
OO\ON\O

0000 I Add 00% m00fle

0H02

00w009m OOOHm
hanmmn th00M0mm¢ I hMOpmHm
OOOOOO OOOO .OH\.OO H I OOOO

000000p0030m I 0p00m

NNH .OO OOO no OOOOOOOOOOOOO HON OHOOO

 

 

 

0.0 m.m .00 OO
w.m . b.m .M0 cm
H N ON OO H.OH OOO.NH OOO.OOO.O O.HH 0.0H .0: ON
N.OH 0.0H .M0 NH
N.OH N.OH .M0 m
m.HN O.Hm .M0 m
N.NN O.NN .00 m
m0 N.OH OOO.NH OOO.OOOOO 0.0H 0.0H .00 H
.
MOHSQSP .00 H
.O .0 .0 .H .0 .Ha OOH\.O .O.O.: .O.O.O O .OO HOOOOOOH
H0Hp00M09OHO .pm 00O0000009H0m 00HB
00H009m Ooon
h0pH000 me00M00OO I hMopmHm
000 I OAOO .0: 0000 I 506 00g 000H9 p0mH03.hwon .0H\.0m MH I 0000
.th m .09H OO 0H02 00000090000m I opsom

OO\O N\O

MNH .02 mom 00 m00HpO0HEM0p0O

-72-

 

 

 

H H ON HO O.NH OOO.OH OOO.OOOOO O.N O.H .00 ON
O N O HN OO N.OH OOO.OH OOOOOON.O 0.0 N.O .00 NH
0.0 O.m .00 0
0.0 0.0 .M0 O
O.NH OOO.OH OOO.OOO.O 0.0 0.0 .00 O
H N N NN OO O.NH OONOOH OOO.OOOOO O.OH N.O .OO N
H NO OO 0.0H OOH.OH OOO.ONO.O N.HH 0.0 .00 H
0.0H O.OH .00 O
MOHSQSp .50 H
.0 .0 .0 .H .2 .HO OOH\.O .O.O.z .O.O.O O .OO HOOOOOOH
HOOOOOOOOOOO .OO OOOOOOOOOOHOO @000
OOOOOOO OOOHO
hflpH000 th0000mm¢ I hMopmHm
000 I 0h00 .02 0000 I 500 M09 00EHB p0MH03 Avon .QH\¢Mm m.I 000G
.000 mH . .mnH ON 0H0; 00000>0Mp0H I opfiom

OO\OH\O

ONH .om mom 00 000HOO0OEM090O

HHHMN 0HQOH

-73-

 

O.H O.H .00 m0

 

 

O.m O.m .00 On
N N O ON OO O.NH OOH.HH OO0.0NN.O 0.0 0.0 .00 ON
0.0 0.0 .00 NH
0.0 0.0 .00 O
O.HH 0.0 .00 O
0.0H H.HH .00 m
O.NH OOO.OH OOO.OOO.O H.OH O.NH .00 H
O.NN O.HN .00 O
MOHSQSP .50 H
.0 .0 .0 .H .0 .Ha OOH\.O .O.0.: .O.0.O O .00 HO>0OOOH
H00000000000 .00 0000000000HOO 0000
0000000 OOOHO
h0pH000 me00M00O0 I hMopmHm
000 I 0500 .0m 0000 I 500 000 000HB @0MH03 0000 .0H\.0m H I 000O
.00» m .00H mmm 0H0: 00000>00p0H I 0p00m
OO\ON\O ONH .oz 000 so 00000000000000 OHON OHOOO

-74-

 

 

