C E R T A I N H E M A T O L O G I C A N D BLOOD C H E M I C A L VALUES

I N EEA.GLE DOGS

James F. V o n d r u s k a , D . V . M . a n d

R o n a l d 8. G r e c o , B . S . *

Summary

C e r t a i n h e m a t o l o g i c a n d b l o o d b i o c h e m i c a l p a r a m e t e r s w e r e

d e t e r m i n e d on o v e r 200 p u r e b r e d B e a g l e d o g s . Mean v a l u e s , s t a n d a r d

d e v i a t i o n s , a n d m i n i m a l - m a x i n a l v a l u e s w e r e d e t e r m i n e d f o r v a r i o u s

a g e c a t e g o r i e s a n d f o r e a c h s e x . The f o l l o w i n g p a r a n e t e r s w e r e

m e a s u r e d a n d compared t o c i . t e d v a l u e s w h e r e t h e y w e r e a v a i l a b i e :

h e m o g l o b i n c o n c e n t r a t i o n , p a c k e d c e l l v o l u m e , t o t a l e r y t h r o c y t e ,

t o t a l a n d d i f f e r e n t i a l l e u k o c y t e c o u n t s , p l a t e l e t a n d r e t i c u l o c y t e

c o u n t s , c e l l u l a r i n d i c e s , p r o t h r o m b i n t i m e , c o a g u l a t i o n t i m e ,

s e r u m c o n c e n t r a t i o n s o f g l u c o s e , u r e a n i t r o g e n , c r e a t i n i n e , b i l i -

r u b i n , u r i c a c i d , c h o l e s t e r o l , s o d i u m , p o t a s s i u m a n d c h l o r i d e , a n d

s e r u m enzyme a c t i v i t i e s o f a l k a l i n e p h o s p h a t a s e , g l u t a m i c p y r u v i c

t r a n s a m i n a s e , g l u t a m i c o x a l a c e t i c t r a n s a m i n a z e arid l a c t i c dehy-

d r o g e n a s e .

* D r . James F . V o n d r u s k a w a s s t a f f v e t e r i n a r i a n a n d s e n i o r g r o u p l e a d e r of p r i m a t e t o x i c o l o g y a n d c l i n i c a l p a t h o l o g y a n d M r . R o n a l d A . G r e c o i s g r o u p l e a d e r o f c l i n i c a l p a t h o l o g y a t I n d u s t r i a l BIO-TEST L a b o r a t o r i e s , I n c . , N o r t h b r o o k , I l l i n o i s .

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The Beagle dog is one of t h e most commonly used r e sea rch animals

i n t h e f i e l d o f b io logy . I n t h e a r e a of t ox ico logy , g r e a t emphasis i s

placed upon hematologic and blood biochemical para:.::e-rs i n a s s e s s i n g

t h e r e s u l t s of many t o x i c o l o g i c a l s t u d i e s . For t h i s purpose w e have

compiled a summary of normal hematologic and blood biochemical v a l u e s ,

ca t egor i zed by t h e sex and age of the s u b j e c t an imals . These animals

are cons idered t o be r e p r e s e n t a t i v e of our contro; Beagle dog popula t ion .

Materials and Methods

Y

The s u b j e c t an imals a r e purebred Beagle dogs, which a r e bred by

our l a b o r a t o r y f o r our e x c l u s i v e use i n tox ico logy i n v e s t i g a t i o n s : All

dogs had been immunized a g a i n s t can ine d i s t emper , i n f e c t i o u s canine

h e p a t i t i s and l e p t o s p i r o s i s a t t h e age of approximately t h r e e t o fou r

months, and r ece ived boos te r immunizations annua l ly t h e r e a f t e r . T h e dogs

were housed i n groups of fou r of each sex i n conc re t e f loo red combination

indoor-outdoor runs .

and d r i n k i n g w a t e r w a s a v a i l a b l e a t a l l t i m e s .

of each sex w a s used on t h i s s t u d y , though n o t a l l parameters were

1 They were maintained on a d i e t of Pur ina Dog Meal

A t o t a l of over 100 dogs

measured i n every animal , and n o t every animal w a s examined a t each t i m e

i n t e r v a l .

Blood w a s c o l l e c t e d by puncture of t h e e x t e r n a l j u g u l a r v e i n us ing

2 a 20 gauge, 1 inch need le and c o l l e c t i n g t h e blood i n Vacuta iners Blood

t o be used f o r performing t h e complete blood count w a s c o l l e c t e d i n a tube

con ta in ing EDTA and t h a t t o be used f o r prothrombin t i m e de t e rmina t ions

1 Ralston-Purina Company, S t . Louis , Missouri .

Vacutainer@, hecton-Dickinson & C o . , Ruther ford , N e w Jersey. 2

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w a s c o l l e c t e d i n a tube con ta in ing sodium o x a l a t e . Serum f o r blood b io-

chemical measurements w a s ob ta ined from blood c o l l e c t e d i n similiar tubes

con ta in ing no an t i coagu lan t and allowed t o c l o t i n an ice ba th . Micro-

s cop ic s l i d e s f o r d i f f e r e n t i a l l eukocyte coun t s were prepared from t h e

blood sample c o l l e c t e d i n EDTA and immediately f i x e d i n a b s o l u t e methanol.