 

o.m o.m ..E m...
o.m 0.0 .00 ._m
N m 0 0N 00 0.00 000.00 000.0No.0 0.00 0.00 .00 0N
m N m 0N 00 m.N0 000.00 000.000.0 0.00 0.00 .00 NH
0 N 00 m0 0.m0 000.00 ooo.oou.0 0.00 0.00 .00 m
N 0 0N 00 0.00 000.00 ooo.omm.0 N.00 0.00 .00 0
H64 m}: .00. m
m.om H.ON .00 H
N.mm H.N..m .00 o
00d..03...0 20.0 H
.m .0 .u .0 .0 .00 000\.m .0.0.0 .0.0.0 u 00000000
H0000000MMMQ .Qm 000000000mafim mafia
I: mowUfipm wooam
3.0.00.0 5.300000%...0 I 500000.00.
000 I 0.0.00, .0.” 0000 I 000.00 009 00.0 0.0%....0: 0.0.002 n~H\.0m ...”..m I 000m
.000 ma .uga ”mm 15; 00000>0000H I 00:0m
mq\bm m. omfl .on .000. 0.0 m:0.....000fl..00 09 Run 3000.

.mfipw 039 MO QOdeppmficflEdd “mapoaw mmpdowwafl *

 

 

 

m mm no H.¢H OOHIm ooo.o®o.m 0.0 m.o .na mu
H.b o.m .93 oo
0.0 v.0 .pg om
O.OH «.0 .pg mm
q N m pm mo N.ma oom.ma ooo.ou¢.o 0.0 o.m *.aa mq
H.u m.> .g: cm
H.w 0.0 .gs mm
m.b H.m .H: mm
m m «m mo m.mH oom.HH oooaomm.o o.m m.m $.pg «m
o.m 0.5 .aa NH
m.» o.m .ug m
0.5 q.u .a: <
m.m o.m .pg a
pmfldDSPxfifio H
.m .m .2 .q .3 .Ha ooa\.m .o.m.z .o.m.m n .ma prpoan
Hdflpsopommam .3: ouflmwaoummaflm mafia
moflcnpm cooam
hmpfimma hapcmpwmmd I muopmflm
oohflp I mhdw .0: mono I Haw pom mofifle pnmflms Avon .Qfl\.nm me I mmom
.mgm m .mpH um mam: Apofinmyv Hake I opsom

m¢\mfi\w

NHH .om mom :0 macapmcflahmpom

HHH>HN mHQmH

.deU 039 mo soapdppmflswfidd moguoqd nopdoadfiw *

 

 

 

N N m on mo N.mH coo.OH 000.00N.o O.N 0.0 .pg Nb

m.N N.N .Ng ow

m.m O.H. ..E «6

0.0 H.N .ng Nm
N N m mN mo «.mH OONIOH coo“oomae H.m N.N $.NH NH
N.O N.m .Ha om
O.N «.0 .pa Nm
0.0 H.@ .pg NN
,w H H , o HN mo N.mH coo.OH ooo“omq~o O.H m.m $.pg «N
4 H.H ma ..2 NH
O.N. moo O.HQ «w

m H N om Ho m.OH oomgo ooo.o>m“© m.o N.N .Ng H

H.N O.N .NH H

1‘ . .Hw-mH,HnHH\,p 5.0 H
.u .m .u .H .H .Ha OOH\:N .o.m.z .o.m.m n .ma Hw>popaH

Hmwpqonmmmwn .Qm mzwmwhmfldmajn mflfia

 

 

0093p I mamv .om

5:. m .2: 3

mH\HH\m

@030 I Maw Hag moawn

oHduH

OOH .oz mom so meowpdmflShmpmm

mchspm cooflm

Aspadoz hfipsmpmmm¢ I hhopmfiu
pmmwoz hwoa .,H\.Hm H I mmom
HHoHpmev Hugo I mHsom

HH>MH oHQdH

-79-

.nfipd esp mo QOprhpmHQHde Hmmwocd moudomunfl *

 

 

 

m N N 5N No m.mH oom.o coo.QMN.< O.NN N.NN .HH Nb
0.0m H.Nm .pa oo
m.oN N.NN *.Hg on
0.0N O.NN .pa Nm

N H om no O.NH 00¢.OH 000.0Hm.¢ N.NN o.mN $.HH NH
m.oN N.NN .H: on
O.NN m.MN $.Hg Nm
0.0H «.0H .Ha m