A l l blood c o l l e c t i o n s were performed i n t h e morning a f t e r t he dogs had

been f a s t e d f o r 18 hours . .

T o t a l e r y t h r o c y t e and leukocyte coun t s , hemoglobin concen t r a t ion

and mean co reuscu la r volume were determined d i r e c t l y wi th an e l e c t r o n i c

1 ce l l coun te r . Hematocri t , mean corpuscular hemoglobin and mean corpuscu-

l a r hemoglobin concen t r a t ion were a l l c a l c u l a t e d v a l u e s which were

determined by t h i s same ins t rument .

D i f f e r e n t i a l l eukocyte counts were made from smears s t a i n e d wi th

One hundred ce l l s Wright-Giemsa s t a i n and examined under o i l immersion.

were counted from each s l i d e . P l a t e l e t e s t i m a t i o n s were a l s o made from

t h e s e s l i d e s . Re t i cu locy te counts w e r e made from s l i d e s s t a i n e d wi th

new methylene b l u e (1).

Blood coagu la t ion parameters were performed manually. Prothrombin

t i m e was done by t h e one-step method of Quick (2) and coagu la t ion t i m e by

t h e method of Schalm ( 3 ) . Blood biochemical ana lyses performed manually

were: t o t a l b i l i r u b i n , by t h e method of Malloy and Evelyn ( 4 ) ; - u r i c a c i d ,

by t h e method of Henry e t a l . (I); and c h o l e s t e r o l , by t h e method of Huang

e t a l . (6). --

Model S Coul te r Counter, Coul te r E l e c t r o n i c s , Hia leah , F lo r ida .

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Glucose and blood urea n i t rogen were performed with the a i d of an 1

automated continuous flow analyzing instrument by the simultaneous method

o f Mabry -- e t a l . (z), which w a s performed i n conjunction with the serum

a l k a l i n e phosphatase method of Babson (4) and Babson -- e t a l . (2) and the

serum g l u t d c pyruvic transaminase method of Levine and H i l l (u). Serum

glutamic o x a l a c e t i c transaminase w a s performed as an ind iv idua l opera t ion ,

a l s o by the method of Levine and H i l l . L a c t i c dehydrogenase w a s determined

by an automated method of Levy 7 - e t al. (11) and c r e a t i n i n e by the method of

Hawk e t a l . (121, which w a s modified for automation. Automated flame photo-

metry w a s usgd f o r t h e determination of sodium and potassium, which w a s

performed s imultaneously with t h e co lo r ime t r i c procedure f o r ch lo r ide by

Zal l e t a l . (Q).

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Results and Discussion

The r e s u l t s of the hemglobin concent ra t ion , packed ce l l volume ,

t o t a l e ry th rocy te count, and corpuscular constant determinat ions are presented

i n Tables 1 and 2. The r e s u l t s have been grouped by t h e age of the dogs,

and mean va lues , s tandard devia t ions , and ranges were computed sepa ra t e ly

f o r males and females.

Because of d i f f e rences i n age and source o f Beagles and methodology,

our results would not be expected t o be exac t ly comparable t o many of those

i n the l i t e r a t u r e . The hemglobin values obtained i n t h i s compilation of

d a t a i n d i c a t e t h a t t h i s value normally increases t o i ts maximum a t about the

t i m e of sexual matur i ty (about 8 months of age) . This observat ion has been

A u t o h a l y z e r , Technicon Instruments , Tarrytown, New York.

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corroborated (3, as have the hemoglobin values f o r a d u l t dogs (14, - 15, - - - - 16, 17 , 18 , 19 , 2, 21). lower than those prev ious ly repor ted (14, IS, 16, ' 7 , - I - 18) b u t a r e equi-

v a l e n t t o o t h e r s (3, 19, 20, 3). t o methodology, s i n c e many of the c i t e d values were determined by cen-