H H N Nm No O.NH omo.m ooo.omm.q N.NH N.NH *.Hg «N
O.NH N.NH .a: NH
v.0 Nob *ohx m
m.< N.N .pg q
m.N o.m .aa H

hmHfiflfip .fio H
.m .m .u .H .: .He OOH\.N .o.m.z .o.m.m m Hd>gmHnH
HNHHaoNOHHHQ . m oaHndgogdHHfim maHm
mmespm NOOHm
hngwmg thconggd I hpopmflw
mega» I mhww .oz 039 I haw Ham moEHH pQMHmS 5609 .nH\.Hm H I mwom
.mhh MN .mflH 0% deEom ApmHfldBV Hwho I mPSOK

NN\0H\N

OHH .ou mmom no mQOprcHEHmme

HNH mHQwB

.20-

.mzpd may 90 mOdeHpmHmszw heapomd mmpNOHduH .

s
1"

 

 

m N H 3 mm H.HH 8N5 0863.6 mam Won ..8. NH.
0.0 m.mm .Hg 00
m.<m m.mm $.pn om
m.qM 0.0m .9: mm

H H N Hm No HRH RiHH 80.83. 0.3 mém .....E 3
N.mm v.0m .9: on
O.NN m.oN $.Hg mm
O.NN ¢.<N .Hg mm

H H H mm R HRH 03.2 08.036 H.NH 0.3 .....5 ...N
o.mH H.5H .9: NH
m.OH 0.0H *.H£ m
HOMW 00m O.HHM 4V.
o.m m.m .Hn H

.HHNHHHQHHHH . E H
.m .m .u .H .H .HE 8H\.N .05.: .u.m.m m .me 350ch
HNHHQNHOMQHQ .2: quNwHQHNMHsn oEHH
mmesvm GOOHm
hngdog thcoHdmm¢_I huopmHm
@0939 I mmww .0: 039 I 566 pom mofiHm p:MHo: thQ .QH\.pm m. I omom
mg» m .an 0% dem ApOHnwev Hapo I mason

3\NH\N

MHH

.0: mom no mQOdecHSHome

HMHH mHQdB

AJ - I I II .I 4.. II. III I. I II III
.. HSHS 0H5. MO $0 _.>.. 91%., H _.HH T, g H.H). «HUANG 03D nOu.H...O v.32... _r
l b

a. O .0 rr. 0 f o

)4 . f. )q . ) . N... . I
H HN : 0.. 0m: . o o QNNN . O.NN N.o\ .N: o

. _.Nfi N.NQ .HH 00
O l- ..I O _I
m Om N.ON * hr 0m

0.nm O.HM N.N: mm

 

 

H N 0N ON .HH NON.N 000.00N.N N.N N.NN N.Na NN
.mm m.0m .Hm wm
O.NN O.Jm *..£ mm
o.mm o.%m .HH mm
)‘ '11 h A Q ,I I L. .r‘
H m m 00 n.0r 000 w 000 000 o O.NN ..Nm x. H \0
II. “ R Q I1 44. 2
b.mr 000 b 000 000 m m.or m.bH .H: 0H
« a a
b.MH OOH m 000 one m o.mH m.OH *.H£ m
. . I. I N I N ,. +
O.HH omr Hr 000 000 m 0.0H «.m *.p: \
N.NH 000.HH 000.0Nm.m N.N. N.N .NN H
cHd.m.,....r....,.w -0 H
.m .m .m .H .H .H5 00H\.N .N.N .0.m.m a .Na HN>Nong
N-HHfi no @003
proz prsogdc:H I 5H0p0.;
oonzp I n>53 .0; oopdN 00 Mom uofiHn pH...H0 3» H003 .QH\. hm m. I 0m0H
.Nah m .NQH mhq 0Hdu AHNHQNMV cho I opdou

‘xi'

mfi mm m fiHH .ou tom m0 NuOHp.0:NHm0pmm HH:5% OHQNH

T AA ‘.m-*‘I—‘ 'r‘v V‘f .F‘fi T‘jf‘? Q‘ A- 7-...- .1 {'1'?“ q \s ..-—_.r‘ rflH - rj—fiuw
I

:HLV SJ-JAJ-W con-o! .LJVUD J‘- \rh—JAJVoO-nl-biu ‘ig—J ‘g—J-...-..I.__0JL— goLLU‘I LI.L‘

SJE "fi7h24"‘

-‘- \n-tr‘ 1" w (10"1fio _n ‘ n d¢q '6 "n“."xh
_o—quQ ...O~ “vq_ V O— - hat-v v r O— IJV—v--CC
. _.c‘ ‘K, ._ fi_-_ ‘ J‘n ,—
...LC'...LUQ..L 3 UK... U0 C1011»:
7‘1 "1 ,
;‘ 0 Ad. .LJGLAL)
1949
.‘ .‘ 1 J—
An “esprect
r1‘1 I 'nnmq -\ ":1fi0 ‘rujm 11.1“": "1% (...-'—H\..4.N,.1 ~ -v- ‘rhfi"? ‘v‘lf‘ -1‘v1"‘qfl "IT' “1
NJ _. Lb.&bl 11.4“.-.29 .49..“ 5‘.‘-N.bi-.‘.-—_p JV.— a,“ DO v \‘M— ._OH-uJ L u'UL-ILJ W vb

I 0 'OH '1' rlr‘J‘fiv” ":‘ l ‘ '3'“n 7 3“":7 " ‘6 S‘s, fl ‘ fiver-'31" n.
CLO“, ...—’9.) U0 “v. UV A....'_ 110 U:.\/ ‘JlOOi If -..... .J . .Cn. 4-9 ‘h‘ L.,...) 0... fl I..."ch n..- “5.1.44 0 {...-El
-‘ -. n n 'l A. I“ 7 fl -? --\ ‘C . ’1‘ ...: N . ‘~ . J‘ '1“, J,“ - K "‘
em--erent lnt r’el: lollowlLQ 1 s eNL;ll:.ration. (2) one freulenod

blood plucra lecels, and (3) the effect 01 the peripheral blood. -10
nunflred forty-six s: fa 3333118 trials were com; eted sin; 7 normal
healthy dogs.

FoL‘ovir' the sin;le oral administration 03-2, 1, and

fl . 4 «I. r‘ ‘4 . h ‘ Y. a a Q n _\ 0 ‘V‘ o I n‘ a 7 1 ‘ a
. _ r - p - fl r‘q~ Q « I n v \ t‘ fi if -‘ .fi -‘ -\ N ‘1, ‘ ‘ \ ~ - . w _ f -- _ I v‘ .A- '.u
l ' Dab uLn...) O: ”2‘...— “.34.. -..—J_-LLV ch... LH'OVJLKA 0.1. :JVu.JI IIJ“ 1L; U Ll..-\../ Lin-.00: 1.;— 5)....“

I'd

Cnflficnrtnn'l‘: w 1-r-r‘ {‘OHwJ
VA- V A ‘LAU... .- V‘Av‘

-- I‘MVH

4‘ 1_ na* ~- -ar ---. a 'L I -* J“ A n .. q - ~- -- ’ -
' . 3 ? f ‘. I > ; fifi ‘,
UO I 3 “‘H .— v)n;o‘-r1-UO ”I UJIV 5.5:}...v Itfi*v11 .‘_I bi]. ._n

tablet 3 :2, in powder form or in en enmlsion

Sllfcterezine blood elesne levels follow‘n; oral ai-
mlnlstreclon were found to be 311"
lon er oerloi of tige than those fell r’r' the intr;"enoxs or SLbcut—

. ' .. - .2 .- n - 4-: ~ ~ , .I.
allCOJJCJ TOLI‘QC S 0:. 61.5.2111... orlulo-.. n C.)11C31.L»l"€i. 02.011 03. 9 I19. liCI‘CCI‘lu

(J

(.“e-ade1t 0 level) or higher was melnzeino for an average 0 2O

1. .. .4 ..° .. .9 . 1.: .2 1.. 4n n l ,.- : '1 If) .. .

no. lOlthan a olnule oral auufiL;3u::Ulon 01 Uruln (_NJ ma. oer
1"'33. 'Tin‘r‘.‘ '1“ ‘-. "3 1 f" 1‘F\l".-r '31—: 3.1“: q‘fi1 ’3") n" 1n 1 Vv-u‘—l :l‘- “‘r.:\ -‘
A-.. Qua. (.5..-) i VJ. "_.'O wi1‘-'. O_ U'ssd 1" "VJ—J..- U , wAkL 5...». 11V ..le 1‘01 0” .4 LJ'L 9.4-1.)