Hematocrit values i n mature dogs are genera l ly

This d i f f e r e n c e i s very l i k e l y due

t r i f u g a t i o n r a t h e r than ca l cu la t ed as i n our da ta . I n t h i s parameter an

inc reas ing t rend is a l s o noted up t o tile time of n a t u r i t y . Erythrocyte

counts tend t o inc rease wi th matur i ty , and are h igher than some of the

c i t e d l i t e r a t u r e (17, 18, l9, 2) and approximately equiva len t t o o the r

d a t a (2, 16). w

Mean corpuscular volume va lues are lower than most of t h e c i t e d

va lues , (3, - - 1 7 , 1 8 ) bu t are equiva len t i n some cases (19, - 2). Much of the

d i f f e rence is a func t ion of t he t rend f o r t he c i t e d hematocri t values t o be

higher . The c i t e d values f o r mean corpuscular hemoglobin concent ra t ion are

c o n s i s t e n t l y lower than our values (3, 17, 18, 2, s), whereas the va lues

f o r mean corpuscular hemoglobin e i t h e r agree (3, 2, - 20) o r are higher than

our values (17, 18). No t rends due t o inc reas ing age are seen among the

corpuscular cons tan ts i n our d a t a o t h e r than a s l i g h t i nc rease i n mean

corpuscular hemoglobin concent ra t ion which para l le l s the inc reas ing hemo-

g lobin concent ra t ion .

Our p l a t e l e t es t imat ions are much higher than the va lue reported

by one au thor (17) - bu t f i t i n t o the broad range c i t e d by another (3). Considerable v a r i a t i o n w a s seen i n the r e t i c u l o c y t e counts i n our d a t a and

i n the l i t e r a t u r e (3, l4, 17, 2, 2).

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The r e s u l t s of the t o t a l and d i f f e r e n t i a l l eukocyte counts are

p resen ted i n Tables 3 and 4 . This data was a l s o grouped by t h e age of

t h e an imals , and mean v a l u e s , s t a n d a r d d e v i a t i o n s and ranges were com-

puted s e p a r a t e l y f o r males and females. The t o t a l counts tend t o be

s l i g h t l y h ighe r than i n the c i ted l i t e r a t u r e and the d i f f e r e n c e s appear

t o be due p r i m a r i l y t o n e u t r o p h i l i a (3, 14, 15, 16, 17, 19, 22, 2). No c o n s i s t e n t t r e n d w a s noted wi th i n c r e a s i n g age.

The blood biochemical d a t a are p resen ted i n Tables 5 t o 3 and

are grouped 4n a manner s i m i l a r t o t h a t used f o r the hematoloRic da t a .

Serum glucose va lues remain r e l a t i v e l y c o n s t a n t w i th age and are comparable

wi th va lues r epor t ed from one source (22) b u t cons ide rab ly lower than

blood g lucose va lues from ano the r sou rce (19) . - Rlood Elucose va lues can-

n o t b e compared t o serum g lucose , and serum p,lucose d i f f e r e n c e s may be

due t o methodology. Blood u rea n i t r o g e n va lues are n o t a f f e c t e d by axe

and compare favorably wi th va lues from one source (14) b u t are gene ra l ly

s l i g h t l y lower than most r epor t ed va lues (17, 2, 22).

could be l o c a t e d f o r serum a l k a l i n e phosphatase , serum glu tamic pyruvic

t r a n s aminase o r se rum g l u t amic oxa l ace t i c t r a n s aminas e whi ch were rep o r t ed

i n t h e same u n i t s as ou r da t a . A s t eady decrease was noted i n the serum

a l k a l i n e phosphatase va lues w i t h age , bu t t he t ransaminase va lues d i d not

No l i t e r a t u r e va lues

f l u c t u a t e i n any manner which could be i n t e r p r e t e d as a t r end .

No prothrombin t i m e o r coagu la t ion t i m e va lues were a v a i l a b l e f o r

comparison. L a c t i c dehydrogenase va lues r epor t ed i n the same u n i t s were

l i k e w i s e lack ing . No p a r t i c u l a r t r end was seen w i t h any of t h e s e parameters .

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C r e a t i n i n e v a l u e s w e r e lower t h a n t w o cases r e p o r t e d i n t h e l i t e r a t u r e

(17 , - - 19) b u t approximate ly e q u a l t o a t h i r d ( 2 2 ) . - B i l i r u b i n v a l u e s d i d

n o t d i f f e r from t h e c i t e d v a l u e s (17, - 22) nor d i d c h o l e s t e r o l (19 , - 22). Uric a c i d w a s n o t e d t o i n c r e a s e s l i g h t l y between 5 and 6% months of a g e ,

t h e o n l y i n t e r v a l s a t which t h i s p a r a m e t e r w a s measured.

tended t o b e h i g h e r t h a n t h e l i t e r a t u r e v a l u e s (l7, 2, 22).

Thse v a l u e s

Serum sodium compared f a v o r a b l y w i t h t h e c i t e d v a l u e s i n a l l

cases (2, 2, 2, 21) e x c e p t one which was c o n s i d e r a b l y h i g h e r (22).

P o t a s s i u m v a u e s were e q u i v a l e n t t o t h e r e p o r t e d v a l u e s (2, 17, 19, 2, 22) as w e r e t h e c h l o r i d e v a l u e s (14, - - . - 1 7 , 19, - 22). -

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