'3 :' .-.. - '--1 .- -,,. -- . x1 -- .44 a .. ,- - l ,
Nlfl.) NU. per “1 ogre ) Lu? Oind. .gere;e;u_ olooN lewe s were

Abstract cont'd - F. 3. Sade

_L 1‘1. 0 _ 1 31 _ n‘nf -. “‘1 ‘_ ‘N‘" V __ 3 T .11 1‘3‘ _f .'- ,‘1
lob O'UQlMBL lor a SllllclelulJ loaq gerloa or olae b, a S_”ULGO m‘al
n .. n " ,.- -: . .r: -. a: .- ‘ ..- , ., o r ,.‘-.. .° J-
aosaue ol'; y‘aln (71.; m¢. per allouxla per yoand o: aoaJ welbav

r-n. . a - _... ,. .. :- 3" a- M n .2. .. -
leeragoaulc blood levels ol lree galfarerazrae here

.. '- - ' .. - ,. ._-. .. a -3 . j - “.2. - no. .- . -
galnoalnei ower a yrolenued ,erlod ween place eluae: saeclo.1eoas j

 

 

 

 

.,
.. .‘J-q ~ v q -<. -— - .1qu 7-, I. - l _ r - - 7‘1 (3 '2 -"J‘
or lne‘axeaoaslj eel; uaea l or a 'rain: per poaaa or bod' Helga»
, a
l 1 '1 j 1 I" 7‘1 “ 3“ “x 1 "r1 “A“.H""nr'\"‘ Tfi'fiq ' .'.:x 14:) - ' <4 ’f\“1)4‘| r“ ”a“ ‘3
. all.» .I.. J -4... ina. Mogul»; ...~ g 19v w... L, "d l v... 'v Lug- .'.—.l...) u\., we; 0 .'.-~\nh)\./
"’ " v
J'V “"\’!‘-" 1‘ a f) r‘ 1 ‘. ‘: fl'l' ‘Ar‘ J”: “‘1 ' ’{3‘1 '\ ".'I 74/3 ‘ . r“ F} J‘, -'- 110 fi 7. X;
mIO ..'.'\./ U-LO(«.Q 0.1. U.»L..\-’.31.L.._a Lia. .. ia.—Oi; {39.1. ‘v’ “LO; ‘4 OsJJ LC"- ‘1“ {01-3. 31."; L13 (Janna L OD "‘
_- .L‘ ,::... - 4.- \ -‘
wean UJC oral aan_n:* "n+eon
H‘s-1 9": ~<~' - "1‘. 1“- I“ ~ -! v '0 :T‘ V‘ ‘ ’ L
allraheraZLMe ol00a Llas: a levels ol 2 J3. oeloeau

‘ 'I‘ 1
‘~ ,. (3“ ~ "av-r.“ ?"‘"1 at w' 1". n-w 1.," "‘ "3 q; ‘ v17" ' "J' WHJ‘ 0"} O — - ""‘Ir‘ e ‘ '7" r r”.-
.Ju...u-lv.b n 4‘. K4 ¢.L\~._I- afiA-*./.L H... b-1 VA.- 9.x; -l.__“.LlIJ US.- LL DJ. 5 ... ,_|.--1._.2-l ( .L. 2 -..U.

\ A ‘ —-° C ( '~~ -.-~ “A -' J“ 7-. ‘ '2 ' V‘ ", "0 ‘ ‘ ‘| v-1 ‘-
per alleraa) per gOuul Dulce aallj or 0:. Larger aAa/OI more

l
J.)

'0
._J
I

O
O

p , l} 1"» ~~ ~ ug} q -‘ .-~ 1 - .-_-L- l1"_ -.,
lre.lea~ ooses i:-’0;u034 an unneces, arl_d a- ulOOa concerorau_on

“"V -' " fl '1'“ . 2 ’3 J” - '2 '1’ '. n‘ ’L 11 L I "' ,~ 1 '
Suadli es ol one blooa eleuure laeLCaue aao bae Urea»-

,‘J. --r fi-a'.r""‘. ' ‘. V1 4.“... Y'Nfl‘ "'11 "3‘ J' ‘ "'0'? '\ M‘- .1 V" r“ ' H01 ' H *
eub Vs}..._£_...‘.u.'.. fl _Ll. Lain.) luau. CL, COLA—LIV O‘JULL..-1;.LM ll]. 3.“. mama»... Ma...)

/,~ -. 1. - .'.Lv, .zn ‘ n -- .- '-' j, -7.‘ *.-

JO,JOD; 11 one LadollJJ uaere was V'“f llUQle caarue. lacreased
’ooa“e* o“ e“l“e'ero~4va ”ww4mw n +nnon-finw nevew 310 1 mg a wnu?o_
-. yuuv.) .4. “.'.—..u. -L’ u....;.;..1-u “Lu—L _..-“b a. ugq. L4 ulna} .3 ‘J..U-LJ' \. by s.» ..i—v» ..
Y\('\ "‘z 31 1". 1“) V113. 7 a 1". —~ (1 T‘r‘."'-1c)w 3h ‘1”1 ‘vr 'L‘fi'fl "hp/‘1 I: 1 q: 3‘
‘Jvl‘ma .LA]. U$~b no:& Alba. Lil] 0U . __¢A Ji- LaJtJ~~OLo~ ...—“I U$$~J b... 11-1%.. If». L’O-
11'1‘6‘3 “l " 3 n l,""$‘w ‘3')!3“'. :- «v m *N‘a V"1)fi‘," p.11-on-wn l1
nu‘J. -— ‘.L 011—: Ob“ LJ()S ”Ll-1 d.v-- "05) 10L).‘_--v. '_._'_'_:.. 3J3.” J-Ovfi- «W5C v t'l';-"- 4 - H . ._._. \_L. -Qr‘u-_

fi ,

z ,1..'.. V: .'- ,. --- - -'n - 4.1 _-J., n I , a 4”? r' a“

zlne was aamlalseerel bJ hogan at one rage or ad Ussllns uaj.) LO.
a J 1 .1. ~ I -v 'Q -:

-’\ 1a: .1 4v ','-v . -‘ , p" n ‘ 2'7" V . " ‘
yer a-louraa) per yOflnd or ooa; JClUno

9 <~ q 4”v~fis 3w"’~\\W’ ‘ *'!+1 J” a 1 ~v~3w “»V‘mfl Jeww 1 era an .TW} n
.. _. e. U-..'. Lb Len”. ch; 4.0(1. u ell Ont. ...{ll “CAL LLU Lu), ULJ.\2 ‘09.; at- wee. u
I" 11 rue 0‘3 VI \"-= ”ia.-11 '1:\'3“|(30M‘: ‘A 1 7“: 07W 1 ant/ar.. fi.’\'fm~1h_‘. r1 ".“"‘ h '1’!‘ ’3“ L“ ‘.\
_. (_...4. .. Li‘s—L. n-1,..1.‘_'. ‘vu‘a l}. w LIV—oi. .’7-.»._‘..;.'.J. ....g...) v 9v V an. .1._'. .‘.‘U'J .3 x‘_4..'_ LIL/.1. U_4.L
..

a . _o o _t_ 4- o in _L1

,"","‘y“'f“}'l‘I/:~ '1 ‘ '1 '- mw ~
3.w.---—‘-.a.~ Va. UAOL- .'. VLI‘V K..- L o

RWM USE ONLY